SMART MODE:

NORMAL GENOMIC

• Plotnikov AN et al.
• Crystal structure of fibroblast growth factor 9 reveals regions implicated in dimerization and autoinhibition.
• J Biol Chem. 2001; 276: 4322-9
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• Fibroblast growth factors (FGFs) constitute a large family of heparin-binding growth factors with diverse biological activities. FGF9 was originally described as glia-activating factor and is expressed in the nervous system as a potent mitogen for glia cells. Unlike most FGFs, FGF9 forms dimers in solution with a K(d) of 680 nm. To elucidate the molecular mechanism of FGF9 dimerization, the crystal structure of FGF9 was determined at 2.2 A resolution. FGF9 adopts a beta-trefoil fold similar to other FGFs. However, unlike other FGFs, the N- and C-terminal regions outside the beta-trefoil core in FGF9 are ordered and involved in the formation of a 2-fold crystallographic dimer. A significant surface area (>2000 A(2)) is buried in the dimer interface that occludes a major receptor binding site of FGF9. Thus, we propose an autoinhibitory mechanism for FGF9 that is dependent on sequences outside of the beta-trefoil core. Moreover, a model is presented providing a molecular basis for the preferential affinity of FGF9 toward FGFR3.

• Blair SS
• Wnts, signaling and sulfates.
• Sci STKE. 2001; 2001: 32-32
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• Questions remain about the signaling pathways that control pattern formation during development. Blair describes how sulfated glycosaminoglycans affect several developmentally important signaling pathways, including Wnt-Wingless, Fibroblast growth factor, Hedgehog, and Bone morphogenetic protein-4 signaling. A new secreted sulfatase, Qsulf1, regulates the sensitivity of vertebrate cells to Wnts, possibly by modifying the sulfation of glycosaminoglycans.

• Kurban G et al.
• [Expression and intracytoplasmic signal transduction pathway of fibroblast growth factor (FGF)-10 in human cervical cancer cell lines]
• J Nippon Med Sch. 2001; 68: 253-8
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• Fibroblast growth factor (FGF) -10 is a new member of the FGF family initially reported in Japan. It is mainly synthesized by mesenchymal cells and acts on epithelial cells in a paracrine manner. FGF-10 actions are dependent on their binding to the iiib form of FGF receptor 2 (FGFR2) iiib, also known as keratinocyte growth factor receptor (KGFR). FGF-10 has high amino acid homology to keratinocyte growth factor (KGF) and plays an important role in fetal limb and lung development and skin wound healing. In the present study, the expression of FGF-10 and FGFR2 iiib messenger RNA (mRNA) in two different human uterine cervical cancer cell lines (CaSki and ME-180) were examined. Both CaSki and ME-180 cells expressed FGFR2 iiib mRNA, while only CaSki cells expressed FGF-10 mRNA and protein. Recombinant FGF-10 (1 ng/ml) increased the growth rate of ME-180 cells and also enhanced mitogen-activated protein kinase (MAPK) phosphorylation of the cells. These data indicate that FGF-10 may directly promote the growth of squamous cell cancer in the uterine cervix via the MAPK pathway.

• So F, Daley TD, Jackson L, Wysocki GP
• Immunohistochemical localization of fibroblast growth factors FGF-1 and FGF-2, and receptors FGFR2 and FGFR3 in the epithelium of human odontogenic cysts and tumors.
• J Oral Pathol Med. 2001; 30: 428-33
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• Acidic (FGF-1) and basic (FGF-2) fibroblast growth factors are members of a family of growth factors that function in growth, differentiation and regeneration of a variety of tissues. Their presence in human odontogenic cysts and tumors has not been previously investigated. This study was designed to detect immunohistochemically the presence of these factors and two fibroblast growth factor receptors (FGFR2 and FGFR3) in a cross section of odontogenic cysts and tumors, to determine if they may be involved in the differentiation of odontogenic epithelium or, more specifically, in the development of particular cysts or tumors. Archival formalin-fixed paraffin-embedded tissues were used. With some exceptions, FGF-2 and the receptor FGFR2, were found in the cytoplasm and occasionally in the nuclei of cells of odontogenic epithelium, while FGF-1 and the receptor FGFR3, were absent or only focally or weakly detected, using standard immunohistochemical techniques. The data are similar to those published for normal murine odontogenesis, suggesting that these factors are associated with odontogenic differentiation rather than pathogenesis. The presence of significant nuclear staining in odontogenic epithelium associated with embryonic mesenchyme in ameloblastic fibromas and ameloblastic fibro-odontomas suggests that FGF-2 may be involved in directing nuclear activity at the histodifferentiation stage of odontogenesis.

• Kim I, Moon S, Yu K, Kim U, Koh GY
• A novel fibroblast growth factor receptor-5 preferentially expressed in the pancreas(1).
• Biochim Biophys Acta. 2001; 1518: 152-6
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• Using the polymerase chain reaction on human embryonic cDNAs, we isolated a cDNA encoding a novel 504 amino acid protein, termed fibroblast growth factor receptor (FGFR)-5, which is highly homologous to known FGFRs. The NH(2)-terminal portion of FGFR5 contains a putative secretory signal sequence, three typical immunoglobulin-like domains, six cysteines, and an acidic box, but no HAV motif. The COOH-terminal portion of FGFR5 contains one transmembrane domain but no intracellular kinase domain. Recombinant FGFR5 expressed in COS-7 cells is not secreted, but recombinant truncated FGFR5 lacking the predicted transmembrane domain is secreted. Acidic fibroblast growth factor (aFGF) and basic fibroblast growth factor (bFGF) do not bind to FGFR5. Among 23 adult human tissues, FGFR5 mRNA is preferentially expressed in the pancreas. These results suggest that FGFR5 may provide a binding site for some other fibroblast growth factors and may regulate some pancreatic function.

• Cailliau K, Browaeys-Poly E, Vilain JP
• Fibroblast growth factors 1 and 2 differently activate MAP kinase in Xenopus oocytes expressing fibroblast growth factor receptors 1 and 4.
• Biochim Biophys Acta. 2001; 1538: 228-33
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• The mitogen-activated protein kinase (MAP kinase) signalling cascade activated by fibroblast growth factors (FGF1 and FGF2) was analysed in a model system, Xenopus oocytes, expressing fibroblast growth factor receptors (FGFR1 and FGFR4). Stimulation of FGFR1 by FGF1 or FGF2 and FGFR4 by FGF1 induced a sustained phosphorylation of extracellular signal-regulated protein kinase 2 (ERK2) and meiosis reinitiation. In contrast, FGFR4 stimulation by FGF2 induced an early transient activation of ERK2 and no meiosis reinitiation. FGFR4 transduction cascades were differently activated by FGF1 and FGF2. Early phosphorylation of ERK2 was blocked by the dominant negative form of growth factor-bound protein 2 (Grb2) and Ras, for FGF1-FGFR4 and FGF2-FGFR4. The phosphatidylinositol 3-kinase (PI3 kinase) inhibitors wortmannin and LY294002 only prevented the early ERK2 phosphorylation triggered by FGF2-FGFR4 but not by FGF1-FGFR4. ERK2 phosphorylation triggered by FGFR4 depended on the Grb2/Ras pathway and also involved PI3 kinase in a time-dependent manner.

• Lozano RM et al.
• Solution structure and interaction with basic and acidic fibroblast growth factor of a 3-kDa human platelet factor-4 fragment with antiangiogenic activity.
• J Biol Chem. 2001; 276: 35723-34
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• Platelet factor-4 is a protein belonging to the family of ELR-negative CXC chemokines which binds to fibroblast growth factor and inhibits its mitogenic activity. Platelet factor-4 also inhibits tumor growth by mechanisms involving antiangiogenesis. Antiangiogenic activity in vitro has also been shown for the 24-residue C-terminal fragment of the protein, which decreases the affinity between basic fibroblast growth factor and its cell-surface receptor. In this study, the preferential conformation of this fragment in solution has been determined and has been found to be composed of two helical subdomains. In addition, we show that the fragment forms a specific 1:1 complex with acidic and basic fibroblast growth factors and that both subdomains are probably required for inhibition of fibroblast growth factor-driven mitogenesis. Finally, we show that the binding of the fragment alters the structure of the fibroblast growth factors, although some of such alterations do not seem related with the inhibition of mitogenic activity. Since this fragment has recently been shown to inhibit fibroblast growth factor-induced angiogenesis in vivo when injected intraperitoneally, these results are relevant for developing new antiangiogenic treatments.

• Spencer-Dene B, Dillon C, Fantl V, Kerr K, Petiot A, Dickson C
• Fibroblast growth factor signalling in mouse mammary gland development.
• Endocr Relat Cancer. 2001; 8: 211-7
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• Fibroblast growth factors (Fgfs) and their receptors are important intercellular signalling molecules involved in many aspects of animal development. The aberrant expression of the Fgfs or the inappropriate activation of their cell surface receptors have been implicated in tumorigenesis. Here, we describe the evidence that as well as playing a critical role in the formation of the mammary primordia during embryogenesis, signalling by Fgfs is necessary for optimal lobuloalveolar development of the mouse mammary gland during pregnancy.

• Rhinn M, Brand M
• The midbrain--hindbrain boundary organizer.
• Curr Opin Neurobiol. 2001; 11: 34-42
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• Cell fate in the cephalic neural primordium is controlled by an organizer located at the midbrain-hindbrain boundary. Studies in chick, mouse and zebrafish converge to show that mutually repressive interactions between homeodomain transcription factors of the Otx and Gbx class position this organizer in the neural primordium. Once positioned, independent signaling pathways converge in their activity to drive organizer function. Fibroblast growth factors secreted from the organizer are necessary for, and sufficient to mimic, organizer activity in patterning the midbrain and anterior hindbrain, and are tightly controlled by feedback inhibition.

• Tuominen H et al.
• Expression and glycosylation studies of human FGF receptor 4.
• Protein Expr Purif. 2001; 21: 275-85
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• Fibroblast growth factor receptor subtype 4 (FGFR4) has been shown to have special activation properties and just one splicing form, unlike the other FGFRs. FGFR4 overexpression is correlated with breast cancer and therefore FGFR4 is a target for drug design. Our aim is to overexpress high amounts of homogeneous FGFR4 extracellular domain (FGFR4(ed)) for structural studies. We show that baculovirus-insect cell-expressed FGFR4(ed) is glycosylated on three (N88, N234, and N266) of the six possible N-glycosylation sites but is not O-glycosylated. The deglycosylated triple mutant was expressed and had binding properties similar to those of glycosylated FGFR4(ed), but was still heterogeneous. Large amounts of FGFR4(ed) have been produced into inclusion bodies in Escherichia coli and refolded at least partly correctly but the refolded E. coli-produced FGFR4(ed) still aggregates. Copyright 2001 Academic Press.

• Alitalo K
• The fatal detachment.
• Nat Cell Biol. 2001; 3: 1579-1579

• Mathis L, Kulesa PM, Fraser SE
• FGF receptor signalling is required to maintain neural progenitors during Hensen's node progression.
• Nat Cell Biol. 2001; 3: 559-66
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• Previous analyses of labelled clones of cells within the developing nervous system of the mouse have indicated that descendants are initially dispersed rostrocaudally followed by more local proliferation, which is consistent with the progressing node's contributing descendants from a resident population of progenitor cells as it advances caudally. Here we electroporated an expression vector encoding green fluorescent protein into the chicken embryo near Hensen's node to test and confirm the pattern inferred in the mouse. This provides a model in which a proliferative stem zone is maintained in the node by a localized signal; those cells that are displaced out of the stem zone go on to contribute to the growing axis. To test whether fibroblast growth factor (FGF) signalling could be involved in the maintenance of the stem zone, we co-electroporated a dominant-negative FGF receptor with a lineage marker, and found that it markedly alters the elongation of the spinal cord primordium. The results indicate that FGF receptor signalling promotes the continuous development of the posterior nervous system by maintaining presumptive neural progenitors in the region near Hensen's node. This offers a potential explanation for the mixed findings on FGF in the growth and patterning of the embryonic axis.

• Uematsu F, Jang JH, Kan M, Wang F, Luo Y, McKeehan WL
• Evidence that the intracellular domain of FGF receptor 2IIIb affects contact of the ectodomain with two FGF7 ligands.
• Biochem Biophys Res Commun. 2001; 283: 791-7
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• Models of the oligomeric FGF signaling complex, including those derived from crystal structures, vary in stoichiometry and arrangement of the three subunits comprised of heparin/heparan sulfate chains, FGFR tyrosine kinase and activating FGF. Here, using covalent affinity crosslinking of radiolabeled FGF7 to binary complexes of FGFR2IIIb and heparin, we show that two molecules of FGF7 contact each FGFR2IIIb. This supports models that propose a dimeric complex of two units with stoichiometry 1 FGF:1 FGFR in which each FGF contacts both FGFR. The bivalent FGF7 contact was dependent on the full-length amino terminus of FGF7alpha and the intracellular domain of FGFR2IIIb extending through the juxtamembrane domain and the beta1 and beta2 strands of the kinase which is required for ATP binding. We propose that the differences in crosslinking report differences in relationships among subunits in the ectodomain of the complex that are affected by the amino terminus of FGF and the FGFR intracellular domain. From this, we suggest the corollary that conformational relationships among subunits in the ectodomain are transmitted to the intracellular and ATP binding domains during activation of the complex. Copyright 2001 Academic Press.

• Ford-Perriss M, Abud H, Murphy M
• Fibroblast growth factors in the developing central nervous system.
• Clin Exp Pharmacol Physiol. 2001; 28: 493-503
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• 1. It is now clear that members of the fibroblast growth factor (FGF) family have multiple roles during the formation of the central nervous system (CNS). 2. There are at least 23 members of the FGF family and, of these, 10 are expressed in the developing CNS, along with four FGF receptors (FGFR-1-4). 3. The present review discusses the roles of these FGFs, with emphasis on FGF-2, FGF-8, FGF-15 and FGF-17. Fibroblast growth factors-2 and -15 are generally expressed throughout the developing CNS, whereas FGF-8 and FGF-17 are tightly localized to specific regions of the developing brain and are only expressed in the embryo during the early phases of proliferation and neurogenesis. 4. Expression studies on FGFRs in the chick and mouse indicate that FGFR-1 is most generally expressed, whereas FGFR-2 and FGFR-3 show highly localized but changing patterns of expression throughout CNS development. The FGFR-4 has been localized to the developing CNS in fish but not at a detailed level, as yet, in chick or mouse. 5. A picture is emerging from these studies that particular FGFs signal through specific receptors in a highly localized manner to regulate the development of different regions of the brain. 6. This picture has been demonstrated so far for the developing cortex (FGF-2-/- mice), the forebrain and midbrain (FGF-8 hypomorphs) and the cerebellum (FGF-17/FGF-8 mutant mice). In addition, generation of mutant animals deleted for FGFR-1 and FGFR-2b IIIb demonstrate their importance in FGF signalling. 7. However, there are significant gaps in our knowledge of the localization of members of the FGF family and their receptors. More detailed information on the spatio-temporal mapping of FGFs and FGFR isoforms is required in order to understand the molecular mechanisms through which FGFs signal.

• Revest JM, Spencer-Dene B, Kerr K, De Moerlooze L, Rosewell I, Dickson C
• Fibroblast growth factor receptor 2-IIIb acts upstream of Shh and Fgf4 and is required for limb bud maintenance but not for the induction of Fgf8, Fgf10, Msx1, or Bmp4.
• Dev Biol. 2001; 231: 47-62
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• Mice deficient for FgfR2-IIIb were generated by placing translational stop codons and an IRES-LacZ cassette into exon IIIb of FgfR2. Expression of the alternatively spliced receptor isoform, FgfR2-IIIc, was not affected in mice deficient for the IIIb isoform. FgfR2-IIIb(-/-) (lac)(Z) mice survive to term but show dysgenesis of the kidneys, salivary glands, adrenal glands, thymus, pancreas, skin, otic vesicles, glandular stomach, and hair follicles, and agenesis of the lungs, anterior pituitary, thyroid, teeth, and limbs. Detailed analysis of limb development revealed an essential role for FgfR2-IIIb in maintaining the AER. Its absence did not prevent expression of Fgf8, Fgf10, Bmp4, and Msx1, but did prevent induction of Shh and Fgf4, indicating that they are downstream targets of FgfR2-IIIb activation. In the absence of FgfR2-IIIb, extensive apoptosis of the limb bud ectoderm and mesenchyme occurs between E10 and E10.5, providing evidence that Fgfs act primarily as survival factors. We propose that FgfR2-IIIb is not required for limb bud initiation, but is essential for its maintenance and growth.

• Vasiliauskas D, Stern CD
• Patterning the embryonic axis: FGF signaling and how vertebrate embryos measure time.
• Cell. 2001; 106: 133-6

• Jackson TA, Schweppe RE, Koterwas DM, Bradford AP
• Fibroblast growth factor activation of the rat prl promoter is mediated by pkcdelta.
• Mol Endocrinol. 2001; 15: 1517-28
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• Fibroblast growth factors play a critical role in cell growth, development, and differentiation and are also implicated in the formation and progression of tumors in a variety of tissues including pituitary. We have previously shown that fibroblast growth factor activation of the rat PRL promoter in GH4T2 pituitary tumor cells is mediated via MAP kinase in a Ras/Raf-1-independent manner. Herein we show using biochemical, molecular, and pharmacological approaches that PKCdelta is a critical component of the fibroblast growth factor signaling pathway. PKC inhibitors, or down-regulation of PKC, rendered the rat PRL promoter refractory to subsequent stimulation by fibroblast growth factors, implying a role for PKC in fibroblast growth factor signal transduction. FGFs caused specific translocation of PKCdelta from cytosolic to membrane fractions, consistent with enzyme activation. In contrast, other PKCs expressed in GH4T2 cells (alpha, betaI, betaII, and epsilon) did not translocate in response to fibroblast growth factors. The PKCdelta subtype-selective inhibitor, rottlerin, or expression of a dominant negative PKCdelta adenoviral construct also blocked fibroblast growth factor induction of rat PRL promoter activity, confirming a role for the novel PKCdelta isoform. PKC inhibitors selective for the conventional alpha and beta isoforms or dominant negative PKCalpha adenoviral expression constructs had no effect. Induction of the endogenous PRL gene was also blocked by adenoviral dominant negative PKCdelta expression but not by an analogous dominant negative PKCalpha construct. Finally, rottlerin significantly attenuated FGF-induced MAP kinase phosphorylation. Together, these results indicate that MAP kinase-dependent fibroblast growth factor stimulation of the rat PRL promoter in pituitary cells is mediated by PKCdelta.

• Pickles JO
• The expression of fibroblast growth factors and their receptors in the embryonic and neonatal mouse inner ear.
• Hear Res. 2001; 155: 54-62
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• Four different fibroblast growth factor receptors (FGFR) are known, three of which have splice variants (known as the b and c variants) in the FGF-binding domain, to give different patterns of sensitivity to the different FGFs. The expression of the b and c variants of the FGF receptors, together with the expression of the ligands FGF1, FGF2, FGF3, FGF7, FGF8b and FGF8c, was determined by quantitative reverse transcription-polymerase chain reaction in developing whole mouse inner ears, and in dissected components of the postnatal mouse inner ear. At embryonic age (E)10.5 days, when the otocyst is a simple closed sac, the receptor most heavily expressed was FGFR2b, relative to the postnatal day 0 level. Over the period E10.5-E12.5, during which the structures of the inner ear start to form, the expression of the different FGF receptors increased 10(2)-10(4) fold per unit of tissue, and there was a gradual switch towards expression of the 'c' splice variants of FGFR2 and FGFR3 rather than the 'b' variants. At E10.5, the ligands most heavily expressed, relative to the postnatal day 0 level, were FGF3, FGF8b and FGF8c. In the postnatal inner ear, the patterns of expression of receptors and ligands tended to be correlated, such that receptor variants were expressed in the same regions as the ligands that are known to activate them effectively. The neural/sensory region expressed high levels of FGFR3c, and high levels of the ligand FGF8b. The same area also expressed high levels of FGFR1b and FGFR2b, and high levels of FGF3. The lateral wall of the cochlea (including the stria vascularis and the spiral ligament) expressed high levels of FGFR1c and FGF2. It is suggested that the different FGF receptors and ligands are expressed in a spatially coordinated pattern, to selectively program cochlear development.

• Greenwald JA et al.
• In vivo modulation of FGF biological activity alters cranial suture fate.
• Am J Pathol. 2001; 158: 441-52
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• Gain-of-function mutations in fibroblast growth factor receptors have been identified in numerous syndromes associated with premature cranial suture fusion. Murine models in which the posterior frontal suture undergoes programmed fusion after birth while all other sutures remain patent provide an ideal model to study the biomolecular mechanisms that govern cranial suture fusion. Using adenoviral vectors and targeted in utero injections in rats, we demonstrate that physiological posterior frontal suture fusion is inhibited using a dominant-negative fibroblast growth factor receptor-1 construct, whereas the normally patent coronal suture fuses when infected with a construct that increases basic fibroblast growth factor biological activity. Our data may facilitate the development of novel, less invasive treatment options for children with craniosynostosis.

• Hensel F et al.
• A novel proliferation-associated variant of CFR-1 defined by a human monoclonal antibody.
• Lab Invest. 2001; 81: 1097-108
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• The germline coded human monoclonal IgM antibody 103/51 was isolated from a gastric carcinoma patient. This antibody binds to a 130-kd membrane molecule and has a mitotic effect on tumor cells in vitro. To characterize the target, we sequenced the protein and showed that the antibody binds to the cysteine-rich fibroblast growth factor receptor (CFR)-1, which is highly homologous to MG-160 and the E-selectin-ligand (ESL)-1. The epitope was determined by glycosidase-digestion experiments to be an N-linked carbohydrate side chain. Immunohistochemistry was used to investigate the tissue distribution of CFR-1. Different healthy tissues were tested and only the collecting tubes of the kidney, the Golgi apparatus, and the glomerular and fascicular zones of the adrenal gland stained positive. However, on malignant tissue the receptor is overexpressed in nearly all tested stomach cancers (12 of 15) and other tested carcinomas (13 of 15). Most interestingly, the receptor is also present in Helicobacter pylori gastritis and gastric dysplasia, but absent on uninflamed stomach mucosa. This restricted tissue pattern indicates that antibody 103/51 reacts with a membrane-bound variant of CFR-1, which is mainly expressed on transformed cells and precursor lesions and is essential for proliferation processes. The possible activity of antibody 103/51 as an activating ligand in these proliferative changes of gastric epithelial mucosa is discussed.

• Colvin JS, White AC, Pratt SJ, Ornitz DM
• Lung hypoplasia and neonatal death in Fgf9-null mice identify this gene as an essential regulator of lung mesenchyme.
• Development. 2001; 128: 2095-106
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• Mammalian lung develops as an evagination of ventral gut endoderm into the underlying mesenchyme. Iterative epithelial branching, regulated by the surrounding mesenchyme, generates an elaborate network of airways from the initial lung bud. Fibroblast growth factors (FGFs) often mediate epithelial-mesenchymal interactions and mesenchymal Fgf10 is essential for epithelial branching in the developing lung. However, no FGF has been shown to regulate lung mesenchyme. In embryonic lung, Fgf9 is detected in airway epithelium and visceral pleura at E10.5, but is restricted to the pleura by E12.5. We report that mice homozygous for a targeted disruption of Fgf9 exhibit lung hypoplasia and early postnatal death. Fgf9(-/-) lungs exhibit reduced mesenchyme and decreased branching of airways, but show significant distal airspace formation and pneumocyte differentiation. Our results suggest that Fgf9 affects lung size by stimulating mesenchymal proliferation. The reduction in the amount of mesenchyme in Fgf9(-/-) lungs limits expression of mesenchymal Fgf10. We suggest a model whereby FGF9 signaling from the epithelium and reciprocal FGF10 signaling from the mesenchyme coordinately regulate epithelial airway branching and organ size during lung embryogenesis.

• Furthauer M, Reifers F, Brand M, Thisse B, Thisse C
• sprouty4 acts in vivo as a feedback-induced antagonist of FGF signaling in zebrafish.
• Development. 2001; 128: 2175-86
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• In looking for novel factors involved in the regulation of the fibroblast growth factor (FGF) signaling pathway, we have isolated a zebrafish sprouty4 gene, based on its extensive similarities with the expression patterns of both fgf8 and fgf3. Through gain- and loss-of-function experiments, we demonstrate that Fgf8 and Fgf3 act in vivo to induce the expression of Spry4, which in turn can inhibit activity of these growth factors. When overexpressed at low doses, Spry4 induces loss of cerebellum and reduction in size of the otic vesicle, thereby mimicking the fgf8/acerebellar mutant phenotype. Injections of high doses of Spry4 cause ventralization of the embryo, an opposite phenotype to the dorsalisation induced by overexpression of Fgf8 or Fgf3. Conversely we have shown that inhibition of Spry4 function through injection of antisense morpholino oligonucleotide leads to a weak dorsalization of the embryo, the phenotype expected for an upregulation of Fgf8 or Fgf3 signaling pathway. Finally, we show that Spry4 interferes with FGF signaling downstream of the FGF receptor 1 (FGFR1). In addition, our analysis reveals that signaling through FGFR1/Ras/mitogen-activated protein kinase pathway is involved, not in mesoderm induction, but in the control of the dorsoventral patterning via the regulation of bone morphogenetic protein (BMP) expression.

• Christensen RN, Weinstein M, Tassava RA
• Fibroblast growth factors in regenerating limbs of Ambystoma: Cloning and semi-quantitative RT-PCR expression studies.
• J Exp Zool. 2001; 290: 529-40
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• Urodele amphibians (newts and salamanders) have the ability to regenerate amputated limbs throughout their life span. Because fibroblast growth factors (Fgfs) play important roles in developing limbs, we initiated studies to investigate these growth factors in regenerating limbs. Partial cDNAs of Fgf4, 8, and 10 were cloned from both the Mexican axolotl, Ambystoma mexicanum, and locally collected spotted salamander, Ambystoma maculatum, two salamanders well recognized for their regenerative capabilities. cDNAs from the two Ambystoma species were virtually identical, ranging from 97-100% nucleotide identity. Axolotl Fgf4, 8, and 10 showed nucleotide sequence identity with chick Fgf4, 8, and 10 of 79%, 83%, and 72%, respectively. RT-PCR showed that these growth factors are expressed in regenerating axolotl limbs as well as in developing salamander larvae at the three-digit forelimb stage. Fgf8 and 10 are upregulated during regeneration and thus may be involved in distal signaling similar to that of the developing chick limb. Fgf4, however, was undetectable by RT-PCR in the distal tips of regenerates, suggesting that it does not play the same role in limb regeneration that it does in limb development. We also investigated the role these Fgfs may have in the nerve-dependence of regeneration. They were expressed similarly in aneurogenic and innervated limbs, suggesting that they are not the neurotrophic factors responsible for nerve-dependence. Denervation prevented Fgf8 and 10 upregulation, suggesting Fgf pathways are downstream of nerve-dependence. These data highlight important similarities and differences in Fgf expression between limb development and limb regeneration. J. Exp. Zool. 290:529-540, 2001. Copyright 2001 Wiley-Liss, Inc.

• Krull CE, Krumlauf R
• Building from the bottom up.
• Nat Cell Biol. 2001; 3: 1389-1389

• Yu K, Ornitz DM
• Uncoupling fibroblast growth factor receptor 2 ligand binding specificity leads to Apert syndrome-like phenotypes.
• Proc Natl Acad Sci U S A. 2001; 98: 3641-3

• Sato T, Araki I, Nakamura H
• Inductive signal and tissue responsiveness defining the tectum and the cerebellum.
• Development. 2001; 128: 2461-9
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• The mes/metencephalic boundary (isthmus) has an organizing activity for mesencephalon and metencephalon. The candidate signaling molecule is Fgf8 whose mRNA is localized in the region where the cerebellum differentiates. Responding to this signal, the cerebellum differentiates in the metencephalon and the tectum differentiates in the mesencephalon. Based on the assumption that strong Fgf8 signal induces the cerebellum and that the Fgf8b signal is stronger than that of Fgf8a, we carried out experiments to misexpress Fgf8b and Fgf8a in chick embryos. Fgf8a did not affect the expression pattern of Otx2, Gbx2 or Irx2. En2 expression was upregulated in the mesencephalon and in the diencephalon by Fgf8a. Consequently, Fgf8a misexpression resulted in the transformation of the presumptive diencephalon to the fate of the mesencephalon. In contrast, Fgf8b repressed Otx2 expression, but upregulated Gbx2 and Irx2 expression in the mesencephalon. As a result, Fgf8b completely changed the fate of the mesencephalic alar plate to cerebellum. Quantitative analysis showed that Fgf8b signal is 100 times stronger than Fgf8a signal. Co-transfection of Fgf8b with Otx2 indicates that Otx2 is a key molecule in mesencephalic generation. We have shown by RT-PCR that both Fgf8a and Fgf8b are expressed, Fgf8b expression prevailing in the isthmic region. The results all support our working hypothesis that the strong Fgf8 signal induces the neural tissue around the isthmus to differentiate into the cerebellum.

• Weinstein DC, Hemmati-Brivanlou AA
• Src family kinase function during early Xenopus development.
• Dev Dyn. 2001; 220: 163-8
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• The Src family kinase Laloo functions downstream of the fibroblast growth factor (FGF) receptor, as a component of a signaling pathway required for mesoderm formation. In this study, we first show that Laloo expression can mimic FGF function in a distinct context, suggesting that the Src-related kinases are general transducers of FGF signaling during development. We then characterize the role of the SH-PTP2 phosphatase, and of bone morphogenetic protein (BMP)-pathway signaling, in mesoderm induction by Laloo. Finally, we demonstrate that the mesoderm-inducing activity of the Src family kinases is not limited to Laloo, indicating a degree of functional overlap between members of this gene family during early vertebrate development. Copyright 2001 Wiley-Liss, Inc.

• Hertz JM, Juncker I, Christensen L, Ostergaard JR, Jensen PK
• [In Process Citation]
• Ugeskr Laeger. 2001; 163: 4862-7
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• Fibroblast growth factors are structurally related proteins associated with cell growth, differentiation, migration, wound healing, angiogenesis, and oncogenesis. At the cellular level, their function is mediated by transmembrane tyrosinekinase receptors, fibroblast growth factor receptors. Four genes encoding fibroblast growth factor receptors have been identified, and mutations in three of these, FGFR1, FGFR2, and FGFR3, can cause different congenital, autosomal dominant disorders affecting the craniofacial and skeletal development: craniosynostosis and chondrodysplasias. The craniosynostosis syndromes: Apert syndrome, Beare-Stevenson syndrome, Crouzon syndrome, Jackson-Weiss syndrome, Muenke syndrome, Pfeiffer syndrome and Saethre-Chotzen syndrome can be caused by mutation in either FGFR1, FGFR2, or FGFR3. Saethre-Chotzen syndrome can also be caused by mutation in a functionally related gene, ACS. The same mutation can cause different syndromes, and the same syndrome can be caused by mutations in different genes. The chondrodysplasias: achondroplasia, hypochondroplasia, and thanatophoric dysplasia are all caused by mutations in FGFR3.

• Colvin JS, Green RP, Schmahl J, Capel B, Ornitz DM
• Male-to-female sex reversal in mice lacking fibroblast growth factor 9.
• Cell. 2001; 104: 875-89
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• Fgfs direct embryogenesis of several organs, including the lung, limb, and anterior pituitary. Here we report male-to-female sex reversal in mice lacking Fibroblast growth factor 9 (Fgf9), demonstrating a novel role for FGF signaling in testicular embryogenesis. Fgf9(-/-) mice also exhibit lung hypoplasia and die at birth. Reproductive system phenotypes range from testicular hypoplasia to complete sex reversal, with most Fgf9(-/-) XY reproductive systems appearing grossly female at birth. Fgf9 appears to act downstream of Sry to stimulate mesenchymal proliferation, mesonephric cell migration, and Sertoli cell differentiation in the embryonic testis. While Sry is found only in some mammals, Fgfs are highly conserved. Thus, Fgfs may function in sex determination and reproductive system development in many species.

• Lazarous DF
• The Fibroblast Growth Factors and Angiogenesis: Basic Considerations.
• Curr Interv Cardiol Rep. 2001; 3: 213-217
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• The fibroblast growth factors (FGFs) are a family of potent heparin-binding polypeptides that influence the growth, differentiation, and migration of many cell types. The FGFs are angiogenic and can induce new vessel growth. Their ability to induce new vessel growth is important in normal embryonic development, physiologic processes, pathologic processes such as malignancy, and many disease states. This article focuses on the basic aspects of FGFs in angiogenesis and discusses early clinical applications in this field.

• Hiriart M, Vidaltamayo R, Sanchez-Soto MC
• Nerve and fibroblast growth factors as modulators of pancreatic beta cell plasticity and insulin secretion.
• Isr Med Assoc J. 2001; 3: 114-6
• Display abstract

• Trophic factors such as nerve and fibroblast growth factors are important modulators of beta cell physiology. These two factors induce the extension of neurite-like processes in primary cultures of adult rat beta cells. Moreover, both NGF and FGF enhance glucose-induced insulin secretion. Since beta cells synthesize NGF and pancreatic islet cells produce FGFs, it is possible that autocrine/paracrine interactions may be major regulators of insulin secretion, and impairment of these interactions could lead to pathological states such as diabetes mellitus.

• Faux CH, Turnley AM, Epa R, Cappai R, Bartlett PF
• Interactions between fibroblast growth factors and Notch regulate neuronal differentiation.
• J Neurosci. 2001; 21: 5587-96
• Display abstract

• The differentiation of precursor cells into neurons has been shown to be influenced by both the Notch signaling pathway and growth factor stimulation. In this study, the regulation of neuronal differentiation by these mechanisms was examined in the embryonic day 10 neuroepithelial precursor (NEP) population. By downregulating Notch1 expression and by the addition of a Delta1 fusion protein (Delta Fc), it was shown that signaling via the Notch pathway inhibited neuron differentiation in the NEP cells, in vitro. The expression of two of the Notch receptor homologs, Notch1 and Notch3, and the ligand Delta1 in these NEP cells was found to be influenced by a number of different growth factors, indicating a potential interaction between growth factors and Notch signaling. Interestingly, none of the growth factors examined promoted neuron differentiation; however, the fibroblast growth factors (FGFs) 1 and 2 potently inhibited differentiation. FGF1 and FGF2 upregulated the expression of Notch and decreased expression of Delta1 in the NEP cells. In addition, the inhibitory response of the cells to the FGFs could be overcome by downregulating Notch1 expression and by disrupting Notch cleavage and signaling by the ablation of the Presenilin1 gene. These results indicate that FGF1 and FGF2 act via the Notch pathway, either directly or indirectly, to inhibit differentiation. Thus, signaling through the Notch receptor may be a common regulator of neuronal differentiation within the developing forebrain.

• Hanneken A
• Structural characterization of the circulating soluble FGF receptors reveals multiple isoforms generated by secretion and ectodomain shedding.
• FEBS Lett. 2001; 489: 176-81
• Display abstract

• Soluble fibroblast growth factor receptors (FGFRs) have been identified in multiple biological fluids, including blood. Efforts to examine the biological properties of these proteins have been hampered by the incomplete chemical characterization of the receptors within the second half of the third immunoglobulin (Ig)-like domain, where alternative splicing leads to receptor variants with different ligand binding properties. Using mass spectrometry techniques, we have mapped the soluble FGFRs to the secreted receptor, FGFR1(IIIa), the two and three Ig-like domain isoforms of FGFR1(IIIc) and a carboxyl-terminal cleavage peptide from the two and three Ig-like domain isoforms of FGFR1(IIIb). The secreted FGFR is produced by the translation of an alternatively spliced transcript and the cleaved receptors are released by ectodomain shedding of the transmembrane receptors.

• Selva EM et al.
• Dual role of the fringe connection gene in both heparan sulphate and fringe-dependent signalling events.
• Nat Cell Biol. 2001; 3: 809-15
• Display abstract

• The precise regulation of growth factor signalling is crucial to the molecular control of development in Drosophila. Post-translational modification of signalling molecules is one of the mechanisms that modulate developmental signalling specificity. We describe a new gene, fringe connection (frc), that encodes a nucleotide-sugar transporter that transfers UDP-glucuronic acid, UDP-N-acetylglucosamine and possibly UDP-xylose from the cytoplasm into the lumen of the endoplasmic reticulum/Golgi. Embryos with the frc mutation display defects in Wingless, Hedgehog and fibroblast growth factor signalling. Clonal analysis shows that fringe-dependent Notch signalling is disrupted in frc mutant tissue.

• Thomson AA
• Role of androgens and fibroblast growth factors in prostatic development.
• Reproduction. 2001; 121: 187-95
• Display abstract

• This review focuses on the role of androgens and fibroblast growth factors (FGFs) in regulating the growth and development of the prostate. It is known that androgens and mesenchymal-epithelial interactions are required for the formation and growth of the prostate, but little is known of the molecular mediators regulating prostatic organogenesis. Paracrine signalling from the mesenchyme to the epithelium is a key element of prostatic development and the action of androgens in mesenchymal cells is essential for prostatic development. This finding has led to the hypothesis that androgens regulate the expression of paracrine-acting growth factors. Although several families of growth factors play a role in regulating prostatic growth, the FGF family contains members that have been studied most comprehensively in regard to prostatic growth and branching morphogenesis. The role of FGFs in prostatic development is described in detail, since two members of the FGF family function as mesenchymal paracrine-acting factors in the prostate. It has been shown that FGF7 and FGF10 play important roles during prostatic development yet they do not appear to be regulated directly by androgens. Current models propose that growth factor expression (including FGF7 and 10) is regulated directly by androgens. However, it is possible that androgen regulation is indirect and a model outlining indirect androgen regulation of growth factors is proposed.

• Prokop A et al.
• Towards retrievable vascularized bioartificial pancreas: induction and long-lasting stability of polymeric mesh implant vascularized with the help of acidic and basic fibroblast growth factors and hydrogel coating.
• Diabetes Technol Ther. 2001; 3: 245-61
• Display abstract

• We seek to improve existing methodologies for allogenic grafting of pancreatic islets. The lack of success of encapsulated transplanted islets inside the peritoneal cavity is presently attributed to poor vascularization of the implant. A thick, fibrotic capsule often surrounds the graft, limiting survival. We have tested the hypothesis that neovascularization of the graft material can be induced by the addition of proper angiogenic factors embedded within a polymeric coat. Biocompatible and nonresorbable meshes coated with hydrophilic polymers were implanted in rats and harvested after 1-, 6-, and 12-week intervals. The implant response was assessed by histological observations on the degree of vascularity, fibrosis, and inflammation. Macrostructural geometry of meshes was conducive to tissue ingrowth into the interstitial space between the mesh filaments. Hydrogel coating with incorporated acidic or basic FGF in an electrostatic complex with polyelectrolytes and/or with heparin provided a sustained slow release of the angiogenic growth factor. Anti-factor VIII and anti-collagen type IV antibodies and a GSL I-B4 lectin were used to measure the extent of vascularization. Vigorous and persistent vascularization radiated several hundred microns from the implant. The level of vascularization should provide a sufficient diffusion of nutrients and oxygen to implanted islets. Based on our observations, stable vascularization may require a sustained angiogenic signal to allow for the development of a permanent implant structure.

• Bachler M, Neubuser A
• Expression of members of the Fgf family and their receptors during midfacial development.
• Mech Dev. 2001; 100: 313-6
• Display abstract

• Members of the FGF family play diverse roles in patterning, cell proliferation and differentiation during embryogenesis. To begin to address their function during craniofacial development we have analyzed the expression of 18 members of the Fgf family (Fgf1-15, -17, -18 and -20) and the four members of the FGF-receptor family in the prospective midfacial region between E9.5 and E11.5 by whole-mount in situ hybridization. We show that at E9.5, Fgf3, -8, -9, -10 and -17 are broadly expressed in midfacial ectoderm. Concomitant with the outgrowth of the nasal processes at E10.5, expression of Fgf3, -8, -9, -10, -15, -17 and -18 was detected in spatially restricted regions of ectoderm at the edge of the nasal pit and at the oral edge of the medial nasal process. Expression of Fgf8, Fgf9, Fgf10 and Fgf17 was still observed in these domains at E11.5. In contrast to the restricted expression patterns of the ligands, FgfR1 and FgfR2 were broadly expressed in facial mesenchyme and ectoderm, respectively, indicating a wide competence of midfacial tissue to respond to FGF signaling.

• Liu Cj, Dib-Hajj SD, Waxman SG
• Fibroblast growth factor homologous factor 1B binds to the C terminus of the tetrodotoxin-resistant sodium channel rNav1.9a (NaN).
• J Biol Chem. 2001; 276: 18925-33
• Display abstract

• In this study we demonstrate a direct interaction between a cytosolic fibroblast growth factor family member and a sodium channel. A yeast two-hybrid screen for proteins that associate with the cytoplasmic domains of the tetrodotoxin-resistant sodium channel rNa(v)1.9a (NaN) led to the identification of fibroblast growth factor homologous factor 1B (FHF1B), a member of the fibroblast growth factor family, as an interacting partner of rNa(v)1.9a. FHF1B selectively interacts with the C-terminal region but not the other four intracellular segments of rNa(v)1.9a. FHF1B binds directly to the C-terminal polypeptide of rNa(v)1.9a both in vitro and in mammalian cell lines. The N-terminal 5-77 amino acid residues of FHF1B are essential for binding to rNa(v)1.9a. FHF1B did not interact with C termini of two other sodium channels hNa(v)1.7a (hNaNE) and rNa(v)1.8a (SNS), which share 50% similarity to the C-terminal polypeptide of rNa(v)1.9a. FHF1B is the first growth factor found to bind specifically to a sodium channel. Although the functional significance of this interaction is not clear, FHF1B may affect the rNa(v)1.9a channel directly or by recruiting other proteins to the channel complex. Alternatively, it is possible that rNa(v)1.9a may help deliver this factor to the cell membrane, where it exerts its function.

• Loo BB et al.
• Production and characterization of the extracellular domain of recombinant human fibroblast growth factor receptor 4.
• Int J Biochem Cell Biol. 2000; 32: 489-97
• Display abstract

• Among the members of the fibroblast growth factor receptor family the FGFR4 has demonstrated strong dependence on heparin-like material for its activation by fibroblast growth factors. We have produced and characterized a recombinant human FGFR4 extracellular domain (FGFR4ed), in order to study its biochemical properties in isolated conditions. The FGFR4ed was expressed in an insect cell system and purified from the culture medium by Ni(2+)-affinity and gel filtration chromatography. Pure FGFR4ed was tested for FGF- and heparin-binding by covalent crosslinking experiments and by biosensor analysis. In solution, FGFR4ed formed complexes with acidic FGF (FGF-1) and basic FGF (FGF-2), both in the presence and absence of heparin. Immobilized FGFR4 also bound FGF-8 besides FGF-1 and FGF-2. Furthermore, heparin alone induced receptor oligomerization on the surface of the receptor coupled chip. Thus, the recombinant FGFR4ed revealed properties described for the cellular form of this receptor and can be used for interaction studies.

• Boilly B, Vercoutter-Edouart AS, Hondermarck H, Nurcombe V, Le Bourhis X
• FGF signals for cell proliferation and migration through different pathways.
• Cytokine Growth Factor Rev. 2000; 11: 295-302
• Display abstract

• FGFs are pleiotropic growth factors that control cell proliferation, migration and differentiation. However, FGF transduction studies have so far focused primarily on the mitogenic effect of this growth factor family and it has been difficult to assess if the described intracellular signaling pathways are dedicated solely to cell proliferation, or whether they are equally important for the migratory activity often seen in responsive cells. We review here papers in which the migratory effects of this growth factor family were clearly discriminated from proliferative effects. In toto, these studies suggest that cells use different signaling pathways for migration, such as Src and p38 MAP kinase, from those for proliferation, which tend to upregulate the ERKs. Which signaling pathway a cell uses for proliferation or migration appears to depend on many factors, including the structure and the quantity of available FGF trapped in the basal lamina by heparan sulfate co-factors, the disposition of cognate high affinity receptors and the general environment of the cell. Thus the density of the cell population, the state of the cell cycle, the presence of other factors or receptors will modulate the migratory response of cells to FGF.

• Noji S, Ohuchi H
• [Functions of fibroblast growth factor and their receptors during morphogenesis]
• Tanpakushitsu Kakusan Koso. 2000; 45: 2094-9

• Reifers F, Adams J, Mason IJ, Schulte-Merker S, Brand M
• Overlapping and distinct functions provided by fgf17, a new zebrafish member of the Fgf8/17/18 subgroup of Fgfs.
• Mech Dev. 2000; 99: 39-49
• Display abstract

• Members of the fibroblast growth factor (Fgf) family are important signaling molecules in several inductive and patterning processes, and act as brain organizer-derived signals during formation of the early vertebrate nervous system. We isolated a new member of the Fgf8/17/18 subgroup of Fgfs from the zebrafish, and studied its expression and function during somitogenesis, optic stalk and midbrain-hindbrain boundary (MHB) development. In spite of a slightly higher aminoacid similarity to Fgf8, expression analysis and mapping to a chromosome stretch that is syntenic with mammalian chromosomes shows that this gene is orthologous to mammalian Fgf17. These data provide a further example of conserved chromosomal organization between zebrafish and mammalian genomes. Using an mRNA injection assay, we show that fgf17 can act similar to fgf8 during gastrulation, when fgf17 is not normally expressed. Direct comparison of the expression patterns of fgf17 and fgf8 suggest however a possible cooperation of these Fgfs at later stages in several tissues requiring Fgf signaling. Analysis of zebrafish MHB mutants demonstrates a gene-dosage dependent requirement of fgf17 expression for the no isthmus// pax2.1 gene, showing that no isthmus/pax2.1 functions upstream of fgf17 at the MHB in a haplo-insufficient manner, similar to what has been reported for mammalian pax2 mutants. In contrast, only maintenance of fgf17 expression is disturbed at the MHB of acerebellar/fgf8 mutants. Consistent with a requirement for fgf8 function, implantation of FGF8-soaked beads induces fgf17 expression, and expression is upregulated in aussicht mutants, which display upregulation of the Fgf8 signaling pathway. Taken together, our results argue that Fgf8 and Fgf17 act as hierarchically organized signaling molecules during development of the MHB organizer and possibly other organizers in the developing nervous system.

• Ornitz DM
• FGFs, heparan sulfate and FGFRs: complex interactions essential for development.
• Bioessays. 2000; 22: 108-12
• Display abstract

• Fibroblast growth factors (FGFs) comprise a large family of developmental and physiological signaling molecules. All FGFs have a high affinity for the glycosaminoglycan heparin and for cell surface heparan sulfate proteoglycans. A large body of biochemical and cellular evidence points to a direct role for heparin/heparan sulfate in the formation of an active FGF/FGF receptor signaling complex. However, until recently there has been no direct demonstration that heparan is required for the biological activity of FGF in a developmental system in vivo. A recent paper by Lin et al.(1) has broken through this barrier to demonstrate that heparan sulfate is essential for FGF function during Drosophila development. The establishment of a role for heparan sulfate in FGFR activation in vivo suggests that tissue-specific differences in the structure of heparan may modulate the activity of FGF. BioEssays 22:108-112, 2000.

• Kato S, Sekine K
• [FGF10, a key factor for limb and lung formation]
• Seikagaku. 2000; 72: 288-91

• Sobetzko D, Braga S, Rudeberg A, Superti-Furga A
• Achondroplasia with the FGFR3 1138g-->a (G380R) mutation in two sibs sharing a 4p haplotype derived from their unaffected father.
• J Med Genet. 2000; 37: 958-9

• Mason I, Chambers D, Shamim H, Walshe J, Irving C
• Regulation and function of FGF8 in patterning of midbrain and anterior hindbrain.
• Biochem Cell Biol. 2000; 78: 577-84
• Display abstract

• In this article, an adjunct to a platform presentation at the Winternational 2000 Symposium, we summarize the recent findings of this group concerning the regulation and functions of FGF8 expressed at the isthmus of the developing brain. We show that several different FGF8 isoforms, ectopically expressed in midbrain or posterior forebrain, are able to mimic the proliferative and patterning functions previously attributed to the isthmus in tissue grafting studies. Moreover, we also show that FGF8 protein is sufficient to induce an ectopic isthmic organiser (Fgf-8+, Gbx2+) in anterior midbrain. We also provide evidence that isthmic FGF8 patterns anterior hindbrain, repressing Hox-a2 expression and setting aside a territory of the brain that includes the cerebellar anlage. We show that these effects of FGF8 are likely to be mediated via FGFR1 and be modulated by the putative FGF antagonist, Sprouty2, identified using a differential display screen. Finally, we provide evidence that the onset of Fgf8 expression is regulated by En1 and that its expression at the isthmus is subsequently maintained by a specific and direct interaction between rhombomere 1 and midbrain.

• Henderson S, Sillence D, Loughlin J, Bennetts B, Sykes B
• Germline and somatic mosaicism in achondroplasia.
• J Med Genet. 2000; 37: 956-8

• Gerwins P, Skoldenberg E, Claesson-Welsh L
• Function of fibroblast growth factors and vascular endothelial growth factors and their receptors in angiogenesis.
• Crit Rev Oncol Hematol. 2000; 34: 185-94
• Display abstract

• Angiogenesis, formation of new vessels from pre-existing ones, results from stimulation of endothelial cells, which line the vessel wall. These cells will leave their resting state and start to digest the basement membrane, proliferate, migrate and eventually differentiate to form a hollow tube. All these steps can be induced by growth factors and this review will focus on two important types of angiogenic growth factors, vascular endothelial growth factor (VEGF; also denoted vascular permeability factor, VPF) and fibroblast growth factor (FGF). Both types of factors bind to cell surface expressed receptors, which are ligand-stimulatable tyrosine kinases. Binding of the growth factors to their receptors leads to activation of the intrinsic tyrosine kinase and signal transduction to downstream signalling cascades. This results in transcriptional changes and biological responses. The molecular aspects of signalling cascades critical for endothelial cell proliferation and migration are beginning to be delineated. In contrast, signalling cascades leading to endothelial cell differentiation remain to be determined. Angiogenesis is essential for a number of physiological events such as embryonic development, ovulation, and wound healing. It has become increasingly clear that a number of diseases depend on angiogenesis. For future development of therapeutic tools, it is important to understand the molecular mechanisms that regulate angiogenesis.

• Botta M, Manetti F, Corelli F
• Fibroblast growth factors and their inhibitors.
• Curr Pharm Des. 2000; 6: 1897-924
• Display abstract

• Fibroblast growth factors (FGFs) are members of a family of polypeptides synthesized by a variety of cell types during the processes of embryonic development and in adult tissues. FGFs have been detected in normal and malignant cells and show a biological profile that includes mitogenic and angiogenic activity with a consequent crucial role in cell differentiation and development. To activate signal transduction pathways, FGFs use a dual receptor system based on tyrosine kinases and heparan sulfate (HS) proteoglycans. Based on these considerations, a variety of inhibitors able to block the interactions between FGFs and their receptors have been designed and investigated for their biological properties related to antiangiogenesis and antitumor activity. In this paper, in addition to an extensive description of the FGF family members, we report several compounds acting as FGF inhibitors by direct interaction with the growth factors. Suramin and other diverse polyanionic polysulfated and polysulfonated compounds are described, with a particular focus on suradistas. For this class of molecules, by means of molecular modeling procedures, a binding model to FGF-2 has been proposed and the structure-activity relationships of suradistas have been analyzed on the basis of the computational model described.

• Powers CJ, McLeskey SW, Wellstein A
• Fibroblast growth factors, their receptors and signaling.
• Endocr Relat Cancer. 2000; 7: 165-97
• Display abstract

• Fibroblast growth factors (FGFs) are small polypeptide growth factors, all of whom share in common certain structural characteristics, and most of whom bind heparin avidly. Many FGFs contain signal peptides for secretion and are secreted into the extracellular environment, where they can bind to the heparan-like glycosaminoglycans (HLGAGs) of the extracellular matrix (ECM). From this reservoir, FGFs may act directly on target cells, or they can be released through digestion of the ECM or the activity of a carrier protein, a secreted FGF binding protein. FGFs bind specific receptor tyrosine kinases in the context of HLGAGs and this binding induces receptor dimerization and activation, ultimately resulting in the activation of various signal transduction cascades. Some FGFs are potent angiogenic factors and most play important roles in embryonic development and wound healing. FGF signaling also appears to play a role in tumor growth and angiogenesis, and autocrine FGF signaling may be particularly important in the progression of steroid hormone-dependent cancers to a hormone-independent state.

• Antoine M et al.
• NH2-terminal cleavage of xenopus fibroblast growth factor 3 is necessary for optimal biological activity and receptor binding.
• Cell Growth Differ. 2000; 11: 593-605
• Display abstract

• Fibroblast growth factor 3 (FGF3) was originally identified as the mouse proto-oncogene Int-2, which is activated by proviral insertion in tumors induced by mouse mammary tumor virus. To facilitate the biological characterization of the ligand, we have analyzed its homologue in Xenopus laevis, XFGF3. Here we confirm that the X. laevis genome contains two distinct FGF3 alleles, neither of which is capable of encoding the NH2-terminally extended forms specified by the mouse and human FGF3 genes. Unlike the mammalian proteins, XFGF3 is efficiently secreted as a Mr 31,000 glycoprotein, gp31, which undergoes proteolytic cleavage to produce an NH2-terminally truncated product, gp27. Processing removes a segment of 18 amino acids immediately distal to the signal peptide that is not present in the mammalian homologues. By inserting an epitope-tag adjacent to the cleavage site, we show that a substantial amount of the gp27 is generated intracellularly, although processing can also occur in the extracellular matrix. Two residues are also removed from the COOH terminus. To compare the biological properties of the different forms, cDNAs were constructed that selectively give rise to the larger, gp31, or smaller, gp27, forms of XFGF3. As judged by their ability to cause morphological transformation of NIH3T3 cells, their mitogenicity on specific cell types, and their affinity for the IIIb and IIIc isoforms of Xenopus FGF receptors, gp27 has a much higher biological activity than gp31. Sequence comparison revealed an intriguing similar cleavage motif immediately downstream of the signal peptide cleavage site in the NH2-terminus of mouse and human FGF3. Analysis of secreted mutant mouse FGF3 confirmed an additional NH2-terminal processing at the corresponding sequence motif. NH2-terminal trimming of Xenopus and mammalian FGF3s may therefore be a prerequisite of optimal biological activity.

• Garcia-Alonso L, Romani S, Jimenez F
• The EGF and FGF receptors mediate neuroglian function to control growth cone decisions during sensory axon guidance in Drosophila.
• Neuron. 2000; 28: 741-52
• Display abstract

• Cell adhesion molecules (CAMs) implement the process of axon guidance by promoting specific selection and attachment to substrates. We show that, in Drosophila, loss-of-function conditions of either the Neuroglian CAM, the FGF receptor coded by the gene heartless, or the EGF receptor coded by DER display a similar phenotype of abnormal substrate selection and axon guidance by peripheral sensory neurons. Moreover, neuroglian loss-of-function phenotype can be suppressed by the expression of gain-of-function conditions of heartless or DER. The results are consistent with a scenario where the activity of these receptor tyrosine kinases is controlled by Neuroglian at choice points where sensory axons select between alternative substrates for extension.

• Wilson R, Leptin M
• Fibroblast growth factor receptor-dependent morphogenesis of the Drosophila mesoderm.
• Philos Trans R Soc Lond B Biol Sci. 2000; 355: 891-5
• Display abstract

• The Drosophila fibroblast growth factor (FGF) receptors Heartless and Breathless are required for the morphogenesis of the mesoderm and the tracheal system. In this article we discuss a number of questions relating to the morphogenesis of these tissues and speculate on poorly understood aspects of the underlying mechanisms. As yet a ligand has not been identified for Heartless, but in the case of Breathless the ligand may in some situations act as a chemotactic signal. We consider it unlikely that release of a distant chemotactic signal plays a role in the morphogenesis of the mesoderm. Instead we propose that the change in the mesoderm from an invaginated epithelium to a single layer of cells spread out on the ectoderm could be a result of the mesodermal cells trying to maximize their contact with the ectoderm. Exactly how the activation of the FGF receptors affects cell behaviour and leads to cell movement is not understood. The signal could simply be permissive, causing cells to become motile, or it could act as a directional signal for cells that are already motile, or perhaps provide both functions. Furthermore, it is unclear how signal transduction is coupled to morphological change. It seems unlikely that activation of transcription targets is essential for cell migration and it is possible that FGF signalling may have a direct effect on the cytoskeleton independent of the activation of the mitogen-activated protein kinase cascade. Analysis of the function of dof, which encodes a cytoplasmic protein required for FGF signal transduction may provide an insight into these issues.

• Dickson C, Spencer-Dene B, Dillon C, Fantl V
• Tyrosine kinase signalling in breast cancer: fibroblast growth factors and their receptors.
• Breast Cancer Res. 2000; 2: 191-6
• Display abstract

• The fibroblast growth factors [Fgfs (murine), FGFs (human)] constitute a large family of ligands that signal through a class of cell-surface tyrosine kinase receptors. Fgf signalling has been associated in vitro with cellular differentiation as well as mitogenic and motogenic responses. In vivo, Fgfs are critical for animal development, and some have potent angiogenic properties. Several Fgfs have been identified as oncogenes in murine mammary cancer, where their deregulation is associated with proviral insertions of the mouse mammary tumour virus (MMTV). Thus, in some mammary tumours of MMTV-infected mouse strains, integration of viral genomic DNA into the somatic DNA of mammary epithelial cells was found to have caused the inappropriate expression of members of this family of growth factors. Although examination of human breast cancers has shown an altered expression of FGFs or of their receptors in some tumours, their role in the causation of breast disease is unclear and remains controversial.

• Vaccarino FM, Schwartz ML, Raballo R, Rhee J, Lyn-Cook R
• Fibroblast growth factor signaling regulates growth and morphogenesis at multiple steps during brain development.
• Curr Top Dev Biol. 1999; 46: 179-200
• Display abstract

• The fibroblast growth factor (FGF) family comprises several members with distinct patterns of expression in the developing central nervous system. FGFs regulate the early specification and the subsequent growth of central nervous system regions. These different actions require the coordinated activation of distinct sets of target genes by FGFs at the appropriate stage of development. The role of FGF2 in the growth and morphogenesis of the cerebral cortex is reviewed in detail. The cellular and molecular mechanisms that underlie the action of FGF2 on cortical development are discussed.

• Wiesmann C, de Vos AM
• Putting two and two together: crystal structure of the FGF-receptor complex.
• Structure Fold Des. 1999; 7: 2515-2515
• Display abstract

• The recently determined crystal structure of an FGF-receptor complex reveals a surprising architecture and a novel mode of receptor dimerization. The structure also elucidates the role of heparan sulfate proteoglycans in receptor activation, showing significant differences from previously proposed models.

• Friesel R, Maciag T
• Fibroblast growth factor prototype release and fibroblast growth factor receptor signaling.
• Thromb Haemost. 1999; 82: 748-54

• McConnell SJ, Thon VJ, Spinella DG
• Isolation of fibroblast growth factor receptor binding sequences using evolved phage display libraries.
• Comb Chem High Throughput Screen. 1999; 2: 155-63
• Display abstract

• A fusion protein construct consisting of the short form of human fibroblast growth factor (FGFR) fused to the heavy chain of mouse IgG1 was used to screen four phage display libraries displaying 8, 13, 38 and 43 amino acids at the amino terminus of the bacteriophage M13 gene III minor coat protein. Phage with specific FGFR binding activity were isolated from the 13, 38 and 43 mer libraries. One of the highest affinity phage clones from the 13mer library was chosen to be further evolved by oligonucleotide saturation mutagenesis. We have isolated evolved sequences that have approximately 8 times the relative binding affinity of the parent sequence. The phage clones have a minimum consensus sequence of CR/SXLLXGAPFXXXXC, where X represents positions tolerant of several amino acids. A synthetic peptide based on this sequence specifically inhibits FGF from binding to its receptor in an in vitro ELISA.

• Kato S, Sekine K
• FGF-FGFR signaling in vertebrate organogenesis.
• Cell Mol Biol (Noisy-le-grand). 1999; 45: 631-8
• Display abstract

• Fibroblast growth factor and FGF receptor (FGFR) system play significant roles in many biological events including pattern formation in many tissues during vertebrate embryogenesis at early stages. The functions of each of FGFs and their receptors have recently been revealed by a variety of approaches among species. We have recently generated FGF10-deficient mice by gene targeting. This KO mice had complete truncation of the fore-and hindlimbs and with no lung. Analyses of the embryos and marker gene expression showed that FGF10 triggers sequential events, which are essential for formations of limb and lung. Focusing on FGF10 function, the FGF-FGFR system is discussed.

• Christen B, Slack JM
• Spatial response to fibroblast growth factor signalling in Xenopus embryos.
• Development. 1999; 126: 119-25
• Display abstract

• We have examined the spatial pattern of activation of the extracellular signal-regulated protein kinase (ERK) during Xenopus development, and show that it closely resembles the expression of various fibroblast growth factors (FGFs). Until the tailbud stage of development, all ERK activation domains are sensitive to the dominant negative FGF receptor, showing that activation is generated by endogenous FGF signalling. ERK is not activated by application of other growth factors like BMP4 or activin, nor is endogenous activation blocked by the respective dominant negative receptors. This shows that various domains of FGF expression, including the periblastoporal region and the midbrain-hindbrain boundary, are also sites of FGF signalling in vivo. Wounding induces a transient (<60 minutes) activation of ERK which is not significantly reduced by the dominant negative FGF receptor. An artificial FGF source, created by injection of eFGF mRNA into cleavage stage embryos, provokes ERK activation outside of its injection site over a range of several cell diameters. The range and extent of ERK activation outside the source region is unchanged by co-injection of a dominant negative form of Ras, which blocks ERK-activation within the source. This suggests that FGF protein can diffuse over several cell diameters.

• Xu X, Weinstein M, Li C, Deng C
• Fibroblast growth factor receptors (FGFRs) and their roles in limb development.
• Cell Tissue Res. 1999; 296: 33-43
• Display abstract

• Fibroblast growth factor (FGF) receptors constitute a family of four membrane-spanning tyrosine kinases (FGFR1-4) which serve as high-affinity receptors for 17 growth factors (FGF1-17). To study functions of FGF/ FGFR signals in development, mice that carry mutations in each receptor have been created by gene targeting. Analysis of these mutant mice revealed essential functions of FGF receptors in multiple biological processes, including mesoderm induction and patterning, cell growth and migration, organ formation and bone growth. In this review we discuss recent work with FGF receptors to illustrate mechanisms, through which the FGF/FGFR signals specify vertebrate limb initiation, outgrowth and patterning.

• Holowacz T, Sokol S
• FGF is required for posterior neural patterning but not for neural induction.
• Dev Biol. 1999; 205: 296-308
• Display abstract

• Fibroblast growth factor (FGF) has been implicated in a variety of developmental processes including posterior mesoderm and neural patterning. Previous work has led to contradictory roles for FGF in neural induction and anteroposterior neural patterning. Launay et al. (Development 122, 869-880, 1996) suggested a requirement for FGF in anterior neural induction. In contrast, Kroll and Amaya (Development 122, 3173-3183, 1996) and Bang et al. (Development 124, 2075-2085, 1997) proposed that FGF is not required for early neural patterning. Here we use a loss-of-function assay to examine whether FGF is required for neural patterning in three experimental situations: (i) in Xenopus early embryos, (ii) in embryonic explants consisting of presumptive dorsal mesoderm and neurectoderm (Keller explants), and (iii) in explants of dorsal ectoderm and posterior mesoderm in which FGF signaling is specifically blocked in the ectoderm. When cultured until tailbud stages, Keller explants develop neural tissue with normal anteroposterior pattern. Overexpression of the dominant-negative FGF receptor (XFD) in Keller explants inhibited the posterior neural markers En-2, Krox-20, and HoxB9, but not the panneural marker nrp-1 and the anterior neurectodermal markers XAG-1 and Xotx-2. Similar results were seen in whole embryos, but only when XFD RNA was targeted to both the dorsal and lateral regions. In contrast, addition of FGF to Keller explants resulted in a shift of the midbrain-hindbrain boundary marker En-2 to a more anterior position normally fated to become cement gland. We also determined whether FGF is required specifically by the neurectoderm for anteroposterior neural patterning. Recombinants of dorsal ectoderm and posterior mesoderm were made in which FGF was specifically blocked in the ectoderm. Spinal cord and hindbrain markers were inhibited in these recombinants, whereas anterior markers and cement gland development were enhanced. Our results demonstrate that FGF is important for posterior development in both mesoderm and neurectoderm and that neural induction and posteriorization represent separable developmental events.

• Qiu J, Mizukami H, Brown KE
• Adeno-associated virus 2 co-receptors?
• Nat Med. 1999; 5: 467-8

• Klint P, Claesson-Welsh L
• Signal transduction by fibroblast growth factor receptors.
• Front Biosci. 1999; 4: 16577-16577
• Display abstract

• The fibroblast growth factor family, with its prototype members acidic FGF (FGF-1) and basic FGF (FGF-2), binds to four related receptor tyrosine kinases, expressed on most types of cells in tissue culture. In many respects, the FGF receptors appear similar to other growth factor receptors. Thus, dimerization of receptor monomers upon ligand binding is likely to be a requisite for activation of the kinase domains, leading to receptor trans phosphorylation. FGF receptor-1 (FGFR-1), which shows the broadest expression pattern of the four FGF receptors contains at least seven tyrosine phosphorylation sites. A number of signal transduction molecules are affected by binding with different affinities to these phosphorylation sites. The potential roles of these signal transduction molecules in FGF-induced biological responses and in pathological processes are discussed.

• Zhang N, Deuel TF
• Pleiotrophin and midkine, a family of mitogenic and angiogenic heparin-binding growth and differentiation factors.
• Curr Opin Hematol. 1999; 6: 44-50
• Display abstract

• The heparin-binding polypeptide homologs pleiotrophin and midkine are the only known members of a family of secreted growth/differentiation cytokines. Pleiotrophin and midkine are both developmentally regulated and highly conserved among species. They signal a number of physiological functions involved with angiogenesis, neuorogenesis, cell migration, and mesoderm-epithelial interactions. Constitutive expression of pleiotrophin and midkine in responsive cells support their role as "tumor growth factors" and positive regulators of tumor angiogenesis. Widespread deregulation of pleiotrophin and midkine is found in many known human cancers or their derived cell lines, and the molecular targeting of pleiotrophin to block its signaling in tumor cells has limited tumor growth and metastasis in animal models. Elucidating the molecular mechanisms of pleiotrophin and midkine action in tumorgenesis and tumor angiogenesis may lead to the identification of novel targets for tumor therapy.

• Qing K, Mah C, Hansen J, Zhou S, Dwarki V, Srivastava A
• Human fibroblast growth factor receptor 1 is a co-receptor for infection by adeno-associated virus 2.
• Nat Med. 1999; 5: 71-7
• Display abstract

• Adeno-associated virus 2 (AAV)-based vectors have gained attention as a potentially useful alternative to the more commonly used retroviral and adenoviral vectors for human gene therapy. Although AAV uses the ubiquitously expressed cell surface heparan sulfate proteoglycan (HSPG) as a receptor, the transduction efficiency of AAV vectors varies greatly in different cells and tissues in vitro and in vivo. We demonstrate here that cell surface expression of HSPG alone is insufficient for AAV infection, and that AAV also requires human fibroblast growth factor receptor 1 (FGFR1) as a co-receptor for successful viral entry into the host cell. We document that cells that do not express either HSPG or FGFR1 fail to bind AAV and, consequently, are resistant to infection by AAV. These non-permissive cells are successfully transduced by AAV vectors after stable transfections with cDNAs encoding the murine HSPG and the human FGFR1. Furthermore, AAV infection of permissive cells, known to express both FGFR1 and the epidermal growth factor receptor, is abrogated by treatment of cells with basic fibroblast growth factor, but not with epidermal growth factor. The identification of FGFR1 as a co-receptor for AAV should provide new insights not only into its role in the life cycle of AAV, but also in the optimal use of AAV vectors in human gene therapy.

• Szebenyi G, Fallon JF
• Fibroblast growth factors as multifunctional signaling factors.
• Int Rev Cytol. 1999; 185: 45-106
• Display abstract

• The fibroblast growth factor (FGF) family consists of at least 15 structurally related polypeptide growth factors. Their expression is controlled at the levels of transcription, mRNA stability, and translation. The bioavailability of FGFs is further modulated by posttranslational processing and regulated protein trafficking. FGFs bind to receptor tyrosine kinases (FGFRs), heparan sulfate proteoglycans (HSPG), and a cysteine-rich FGF receptor (CFR). FGFRs are required for most biological activities of FGFs. HSPGs alter FGF-FGFR interactions and CFR participates in FGF intracellular transport. FGF signaling pathways are intricate and are intertwined with insulin-like growth factor, transforming growth factor-beta, bone morphogenetic protein, and vertebrate homologs of Drosophila wingless activated pathways. FGFs are major regulators of embryonic development: They influence the formation of the primary body axis, neural axis, limbs, and other structures. The activities of FGFs depend on their coordination of fundamental cellular functions, such as survival, replication, differentiation, adhesion, and motility, through effects on gene expression and the cytoskeleton.

• Wrighton N, Gearing D
• Fashioning FGFs from phage?
• Nat Biotechnol. 1999; 17: 1157-8

• Jung J, Zheng M, Goldfarb M, Zaret KS
• Initiation of mammalian liver development from endoderm by fibroblast growth factors.
• Science. 1999; 284: 1998-2003
• Display abstract

• The signaling molecules that elicit embryonic induction of the liver from the mammalian gut endoderm or induction of other gut-derived organs are unknown. Close proximity of cardiac mesoderm, which expresses fibroblast growth factors (FGFs) 1, 2, and 8, causes the foregut endoderm to develop into the liver. Treatment of isolated foregut endoderm from mouse embryos with FGF1 or FGF2, but not FGF8, was sufficient to replace cardiac mesoderm as an inducer of the liver gene expression program, the latter being the first step of hepatogenesis. The hepatogenic response was restricted to endoderm tissue, which selectively coexpresses FGF receptors 1 and 4. Further studies with FGFs and their specific inhibitors showed that FGF8 contributes to the morphogenetic outgrowth of the hepatic endoderm. Thus, different FGF signals appear to initiate distinct phases of liver development during mammalian organogenesis.

• Robin NH
• Molecular genetic advances in understanding craniosynostosis.
• Plast Reconstr Surg. 1999; 103: 1060-70

• Sanak M
• [Molecular genetics of congenital skeletal dysplasias related to mutations of fibroblast growth factor receptors]
• Med Wieku Rozwoj. 1999; 3: 67-82
• Display abstract

• Fibroblast growth factor receptors belong to the family of a membrane bound tyrosine kinases, which respond to stimulation by the fibroblast growth factor family of cytokines. Signalling through fibroblast growth factor receptors regulates proliferation and differentiation of connective tissue cells. Disruption of these important processes during morphogenesis results in skeletal dysplasias. Mutations of the fibroblast growth factor receptors (1, 2 and 3) cause achondroplasia, Crouzon syndrome and Apert syndrome, which are amongst the most common congenital dysmorphic syndromes. There exists a correlation between the genotype and phenotype in fibroblast growth factor receptors related syndromes. This relation can be explained on the basis of molecular pathology of the receptor protein. An increased frequency of new mutations and exclusive paternal origin of mutated alleles is explained by the role of fibroblast growth factor signalling during spermatogenesis. The availability of molecular tests for fibroblast growth factor receptors related syndromes should facilitate the diagnostic accuracy of these pleiotropic diseases which is independent of expression variability.

• Barasch J
• FGF-2: specific activity in kidney?
• Kidney Int. 1999; 56: 1156-7

• Feng HP
• Picture story. Dimers can do the deed.
• Nat Struct Biol. 1999; 6: 909-909

• Yukawa H et al.
• In vitro growth suppression of vascular smooth muscle cells using adenovirus-mediated gene transfer of a truncated form of fibroblast growth factor receptor.
• Atherosclerosis. 1998; 141: 125-32
• Display abstract

• Vascular smooth muscle cell (VSMC) proliferation associated with arterial injury causes restenosis, which remains to be resolved in cardiovascular and ischemic cerebrovascular disease, especially after balloon angioplasty. Fibroblast growth factor (FGF) is a potent mitogen and a trophic factor for a variety of cells, including VSMCs. We constructed a replication-deficient adenovirus vector, designated AxCA delta FR, coding a truncated form of fibroblast growth factor receptor-1 (FGFR-1) gene lacking the intracellular domain to interrupt receptor-mediated FGF signaling, and examined its effect on the proliferation of primary-cultured rat VSMCs. We transferred the truncated form of the FGFR-1 gene to the VSMCs and confirmed its expression and localization in infected cells by Western blotting and immunofluorescence study. The VSMCs infected with AxCA delta FR degenerated and the proliferation of these cells was suppressed markedly by the infection with this virus in vitro. Our results suggest that the receptor-mediated signal of FGFs has an important role in VSMC proliferation and gene transfer of a truncated form of FGFR using adenoviral vector may be useful for the treatment of the diseases caused by excessive proliferation of VSMCs like restenosis after percutaneous transluminal angioplasty or carotid endoarterectomy.

• Cuevas P
• Hormone-like effects of fibroblast growth factors (FGFs) in the nervous system.
• Surg Neurol. 1998; 50: 571-3

• Celli G, LaRochelle WJ, Mackem S, Sharp R, Merlino G
• Soluble dominant-negative receptor uncovers essential roles for fibroblast growth factors in multi-organ induction and patterning.
• EMBO J. 1998; 17: 1642-55
• Display abstract

• Despite a wealth of experimental data implicating fibroblast growth factor (FGF) signaling in various developmental processes, genetic inactivation of individual genes encoding specific FGFs or their receptors (FGFRs) has generally failed to demonstrate their role in vertebrate organogenesis due to early embryonic lethality or functional redundancy. Here we show that broad mid-gestational expression of a novel secreted kinase-deficient receptor, specific for a defined subset of the FGF superfamily, caused agenesis or severe dysgenesis of kidney, lung, specific cutaneous structures, exocrine and endocrine glands, and craniofacial and limb abnormalities reminiscent of human skeletal disorders associated with FGFR mutations. Analysis of diagnostic molecular markers revealed that this soluble dominant-negative mutant disrupted early inductive signaling in affected tissues, indicating that FGF signaling is required for growth and patterning in a broad array of organs and in limbs. In contrast, transgenic mice expressing a membrane-tethered kinase-deficient FGFR were viable. Our results demonstrate that secreted FGFR mutants are uniquely effective as dominant-negative agents in vivo, and suggest that related soluble receptor isoforms expressed in wild-type mouse embryos may help regulate FGF activity during normal development.

• Le Bras S, Miralles F, Basmaciogullari A, Czernichow P, Scharfmann R
• Fibroblast growth factor 2 promotes pancreatic epithelial cell proliferation via functional fibroblast growth factor receptors during embryonic life.
• Diabetes. 1998; 47: 1236-42
• Display abstract

• Several investigators have postulated that soluble growth factors are involved in the early development of the pancreas. In many tissues in which soluble factors are implicated in development, these factors act on their target cells through tyrosine kinase receptors. Because we had some preliminary evidence that fibroblast growth factor receptors (FGFRs) were expressed in the early pancreas, we investigated the effect of fibroblast growth factors (FGFs) during embryonic pancreatic development. For that purpose, we first studied the distribution and the functionality of FGFRs during pancreatic organogenesis. FGFR1 and FGFR4 were shown to be expressed at a high level during early pancreatic development before embryonic day 16, their levels of expression decreasing thereafter. The functionality of FGFR was studied next. It was demonstrated in vitro that both FGF1 and FGF2 induce the expression of NGFI-A mRNA, a useful indicator of functional growth factor-signaling pathways. Finally, the effect of FGF2 on embryonic pancreatic epithelial cell proliferation was studied. It was shown that FGF2 induces the proliferation of pancreatic epithelial cells during embryonic life. Taken together, these data strongly suggest that FGFs are implicated in pancreatic development during embryonic life.

• Pellegrini L, Commander P, Mulloy B, Martin-Martinez M, Blundell TL, Burke DF
• The role of heparin in the complex formation between fibroblast growth factor 2 and its high affinity receptor: comparative modelling and biochemical studies.
• Biochem Soc Trans. 1998; 26: 545-9

• Hicks D
• Putative functions of fibroblast growth factors in retinal development, maturation and survival.
• Semin Cell Dev Biol. 1998; 9: 263-9
• Display abstract

• The understanding of growth factor functions in the central nervous system is of paramount importance. They participate in all stages of neural development, maintenance and survival and are implicated in many pathological states. The complexity of the regulation of these signalling molecules and the intricate structure of brain tissue has made progress towards these goals difficult. The vertebrate retina has long been utilised as a model for development and function of the CNS owing to it's accessibility, organisation and composition. This is also true for growth factor research. This review will attempt to discuss some recent data, particularly with respect to the fibroblast growth factor family and highlight where more work is required for two areas of research: (1) the physiological role(s) of growth factors in retinal development and function; and (2) the potential use of growth factors in neuroprotection.retina, photoreceptor, development, survival, growth factors in neuroprotection.

• Cohen MM Jr, Neri G
• New overgrowth syndrome and FGFR3 dosage effect.
• J Med Genet. 1998; 35: 348-9

• Tartaglia M, Saulle E, Bordoni V, Battaglia PA
• Polymorphism at position 882 of the fibroblast growth factor receptor 3 (FGFR3) gene detected by SSCP analysis.
• Mol Cell Probes. 1998; 12: 335-7

• Kornmann M, Beger HG, Korc M
• Role of fibroblast growth factors and their receptors in pancreatic cancer and chronic pancreatitis.
• Pancreas. 1998; 17: 169-75
• Display abstract

• The fibroblast growth factor (FGF) family is a group of homologous heparin-binding polypeptides that has been implicated in a variety of human neoplasms and presently includes 14 members. FGF signaling is mediated by a dual-receptor system, consisting of four high-affinity tyrosine kinase receptors, termed fibroblast growth factor receptors (FGFRs), and of low-affinity heparan sulfate proteoglycan receptors that enhance ligand presentation to the FGFRs. Several FGFs, including FGF-1, -2, -3, -4, -5, -6, and -7, and several FGFR variants, among them the 2 immunoglobulin-like form and the IIIc splice variant of FGFR-1 and the keratinocyte growth factor receptor, a splice variant of FGFR-2, are expressed in human pancreatic cancer cell lines and are overexpressed in human pancreatic cancers or in the pancreas of chronic pancreatitis and, therefore, may play important roles in the pathobiology of these pancreatic diseases. This review summarizes the current information on the involvement of the FGF family and their receptors in human pancreatic cancer and chronic pancreatitis.

• Matsui Y, Yasui N, Kimura T, Tsumaki N, Kawabata H, Ochi T
• Genotype phenotype correlation in achondroplasia and hypochondroplasia.
• J Bone Joint Surg Br. 1998; 80: 1052-6
• Display abstract

• Recent studies of the fibroblast growth factor receptor 3 (FGFR3) gene have established that achondroplasia and hypochondroplasia are allelic disorders of different mutations. To determine whether the genotype could be distinguished on the basis of the phenotype, we analysed height, arm span, and skeletal radiographs from 23 patients with achondroplasia and the G380R mutation of FGFR3 and eight with hypochondroplasia and the N540K mutation. Both conditions share the classical pathological features of micromelic short stature, reduced or unchanged interpedicular distances in the lumbar spine, disproportionately long fibulae, and squared and shortened pelvic ilia. These were significantly more severe in the G380R patients than in the N540K patients. Our findings have shown a firm statistical correlation between the genotype and the phenotype, although there were a few exceptional cases in which there was phenotypic overlap between the two conditions.

• Robin NH, Scott JA, Cohen AR, Goldstein JA
• Nonpenetrance in FGFR3-associated coronal synostosis syndrome.
• Am J Med Genet. 1998; 80: 296-7

• Burke D, Wilkes D, Blundell TL, Malcolm S
• Fibroblast growth factor receptors: lessons from the genes.
• Trends Biochem Sci. 1998; 23: 59-62
• Display abstract

• The fibroblast growth factor receptors (FGFRs) are a family of transmembrane tyrosine kinases involved in signalling via interactions with the family of fibroblast growth factors (FGFs). Genetic findings have provided a way of dissecting these interactions. Mutations in three members of the FGFR family have been found in patients with birth defects involving craniosynostosis (premature fusion of the cranial sutures) or skeletal abnormalities. Analyses of the spectrum of mutations found predict that many of them will result in ligand-independent activation of the receptors. Amino acids have also been identified that are likely to be important in determining the specificity of FGFR-FGF interactions.

• Foehr ED, Raffioni S, Fuji R, Bradshaw RA
• FGF signal transduction in PC12 cells: comparison of the responses induced by endogenous and chimeric receptors.
• Immunol Cell Biol. 1998; 76: 406-13
• Display abstract

• Rat phaeochromocytoma (PC12) cells respond to many growth factors and produce different phenotypes, including neurite outgrowth. Receptor tyrosine kinases (RTK), which activate multiple signalling pathways in response to ligand binding, initiate many of these. One such family of receptors, the fibroblast growth factor receptor (FGFR), has four different members and expresses at least three of these in PC12 cells. A chimeric tyrosine kinase receptor, consisting of the extracellular domain of human plasma-derived growth factor receptor-beta (hPDGFR-beta) and the transmembrane and intracellular region of FGFR1 (designated PFR1), was constructed and was stably transfected into cloned PC12 cell lines. This chimera, which can be activated without stimulating endogenous RTK including other FGFR, induces neurite outgrowth in a PDGF-dependent manner. By altering the protocol for preparing the retroviral vectors, cells with a wide range of expression levels can be obtained. The amount of these chimeric receptors seems to correlate with the time and the intensity of response as observed in neurite outgrowth assays. Analysis of proteins implicated in FGFR1 signalling indicates that upon stimulation, a tyrosine phosphorylated protein designated FRS2 associates with SOS, Grb2 and the receptor. The chimeric receptor appears entirely similar to that observed for the stimulation of native PC12 cells with FGF2. These results support the view that FRS2 is the dominant FGFR1 signalling entity in PC12 cells.

• Pickles JO, Harter C, Rebillard G
• Fibroblast growth factor receptor expression in outer hair cells of rat cochlea.
• Neuroreport. 1998; 9: 4093-5
• Display abstract

• Fibroblast growth factors (FGFs) are critical for normal development of the organ of Corti, and may also protect hair cells from ototoxic damage. Four different fibroblast growth factors are known, three of which have different splice variants in the extracellular immunoglobin-like (Ig) III FGF-binding domain, giving different patterns of sensitivity to the different FGFs. Analysis of a cDNA library of rat outer hair cells by the polymerase chain reaction, using isoform specific primers, showed expression only of FGF receptor 3, splice variant IIIc. This allows us to predict the pattern of sensitivity to applied FGFs, which may be useful in targeting outer hair cells selectively during an FGF-based strategy for cochlear therapy.

• Meiri KF, Saffell JL, Walsh FS, Doherty P
• Neurite outgrowth stimulated by neural cell adhesion molecules requires growth-associated protein-43 (GAP-43) function and is associated with GAP-43 phosphorylation in growth cones.
• J Neurosci. 1998; 18: 10429-37
• Display abstract

• The mechanisms whereby cell adhesion molecules (CAMs) promote axonal growth and synaptic plasticity are poorly understood. Here we show that the neurite outgrowth stimulated by NCAM-mediated fibroblast growth factor (FGF) receptor activation in cerebellar granule cells is associated with increased GAP-43 phosphorylation on serine-41. In contrast, neither NCAM nor FGF was able to stimulate neurite outgrowth in similar neurons from mice in which the GAP-43 gene had been deleted by homologous recombination. Integrin-mediated neurite outgrowth was unaffected by GAP-43 deletion. Both neurite outgrowth and rapid phosphorylation of GAP-43 in isolated growth cones required the first three Ig domains of a NCAM-Fc chimera and were stimulated maximally at 5 micrograms/ml (approximately 50 nM). Likewise, GAP-43 phosphorylation in isolated growth cones also was stimulated by an L1-Fc chimera. Both neurite outgrowth and NCAM-stimulated GAP-43 phosphorylation were inhibited by antibodies to the FGF receptor and a diacylglycerol lipase inhibitor (RHC80267) that blocks the production of arachidonic acid in response to activation of the FGF receptor. Direct activation of the FGF receptor and the arachidonic acid cascade with either basic FGF or melittin also resulted in increased GAP-43 phosphorylation. These data suggest that the stimulation of neurite outgrowth by NCAM requires GAP-43 function and that GAP-43 phosphorylation in isolated growth cones occurs via an FGF receptor-dependent increase in arachidonic acid.

• Hall JG
• A bone is not a bone is not a bone.
• J Pediatr. 1998; 133: 5-6

• Faham S, Linhardt RJ, Rees DC
• Diversity does make a difference: fibroblast growth factor-heparin interactions.
• Curr Opin Struct Biol. 1998; 8: 578-86
• Display abstract

• Fibroblast growth factors (FGFs) are members of a protein family with a broad range of biological activities. The best characterized FGFs interact with two distinct extracellular receptors--a transmembrane tyrosine kinase FGF receptor (FGFR) and a heparan f1p4ate-related proteoglycan of the extracellular matrix. These components form a FGF-FGFR-proteoglycan complex that activates the FGF-mediated signal transduction process through FGFR dimerization. Recent crystal structure determinations of FGF-heparin complexes have provided insights into both the interactions between these components and the role of heparin-like proteoglycans in FGF function. Future advances in this field will benefit enormously from an ability to specifically prepare homogeneous heparin-based oligosaccharides of defined sequence for use in biochemical and structural studies of FGF and many other systems.

• Takahashi H
• [Studies on the expression of fibroblast growth factors and fibroblast growth factor receptors in human prostate cancer]
• Nippon Hinyokika Gakkai Zasshi. 1998; 89: 836-45
• Display abstract

• PURPOSE: We tried to clarify the role of fibroblast growth factors (FGFs) and those receptors (FGF-Rs) in cell proliferation of human prostate cancer. METHODS: The mRNA expression of FGF1, FGF2, FGF7, FGF-R1, FGF-R2 (IIIb), and FGF-R2 (IIIc) was investigated by RT-PCR in androgen sensitive cells (LNCaP), androgen-independent cells (PC3) and primary cultured stromal (PS) and epithelial cells (PE) from benign prostatic hyperplasia (BPH). Expression of the mRNA of FGF-R1, FGF-R2 (IIIb) and FGF-R2 (IIIc) in human prostate cancer tissue was similarly analyzed. Furthermore, the level of FGF-R1 expression in human prostate cancer was measured by semi-quantitative RT-PCR. RESULTS: FGF-R1 mRNA was detected in LNCaP, PC3 and the primary cultured stromal cells of BPH. FGF-R2 (IIIb) was seen in LNCaP cells and the primary cultured epithelial cells of BPH, while FGF-R2 (IIIc) was only observed in PC3. FGF1 mRNA was expressed in LNCaP and PC3, while FGF2 mRNA was in PC3 alone. The expression of FGF7 mRNA was detected only in the primary cultured stromal cells. Of 17 patients with human prostate cancer, FGF-R2 (IIIb) was detected in 2 and FGF-R2 (IIIc) in 15. Histological type of two cases having FGF-R2 (IIIb) were well differentiated adenocarcinoma. The mRNA levels of FGF-R1 in poorly and moderately differentiated types were significantly higher than those in well differentiated ones (p < 0.05). CONCLUSION: These findings suggest that several changes of expression in FGFs and FGF-Rs may correlate with malignant progression of human prostate cancer.

• Herrlich P et al.
• How tumor cells make use of CD44.
• Cell Adhes Commun. 1998; 6: 141-7
• Display abstract

• A variant of CD44 containing exon v3 sequences is expressed in the apical ectodermal ridge of the limb bud during embryogenesis. This variant is modified by heparan sulfate moieties and acts as low affinity receptor for FGFs. These FGFs are presented by CD44 to mesenchymal cells which induces their proliferation and limb outgrowth. We suggest that a similar growthfactor presentation mechanism accounts for the function of CD44 variants on metastasizing tumor cells.

• Kosher RA
• Syndecan-3 in limb skeletal development.
• Microsc Res Tech. 1998; 43: 123-30
• Display abstract

• Syndecan-3 is a member of a family of heparan sulfate proteoglycans that function as extracellular matrix receptors and as co-receptors for growth factors and signalling molecules. A variety of studies indicate that syndecan-3 is involved in several aspects of limb morphogenesis and skeletal development. Syndecan-3 participates in limb outgrowth and proliferation in response to the apical ectodermal ridge; mediates cell-matrix and/or cell-cell interactions involved in regulating the onset of chondrogenesis; may be involved in regulating the onset of osteogenesis and joint formation and, plays a role in regulating the proliferation of epiphyseal chondrocytes during endochondral ossification.

• Uruno T, Komi A, Imamura T
• [Progress of signal transduction research for fibroblast growth factors]
• Seikagaku. 1998; 70: 446-53

• Mattern J, Koomagi R, Volm M
• Xenotransplantability of human cancers in mice.
• J Natl Cancer Inst. 1998; 90: 1747-9

• Naski MC, Ornitz DM
• FGF signaling in skeletal development
• Front Biosci. 1998; 3: 78194-78194
• Display abstract

• The fibroblast growth factor receptor family consists of four receptor tyrosine kinases which bind with varying affinity and specificity to a family of at least fifteen polypeptide ligands. The receptors and ligands perform many essential functions during growth, development and repair. Recent discoveries show that a growing number of skeletal abnormalities result from mutations in the fibroblast growth factor receptors. These findings have led to a greater understanding of the role of fibroblast growth factor signaling during skeletogenesis and have focused research interests on the effects of fibroblast growth factors on endochondral and intramembranous bone development.

• McKeehan WL, Wu X, Jang JH, Kan M
• High salt inhibits both heparin-dependent and heparin-independent complexes of fibroblast growth factor and the receptor kinase.
• In Vitro Cell Dev Biol Anim. 1997; 33: 727-30

• Klein S, Roghani M, Rifkin DB
• Fibroblast growth factors as angiogenesis factors: new insights into their mechanism of action.
• EXS. 1997; 79: 159-92

• Bansal R, Pfeiffer SE
• Regulation of oligodendrocyte differentiation by fibroblast growth factors.
• Adv Exp Med Biol. 1997; 429: 69-77

• Skaer H
• Morphogenesis: FGF branches out.
• Curr Biol. 1997; 7: 23841-23841
• Display abstract

• Signalling via tissue-specific receptors for fibroblast growth factor is important in tracheal branching and mesodermal patterning in Drosophila; both processes may involve a common primary response - detachment and directed migration of the target cells - that is conserved from worms to mammals.

• Jackson D, Bresnick J, Dickson C
• A role for fibroblast growth factor signaling in the lobuloalveolar development of the mammary gland.
• J Mammary Gland Biol Neoplasia. 1997; 2: 385-92
• Display abstract

• The inappropriate expression of growth factors, or activating mutations of their receptors, have been implicated as causative factors in mouse and human mammary cancer. For example, it has been known for some time that three members of the fibroblast growth factor (FGF) family behave like oncogenes in virally induced mammary cancer of mice. In normal circumstances, signaling via FGF receptors is known to mediate growth, differentiation, and patterning, during embryogenesis and fetal development. A powerful approach to dissecting the roles for these signaling pathways is to determine the developmental consequences of abrogating their function in transgenic mice. In this review, we describe the use of dominant negative FGF receptors to evaluate the contribution of specific FGF signals in normal mammary gland development. These studies have revealed that normal lobuloalveolar development requires FGF signaling to the mammary epithelium, a function that is presumably usurped by MMTV in mouse mammary tumorigenesis.

• Wilke TA, Gubbels S, Schwartz J, Richman JM
• Expression of fibroblast growth factor receptors (FGFR1, FGFR2, FGFR3) in the developing head and face.
• Dev Dyn. 1997; 210: 41-52
• Display abstract

• Fibroblast growth factors may play an important role in the differential growth of the skull, brain, and facial prominences. In order to understand the role of FGFs in vivo, we have analyzed the competency of head mesenchyme to respond to FGFs via expression of the high affinity receptors FGFR1, 2, and 3. Receptor transcripts, especially those of FGFR2 and FGFR3, were localized to specific regions of the head. We raise the possibilities of particular receptor-ligand combinations and the possible functions of these interactions in the morphogenesis of the head, face, and brain. Finally, we discuss the relationship between FGF receptor expression in the chicken and the phenotypes of FGF receptor mutations in humans.

• Ratajczak MZ
• Fibroblast growth factors and early hemopoietic cell development.
• Leuk Lymphoma. 1997; 27: 221-9
• Display abstract

• The role of fibroblast growth factors (FGFs) in the regulation of the human hematopoiesis is controversial. Older publications suggest that FGFs play an important role in regulating the proliferation of human hematopoietic progenitor and stem cells. Such studies should be interpreted with caution because they were typically carried out in the presence of serum which not only contains significant amounts of FGFs, but other cytokines as well. The development of greatly improved techniques for isolating purified populations of primitive hematopoietic cells, for culturing such cells in serum free conditions, and for analyzing the molecular consequences of exposing these cells to FGF cytokines allow the conclusions of these earlier studies to be tested more rigorously. Our investigations, and those of others, suggest that FGFs may not play an important direct role in regulating the development of human hematopoietic stem cells. These data, and our view of their implications, are discussed in this review.

• Lewandoski M, Meyers EN, Martin GR
• Analysis of Fgf8 gene function in vertebrate development.
• Cold Spring Harb Symp Quant Biol. 1997; 62: 159-68

• Gorlin RJ
• Atypical achondroplasia.
• Am J Med Genet. 1997; 70: 2022035-2022035

• Rossant J, Ciruna B, Partanen J
• FGF signaling in mouse gastrulation and anteroposterior patterning.
• Cold Spring Harb Symp Quant Biol. 1997; 62: 127-33

• Bikfalvi A, Klein S, Pintucci G, Rifkin DB
• Biological roles of fibroblast growth factor-2.
• Endocr Rev. 1997; 18: 26-45

• Lin HY, Kaplow J, Jaye M, Hayman MJ
• Ligand-binding specificity of human fibroblast growth factor receptor-3 IIIc.
• FEBS Lett. 1997; 411: 389-92
• Display abstract

• Earlier studies indicated that human fibroblast growth factor receptor (FGFR)-3 IIIc was activated equally well by both FGF-1 and FGF-2. In contrast, murine FGFR-3 IIIc was preferentially activated by FGF-1. To address this issue, we determined the ligand-binding specificity of human FGFR-3 IIIc in comparison with human FGFR-1 IIIc. By equilibrium binding human FGFR-3 IIIc preferentially bound FGF-1 with high affinity, whereas FGFR-1 IIIc bound both FGF-1 and -2 with high affinity. By competition binding using FGF-1, -2, -4, or -6, FGF-1 competed more efficiently than the other FGFs. These results suggest that like the murine FGFR-3 III, FGF-1 is a preferred ligand for human FGFR-3 IIIc.

• Galzie Z, Kinsella AR, Smith JA
• Fibroblast growth factors and their receptors.
• Biochem Cell Biol. 1997; 75: 669-85
• Display abstract

• Fibroblast growth factors (FGFs) represent a group of polypeptide mitogens eliciting a wide variety of responses depending upon the target cell type. The knowledge of the cell surface receptors mediating the effects of FGFs has recently expanded remarkably. The complexity of the FGF family and the FGF-induced responses is reflected in the diversity and redundancy of the FGF receptors. In this review, a number of biochemical characteristics and biological properties of the FGF family and its receptors are described and their expression both in normal tissues and in tumours is discussed. Finally we speculate on the targetting of growth inhibition agents to tumours through FGF receptors.

• Gorlin RJ
• Fibroblast growth factors, their receptors and receptor disorders.
• J Craniomaxillofac Surg. 1997; 25: 69-79

• Cuevas P
• Therapeutic prospects for fibroblast growth factor treatment of brain ischemia.
• Neurol Res. 1997; 19: 355-6
• Display abstract

• Ischemic stroke causes neurological disability in millions of people worldwide each year. At present, standard therapies are often ineffective in stroke prevention. In this review, we summarize recent findings on the role of fibroblast growth factors in neuroprotection after experimental brain ischemia.

• Cohen MM Jr
• A matter of reading English.
• Am J Med Genet. 1996; 63: 503-4

• Smith JC
• Transgenic frogs and FGF signalling in early development.
• Trends Genet. 1996; 12: 439-40

• Okamoto T, Tanaka Y, Kan M, Sakamoto A, Takada K, Sato JD
• Expression of fibroblast growth factor binding protein HBp17 in normal and tumor cells.
• In Vitro Cell Dev Biol Anim. 1996; 32: 69-71

• Ornitz DM et al.
• Receptor specificity of the fibroblast growth factor family.
• J Biol Chem. 1996; 271: 15292-7
• Display abstract

• Fibroblast growth factors (FGFs) are essential molecules for mammalian development. The nine known FGF ligands and the four signaling FGF receptors (and their alternatively spliced variants) are expressed in specific spatial and temporal patterns. The activity of this signaling pathway is regulated by ligand binding specificity, heparan sulfate proteoglycans, and the differential signaling capacity of individual FGF receptors. To determine potentially relevant ligand-receptor pairs we have engineered mitogenically responsive cell lines expressing the major splice variants of all the known FGF receptors. We have assayed the mitogenic activity of the nine known FGF ligands on these cell lines. These studies demonstrate that FGF 1 is the only FGF that can activate all FGF receptor splice variants. Using FGF 1 as an internal standard we have determined the relative activity of all the other members of the FGF family. These data should serve as a biochemical foundation for determining developmental, physiological, and pathophysiological processes that involve FGF signaling pathways.

• Bellus GA, McIntosh I, Szabo J, Aylsworth A, Kaitila I, Francomano CA
• Hypochondroplasia: molecular analysis of the fibroblast growth factor receptor 3 gene.
• Ann N Y Acad Sci. 1996; 785: 182-7

• Alderborn A, Anvret M, Gustavson KH, Hagenas L, Wadelius C
• Achondroplasia in Sweden caused by the G1138A mutation in FGFR3.
• Acta Paediatr. 1996; 85: 1506-7
• Display abstract

• Achondroplasia, an autosomal dominant inherited disorder, is one of the most common forms of skeletal dysplasia resulting in disproportionate extreme shortness. Recently, two point mutations, both affecting nucleotide 1138 in the fibroblast growth factor receptor type 3 (FGFR3) gene, were found to be the cause of the disorder. We investigated DNA from 16 Swedish patients with achondroplasia for the presence of these mutations. All patients were found to be heterozygous for the G to A transition at nucleotide 1138. Our data thus support previous reports showing a striking genetic homogeneity, in that almost all achondroplasia patients have the FGFR3 G380R mutation at the protein level.

• Skovby F, Hertz H
• [Fibroblast growth factors, their receptors and congenital diseases]
• Ugeskr Laeger. 1996; 158: 2865-2865

• Green PJ, Walsh FS, Doherty P
• Promiscuity of fibroblast growth factor receptors.
• Bioessays. 1996; 18: 639-46
• Display abstract

• Fibroblast growth factor receptors (FGFRs) have been implicated in many developmental and regenerative events, including axial organisation, mesodermal patterning, keratinocyte organisation and brain development. The consensus view that this reflects a role for one or other of the nine known members of the fibroblast growth factor family in these processes has recently been challenged by the suggestion that FGFRs might be directly activated by a much wider range of ligands, including heparan sulphate proteoglycans and neural cell adhesion molecules. In addition, two novel soluble ligands for FGFRs have been identified using yeast two-hybrid technology. Overall, the new findings suggest that in terms of ligand binding the FGFRs might be an even more promiscuous family of receptor tyrosine kinases than was already appreciated.

• Mason I
• Neural induction: do fibroblast growth factors strike a cord?
• Curr Biol. 1996; 6: 672-5
• Display abstract

• Recent studies suggest that fibroblast growth factors (FGFs), or FGF receptor-mediated signalling, function in specifying posterior identity in the developing neural tube, and possibly also in neural induction.

• Smallwood PM et al.
• Fibroblast growth factor (FGF) homologous factors: new members of the FGF family implicated in nervous system development.
• Proc Natl Acad Sci U S A. 1996; 93: 9850-7
• Display abstract

• Four new members of the fibroblast growth factor (FGF) family, referred to as fibroblast growth factor homologous factors (FHFs), have been identified by a combination of random cDNA sequencing, data base searches, and degenerate PCR. Pairwise comparisons between the four FHFs show between 58% and 71% amino acid sequence identity, but each FHF shows less than 30% identity when compared with other FGFs. Like FGF-1 (acidic FGF) and FGF-2 (basic FGF), the FHFs lack a classical signal sequence and contain clusters of basic residues that can act as nuclear localization signals. In transiently transfected 293 cells FHF-1 accumulates in the nucleus and is not secreted. Each FHF is expressed in the developing and adult nervous systems, suggesting a role for this branch of the FGF family in nervous system development and function.

• Hicks D
• Characterization and possible roles of fibroblast growth factors in retinal photoreceptor cells.
• Keio J Med. 1996; 45: 140-54
• Display abstract

• There is increasing evidence that soluble polypeptide growth factors such as those belonging to the fibroblast growth factor (FGF) family play important roles in many aspects of photoreceptor cell biology, including differentiation, continued survival and pathology. At least two members of this family, acidic FGF and basic FGF, are synthesized by, bind to and have profound effects upon these highly specialized retinal first-order neurons. The present review presents an overview of the evidence accumulated to date and will try to suggest future directions for research.

• Goldfarb M
• Functions of fibroblast growth factors in vertebrate development.
• Cytokine Growth Factor Rev. 1996; 7: 311-25
• Display abstract

• Fibroblast growth factors (FGFs) are a class of secreted polypeptide ligands which mediate diverse cellular responses during embryonic, fetal, and postnatal vertebrate development. The purposes of this review are to provide a condensed overview of FGFs and their receptors, to catalog and categorize the functions of FGFs in vertebrate development, to present recent discoveries relating to the interplay of FGFs with other secreted ligands in the control of tissue growth and patterning, and to discuss several potential directions for future research in the field.

• Malcolm S, Reardon W
• Fibroblast growth factor receptor-2 mutations in craniosynostosis.
• Ann N Y Acad Sci. 1996; 785: 164-70

• Tanaka EM, Gann AF
• Limb development. The budding role of FGF.
• Curr Biol. 1995; 5: 594-7
• Display abstract

• Implantation of beads soaked in fibroblast growth factor into the flank of a chick embryo causes extra limbs to be formed, suggesting that FGF is important in initiating limb buds.

• Hecht JT, Herrera CA, Greenhaw GA, Francomano CA, Bellus GA, Blanton SH
• Confirmatory linkage of hypochondroplasia to chromosome arm 4p.
• Am J Med Genet. 1995; 57: 505-6

• Lajeunie E, Ma HW, Bonaventure J, Munnich A, Le Merrer M, Renier D
• FGFR2 mutations in Pfeiffer syndrome.
• Nat Genet. 1995; 9: 108-108

• Gillespie LL, Chen G, Paterno GD
• Cloning of a fibroblast growth factor receptor 1 splice variant from Xenopus embryos that lacks a protein kinase C site important for the regulation of receptor activity.
• J Biol Chem. 1995; 270: 22758-63
• Display abstract

• A cDNA clone, predicted to encode a variant form of the type 1 fibroblast growth factor receptor (FGFR1) containing a dipeptide Val-Thr (VT) deletion at amino acid positions 423 and 424 located within the juxtamembrane region, was isolated from a Xenopus embryo (stage 8 blastula) library. Sequence analysis of genomic DNA encoding a portion of the FGFR1 juxtamembrane region demonstrated that this variant form arises from use of an alternative 5' splice donor site. RNase protection analysis revealed that both VT- and VT+ forms of the FGFR1 were expressed throughout embryonic development, the VT+ being the major form. Amino acid position 424 is located within a consensus sequence for phosphorylation by a number of Ser/Thr kinases. We demonstrate that a VT+ peptide was specifically phosphorylated by protein kinase C (PKC) in vitro, but not by protein kinase A (PKA). A VT- peptide, on the other hand, was not a substrate for either enzyme. Phosphorylation levels of in vitro synthesized FGFR-VT+ protein by PKC were twice that of FGFR-VT- protein. In a functional assay, Xenopus oocytes expressing FGFR-VT- or FGFR-VT+ protein were equally able to mobilize intracellular Ca2+ in response to basic fibroblast growth factor (bFGF). However, pretreatment with phorbol 12-myristate 13-acetate significantly reduced this mobilization in oocytes expressing FGFR-VT+ while having little effect on oocytes expressing FGFR-VT-. These findings demonstrate that alternative splicing of Val423-Thr424 generates isoforms which differ in their ability to be regulated by phosphorylation and thus represents an important mechanism for regulating FGFR activity.

• Slavin J
• Fibroblast growth factors: at the heart of angiogenesis.
• Cell Biol Int. 1995; 19: 431-44
• Display abstract

• Acidic fibroblast growth factor (FGF-1) and basic fibroblast growth factor (FGF-2) are ubiquitous cytokines found in many tissues. They have effects on multiple cell types derived from mesoderm and neuroectoderm, including endothelial cells. In this review the structure and function of the fibroblast growth factor family and its receptors are described. The evidence implicating both FGF-1 and FGF-2 in the control of blood vessel formation is presented and their involvement in normal and pathological angiogenesis during adult life is then described in more detail.

• Lotto RB, Price DJ
• The stimulation of thalamic neurite outgrowth by cortex-derived growth factors in vitro: the influence of cortical age and activity.
• Eur J Neurosci. 1995; 7: 318-28
• Display abstract

• Recent in vitro experiments have provided useful insights into the development of connections between the thalamus and the cortex. While most of these previous studies focused on neurite guidance and target recognition, our experiments used a serum-free culture system to examine the possible roles of unidentified diffusible cortex-derived growth factors. We demonstrated that occipital cortical explants release diffusible growth factors that enhance neurite outgrowth from explants of the posterior thalamus (the region around the developing lateral geniculate nucleus). The amount of thalamic outgrowth was dependent on the age of the cocultured cortical slices. Our results suggest that there is an overall increase in the release of cortex-derived growth factors during the first three postnatal weeks in mice; this parallels known postnatal increases in the production of several identified growth factors. We found evidence for two peaks in the release of cortex-derived growth factors during the general upward trend, the first at around postnatal day 6 (shortly after thalamocortical innervation of layer 4) and a second between postnatal days 14 and 18 (just after eye-opening). The increased release of cortex-derived growth factors was not found when cortical slices were from mice that had been dark-reared from birth, suggesting that neural activity may be important for enhancing release. Other regions of the central nervous system, including the cerebellum and medulla, were also capable of stimulating some thalamic outgrowth; neither additional explants of the thalamus nor hepatic explants enhanced outgrowth. Fibroblast growth factor is one substance that is distributed preferentially among those tissues that were stimulatory in our experiments. Its level of transcription is known to increase in the brain during the first three postnatal weeks and to be influenced by neural activity. At low doses, fibroblast growth factor greatly increased outgrowth from isolated posterior thalamic explants. Nerve growth factor, another candidate molecule, was less effective. Overall, our results complement the in vivo observations of others on the synthesis of identified growth factors in the cortex and the factors that influence their production. They suggest that growth factors may influence thalamic neurons, and indicate that fibroblast growth factor, and possibly nerve growth factor, are two candidates for molecules mediating the in vitro effects.

• Ali J, Mansukhani A, Basilico C
• Fibroblast growth factor receptors 1 and 2 are differentially regulated in murine embryonal carcinoma cells and in response to fibroblast growth factor-4.
• J Cell Physiol. 1995; 165: 438-48
• Display abstract

• We have studied the expression of two of the receptors for fibroblast growth factors, FGFR-1 and FGFR-2, in response to ligand binding and in embryonal carcinoma (EC cells). Exposure of mouse fibroblasts to FGF-4 or FGF-2 results in a drastic downregulation of the mRNA levels for FGFR-2, while expression of FGFR-1 mRNA appears unaffected. Furthermore, FGF-4 transformed cells display low levels of FGFR-2 mRNA and these levels are significantly increased by treatment with anti FGF-4 neutralizing antibodies. In undifferentiated F9 EC cells, the levels of FGFR-2 mRNA are very low and increase substantially upon induction of differentiation. The levels of mRNA for FGFR-1 are again unaffected. To gain information on the regulation of expression of the gene encoding FGFR-2 (bek) we have cloned the FGFR-2 promoter region and used it to drive the expression of plasmids encoding the bacterial CAT enzyme. Transfection of these plasmids into FGF treated and untreated cells did not produce significant variation in CAT activity, suggesting that FGFR-2 downregulation in response to ligand binding occurs mainly by a post-transcriptional mechanism. In contrast, plasmids containing as little as 140 nt of the FGFR-2 promoter region were regulated in F9 cells, showing substantially higher expression in differentiated than in undifferentiated cells. It appears therefore that FGFR-2 expression in fibroblasts and EC cells is regulated by somewhat different mechanisms. In contrast, FGFR-1 expression does not vary substantially under the conditions shown to affect FGFR-2 expression. The implications of these findings are discussed.

• Liu HM
• Correlation between proto-oncogene, fibroblast growth factor and adaptive response in brain infarct.
• Prog Brain Res. 1995; 105: 239-44
• Display abstract

• Ischemia/hypoxia rapidly induce ischemic changes in vulnerable neurons: cortical neurons in layers II-III and V, hippocampal neurons, cerebellar Purkinje cells and certain basal ganglia and brainstem neurons. The ischemic changes are manifested histologically by nuclear pyknosis, cytoplasmic shrinkage and basophilia. These neurons exhibit strong and persistent expression of immediate early genes (IEGs): c-fos and c-jun. The onset of IEG expression is followed within a day by enhanced bFGF expression in non-ischemic neurons in the same general regions. The appearance of bFGF is followed within another day by proliferation of blood vessels, macrophages and glial cells around the infarct. The newly-formed blood vessels and macrophages migrate into the necrotic infarct aiming at disposal of the necrotic debris. The gliosis although concentrated around the infarct spreads to involve remote regions of both hemispheres. Based on the spatiotemporal correlation between cell proliferation and bFGF and the known mitogenic properties of bFGF, we believe that this molecule may be responsible for the late response in brain infarct including angiogenesis, gliosis and macrophage proliferation. The physiological roles of IEGs in the chain of adaptive response following brain infarction and its relationship with bFGF are subjects pending future investigations.

• Muenke M, Schell U
• Fibroblast-growth-factor receptor mutations in human skeletal disorders.
• Trends Genet. 1995; 11: 308-13
• Display abstract

• Fibroblast-growth-factor receptors (FGFRs), members of the tyrosine-kinase receptor family, play a crucial role in signal transduction and development. Recently, unique mutations in three human FGFR-encoding genes (FGFR1-3) have been identified as the cause of a variety of skeletal disorders. Comparison of these specific mutations with the resulting phenotypes is now providing new insight into the role of these receptors in normal and abnormal bone development.

• Yayon A, Gray TE, Zimmer Y, Eisenstein M, Givol D
• Ligand binding by fibroblast growth factor receptors investigated using chimeric receptor molecules.
• J Recept Signal Transduct Res. 1995; 15: 185-97
• Display abstract

• In order to map in detail the ligand binding sites of fibroblast growth factor receptor 2 (FGFR2) and keratinocyte growth factor receptor (KGFR), we have generated receptor molecules that are chimeric within the membrane proximal sequence that varies between them. The chimeric molecules are found to bind aFGF with a greater than 5-fold difference in affinity, indicating that there is coupling between the chimeric regions with respect to aFGF binding. Further, binding of bFGF and KGF is abolished in the chimeras, showing that the binding site for these ligands requires the whole of the 48- or 50- amino acid variable sequence to be intact. Direct interactions between the different regions exchanged in the chimeras are most probably involved in forming KGF or bFGF binding sites.

• Steegmaier M et al.
• The E-selectin-ligand ESL-1 is a variant of a receptor for fibroblast growth factor.
• Nature. 1995; 373: 615-20
• Display abstract

• E-SELECTIN is an inducible cell-adhesion molecule on endothelial cells, which mediates the binding of neutrophils and functions as a Ca(2+)-dependent lectin. We have recently identified a 150K glycoprotein as the major ligand for E-selectin on myeloid cells, using a recombinant antibody-like form of mouse E-selectin as an affinity probe. Here we report the isolation of a mouse complementary DNA for this E-selectin ligand (ESL-1). The predicted amino-acid sequence of ESL-1 is 94% identical (over 1,078 amino acids) to the recently identified chicken cysteine-rich fibroblast growth-factor receptor, except for a unique 70-amino-acid aminoterminal domain of mature ESL-1. Fucosylation of ESL-1 is imperative for affinity isolation with E-selectin-IgG. A fucosylated, recombinant antibody-like form of ESL-1, but not of L-selectin, supports adhesion of E-selectin-transfected Chinese hamster ovary cells. Antibodies against ESL-1 block the binding of mouse myeloid cells to E-selectin. ESL-1, with a structure essentially identical to that of a receptor, thus functions as a cell adhesion ligand of E-selectin.

• Mathieu M et al.
• Receptor binding and mitogenic properties of mouse fibroblast growth factor 3. Modulation of response by heparin.
• J Biol Chem. 1995; 270: 24197-203
• Display abstract

• fgf3 has been implicated in the embryonic and fetal development of the mouse and as an oncogene in murine breast cancer. We describe a procedure to purify the product of the mouse fgf3 gene and show it to be a potent mitogen for some epithelial cell lines. Using a receptor binding competition assay, Fgf3 was shown to bind with high affinity to the IIIb isoforms of Fgf receptor (FgfR) 1 and FgfR2 (ID50 = approximately 0.8 nM) and with a lower affinity to the IIIc variant of FgfR2 (ID50 = approximately 9 nM). No competition for the binding of 125I-Fgf1 was observed for FgfR1 (IIIc), FgfR3 (IIIb and IIIc), or FgfR4. Mitogenicity assays using BaF3 cells containing individual Fgf receptors showed a pattern of response in agreement with the receptor binding results. A comparison of two mammary epithelial cell lines showed a marked difference of potency and dependence upon heparin in their response to mouse Fgf3, suggesting a complex interaction between the ligand and its low and high affinity receptors.

• Coutts JC, Gallagher JT
• Receptors for fibroblast growth factors.
• Immunol Cell Biol. 1995; 73: 584-9
• Display abstract

• The recent discovery of the involvement of heparan sulfate proteoglycans (HSPG) in the activation of fibroblast growth factor receptors (FGFR) has led to an intensification of study of this field. It appears that the HSPG act as low affinity receptors to which the fibroblast growth factors (FGF) must bind in order to successfully activate the high affinity FGFR. Heparan sulfate chains consisting of alternately arranged N-acetylated or N-sulfated glucosamine and uronic acid disaccharide regions, covalently attached to a core protein are found in two major families of cell surface HSPG, the syndecans and glypicans. A high affinity bFGF binding region has been isolated from fibroblast HS. There are four basic members of the FGFR family (FGFR 1-4), as well as a wealth of splice variants. The alternative forms of the basic receptors can have altered ligand binding or signalling qualities, depending on the region of the gene which is spliced. Investigations with null FGFR, incapable of signalling, have demonstrated the requirement for FGF in the organization of mammalian tissues and in embryonic patterning. Mutation of the FGFR genes has been recognized recently in human craniosynostoses where a single base pair mutation in the FGFR gene results in skeletal malformations specific to each syndrome. One suggestion is that the interaction of the mutant FGFR with the HSPG/FGF complex somehow contributes to the disease phenotype.

• Wong P, Hampton B, Szylobryt E, Gallagher AM, Jaye M, Burgess WH
• Analysis of putative heparin-binding domains of fibroblast growth factor-1. Using site-directed mutagenesis and peptide analogues.
• J Biol Chem. 1995; 270: 25805-11
• Display abstract

• The contribution of individual basic amino acids within three putative "consensus sequences" for heparin binding of fibroblast growth factor-1 have been examined by site-directed mutagenesis. The results indicate that a significant reduction in the apparent affinity of fibroblast growth factor-1 for heparin is only observed when basic residues in one of the three regions are mutated. Mutation in the other regions are without affect on heparin binding. The heparin binding properties of synthetic peptides based on the three "consensus sequences" paralleled the mutagenesis results. That is, synthetic peptides corresponding to regions of the protein that were affected by mutagenesis with respect to heparin binding exhibited a relatively high affinity for immobilized heparin, whereas those corresponding to regions of similar charge density that were unaffected by mutagenesis did not. In addition, amino acid substitution of a nonbasic residue in the heparin-binding peptide could abolish its heparin binding capacity. The heparin-binding peptide could antagonize the mitogenic activity of FGF-1, probably because of the heparin dependence of this activity. Together these data demonstrate that the heparin binding properties of fibroblast growth factor-1 are dictated by structural features more complex than clusters of basic amino acids. The results of these and other studies indicate that consensus motifs for heparin-binding require further definition. More importantly, the results provide a basis for the design of peptide-based inhibitors of FGF-1.

• Matsubara A
• [Studies on the mechanisms of action of fibroblast growth factors in stromal cells in hyperplastic human prostate]
• Nippon Hinyokika Gakkai Zasshi. 1995; 86: 1034-43
• Display abstract

• Stromal-epithelial cell interaction is very important for the development of human benign prostatic hyperplasia (BPH). Growth factors and their receptors in the prostate are thought to mediate the cell communication and play some roles in the development of BPH. Among many growth factors, fibroblast growth factor (FGF) family members have received the most intensive study, and mRNAs for acidic FGF (aFGF), basic FGF (bFGF) and keratinocyte growth factor (KGF) have been identified in rat or human prostate. However, synthesis sites and roles of them in stromal-epithelial cell interaction remain to be undefined. In the present study, to define the mechanisms for the regulation of prostatic cell growth by stromal cells in human BPH, we established the method for isolation and culture of epithelial cells as well as stromal cells from human BPH tissue. Using these primary cultured prostatic cells, we evaluated the effects of stromal cell conditioned medium (SCM) and stromal cell extract (SCE) on the growth of stromal cells and epithelial cells by [3H]-thymidine incorporation assay. We also examined the expression of mRNAs for aFGF, bFGF, KGF, FGF receptor 1 (FGFR1) and FGFR2 in epithelial and stromal cells using Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) analysis. As the results, both SCM and SCE stimulated the growth of stromal cells, and the growth promoting effects of them to stromal cells were completely suppressed by anti-bFGF neutralizing antibody, but not by anti-aFGF neutralizing antibody at all. SCM also stimulated the growth of epithelial cells. The growth promoting effect of it to epithelial cells was not completely suppressed by anti-bFGF neutralizing antibody, and not by anti-aFGF neutralizing antibody at all. Furthermore, RT-PCR analysis demonstrated the expression of mRNAs for bFGF, KGF and FGFR1 in stromal cells and that of FGFR2 in epithelial cells. These findings suggest that bFGF produced by stromal cells acts on stromal cells through FGFR1 by an autocrine mechanism, KGF produced by stromal cells acts on epithelial cells through FGFR2 by a paracrine manner in human BPH. These mechanisms for the regulation of cell growth by stromal cells were thought to contribute to the development of human BPH.

• Wanaka A
• Expression of heparin-binding growth factors and their receptors in neural development.
• Kaibogaku Zasshi. 1995; 70: 512-20

• Hall JG
• Information update on Achondroplasia.
• Pediatrics. 1995; 95: 620-620

• Wang F, Kan M, Xu J, Yan G, McKeehan WL
• Ligand-specific structural domains in the fibroblast growth factor receptor.
• J Biol Chem. 1995; 270: 10222-30
• Display abstract

• Two tandem immunoglobulin-like disulfide loops (Loops II and III) linked by a short connecting sequence in the ectodomain of the fibroblast growth factor receptor kinase compose the binding sites for glycosaminoglycan and fibroblast growth factor (FGF) ligands. Alternate splicing of exons IIIb and IIIc coding for the COOH-terminal half of Loop III confers high affinity for FGF-7 or FGF-2, respectively, on the fibroblast growth factor receptor ectodomain without effect on the binding of FGF-1. Here we show that a 139-amino acid fragment composed of Loop II, the inter-Loop II/III sequence, and a short segment of the NH2 terminus of Loop III is sufficient and near the minimal requirement for binding of FGF-1, FGF-2, and FGF-7. Extension of the fragment by five additional highly conserved residues (SD(P/A)QP) within a distinct constitutive structural domain (fl1) in Loop III restricts the binding of FGF-7 without effect on FGF-1 and FGF-2. Since the presence of exon IIIc in the full-length ectodomain does not change this ligand binding profile, we suggest that alternately spliced exon IIIc plays no active role in binding of the three ligands. In contrast, exon IIIb actively abrogates the restriction on the binding of FGF-7 and concurrently lowers the affinity for FGF-2.

• Kinoshita N, Minshull J, Kirschner MW
• The identification of two novel ligands of the FGF receptor by a yeast screening method and their activity in Xenopus development.
• Cell. 1995; 83: 621-30
• Display abstract

• We have developed a functional screen in yeast to identify ligands for receptor tyrosine kinases. Using this method, we cloned two Xenopus genes that activate the fibroblast growth factor (FGF) receptor. These encode novel secreted proteins, designated FRL1 and FRL2, distantly related to the epidermal growth factor and angiogenin/ribonuclease families, respectively. Both genes activate the FGF receptor in Xenopus oocytes as well as in yeast. Overexpression induces mesoderm and neural-specific genes in Xenopus explants; induction is blocked by a dominant negative inhibitor of the FGF receptor. FRL1 is broadly expressed during gastrulation and neurulation, while FRL2 is expressed principally in the axial mesoderm and brain at later stages. Our results indicate that despite their lack of similarity with FGF, FRL1 and FRL2 are ligands for the FGF receptor that play distinct roles in development.

• Bateman A, Chothia C
• Outline structures for the extracellular domains of the fibroblast growth factor receptors.
• Nat Struct Biol. 1995; 2: 1068-74
• Display abstract

• Fibroblast growth factor receptors (FGFRs) have three extracellular domains that belong to the immunoglobulin superfamily. We have determined the outline structures for these domains on the basis of their homology to the I set molecule telokin. The outline structures describe the relative positions of residues in each domain; their major secondary structures, and the extent to which residues are accessible to the solvent. They also provide the basis of a coherent description of the change in recognition properties that occur when the IIIb and IIIc exons are switched and of the effects of mutations in FGFRs that cause genetic diseases.

• DeVore DL, Horvitz HR, Stern MJ
• An FGF receptor signaling pathway is required for the normal cell migrations of the sex myoblasts in C. elegans hermaphrodites.
• Cell. 1995; 83: 611-20
• Display abstract

• The sex myoblasts (SMs) in C. elegans hermaphrodites undergo anteriorly directed cell migrations that allow for the proper localization of the egg-laying muscles. These migrations are controlled in part by a signal emanating from gonadal cells that allows the SMs to be attracted to their precise final positions flanking the center of the gonad. Mutations in egl-15 alter the nature of the interaction between the gonad and the SMs, resulting in the posterior displacement of the SMs. Here we show that egl-15 encodes a receptor tyrosine kinase of the fibroblast growth factor receptor (FGFR) subfamily with multiple roles in development. Three genes were identified that behave genetically as activators or mediators of egl-15 activity. One of these genes, sem-5, encodes an adaptor molecule that transduces signals from a variety of receptor tyrosine kinases. Like egl-15 and sem-5, the other two genes may similarly act in FGFR signaling pathways in C. elegans.

• Salamanca-Gomez F
• [Bone dysplasia, craniosynostoses, and fibroblast growth factor receptors]
• Gac Med Mex. 1995; 131: 357-8

• Hollway GE, Phillips HA, Ades LC, Haan EA, Mulley JC
• Localization of craniosynostosis Adelaide type to 4p16.
• Hum Mol Genet. 1995; 4: 681-3
• Display abstract

• Craniosynostosis Adelaide type is a rare autosomal dominant syndrome associated with digital abnormalities. Linkage mapping was carried out excluding allelism to Saethre-Chotzen syndrome at 7p21, craniosynostosis Boston type at 5q34-q35, Jackson-Weiss and Crouzon syndromes at 10q24-q25 and Pfeiffer syndrome mapping near 8cen. Exclusion mapping was extended to the entire genome until linkage to chromosome 4 was detected. A maximum two-point lod score of 6.2 (theta = 0.0) was obtained with D4S412. The gene responsible for craniosynostosis Adelaide type was localized to 4p16, telomeric to D4S394. This region contains two plausible candidate genes, the MSX1 (HOX7) homeobox gene and the FGFR3 fibroblast growth factor receptor gene.

• Lappi DA
• Tumor targeting through fibroblast growth factor receptors.
• Semin Cancer Biol. 1995; 6: 279-88
• Display abstract

• Fibroblast growth factor tyrosine kinase receptors are encoded by four genes, but alternate splicing can result in more than 100 possible protein sequences. The receptors have widespread expression in the developing embryo, but the expression becomes more restricted in the adult. The ligand-receptor relationship is complex, due to the diversity of the receptors and the large number of possible ligands: there are now nine (and probably more) members of the fibroblast growth factor family. This complicated ligand-receptor relationship creates many options to target cell types through the use of individual ligands or receptor-specific monoclonal antibodies. In-vivo data demonstrate that FGF receptors are expressed on tumor cells and can be used to target tumors for growth inhibition. Given the complexity, it is possible that a unique targetable FGF receptor isoform can be found in one or more tumor types. Examples of the targeting of growth inhibition agents to tumors through FGF receptors are discussed.

• Muenke M
• Finding genes involved in human developmental disorders.
• Curr Opin Genet Dev. 1995; 5: 354-61
• Display abstract

• Recent advances in the human genome initiative have accelerated positional cloning efforts toward identification of a number of genes responsible for human developmental anomalies, particularly those involving the skeletal system. Genotype/phenotype comparison and functional analysis of these genes will further elucidate pathways of normal and abnormal human development of the skeletal and other organ systems.

• Myoken Y, Myoken Y, Okamoto T, Sato JD, Takada K
• Effect of fibroblast growth factor-1 on the three-dimensional growth and morphogenesis of human salivary gland epithelial cells embedded in collagen gels.
• In Vitro Cell Dev Biol Anim. 1995; 31: 84-6

• Kardami E, Liu L, Kishore S, Pasumarthi B, Doble BW, Cattini PA
• Regulation of basic fibroblast growth factor (bFGF) and FGF receptors in the heart.
• Ann N Y Acad Sci. 1995; 752: 353-69

• Park WJ, Bellus GA, Jabs EW
• Mutations in fibroblast growth factor receptors: phenotypic consequences during eukaryotic development.
• Am J Hum Genet. 1995; 57: 748-54

• Rousseau F et al.
• Stop codon FGFR3 mutations in thanatophoric dwarfism type 1.
• Nat Genet. 1995; 10: 11-2

• Hamaguchi A, Tooyama I, Yoshiki T, Kimura H
• Demonstration of fibroblast growth factor receptor-I in human prostate by polymerase chain reaction and immunohistochemistry.
• Prostate. 1995; 27: 141-7
• Display abstract

• The expression and localization of fibroblast growth factor receptor-1 were investigated in human prostatic tissues with or without benign hyperplasia. Using a polymerase chain reaction method, we were able to demonstrate that prostatic tissues with benign hyperplasia expressed a significantly higher level of fibroblast growth factor receptor-1 mRNA than normal prostatic tissues (P < 0.01 by Anova). Western blot analysis using an antiserum against the receptor gave 2 bands with molecular weights of about 140 kDa and 80 kDa; these correspond to the expected sizes of the long and secreted forms of the fibroblast growth factor receptor-1, respectively. An immunohistochemical study using the same antiserum further demonstrated that the immunoreactive staining occurred mainly in the basal cells of the glandular epithelium and occasionally in the stromal cells. These results suggest that fibroblast growth factors may influence, at least in part, the proliferation of the epithelial cells seen in benign hyperplasia of human prostate.

• Schulte-Merker S, Smith JC
• Mesoderm formation in response to Brachyury requires FGF signalling.
• Curr Biol. 1995; 5: 62-7
• Display abstract

• BACKGROUND: The Brachyury (T) gene is required for the formation of posterior mesoderm and for axial development in both mouse and zebrafish embryos. In these species, and in Xenopus, the gene is expressed transiently throughout the presumptive mesoderm, and transcripts then persiste in notochord and posterior tissues. In Xenopus embryos, expression of the Xenopus homologue of Brachyury, Xbra, can be induced in presumptive ectoderm by basic fibroblast growth factor (FGF) and activin; in the absence of functional FGF or activin signalling pathways, expression of the gene is severely reduced. Ectopic expression of Xbra in presumptive ectoderm causes mesoderm to be formed. As Brachyury and its homologues encode sequence-specific DNA-binding proteins, it is likely that each functions by directly activating downstream mesoderm-specific genes. RESULTS: We show that expression in Xenopus embryos of RNA encoding a dominant-negative FGF receptor inhibits the mesoderm-inducing activity of Xbra. We demonstrate that ectopic expression of Xbra activates transcription of the embryonic FGF gene, and that embryonic FGF can induce expression of Xbra. This suggests that the two genes are components of a regulatory loop. Consistent with this idea, dissociation of Xbra-expressing cells causes a dramatic and rapid reduction in levels of Xbra, but the reduction can be inhibited by addition of FGF. CONCLUSION: Formation of mesoderm tissue requires an intact FGF signalling pathway downstream of Brachyury. This requirement is due to a regulatory loop, in which Brachyury activates expression of a member of the FGF family, and FGF maintains expression of Brachyury.(ABSTRACT TRUNCATED AT 250 WORDS)

• Parker TG
• Molecular biology of myocardial hypertrophy and failure: gene expression and trophic signaling.
• New Horiz. 1995; 3: 288-300
• Display abstract

• Pressure-overload cardiac hypertrophy is associated with the re-expression of an ensemble of genes representative of embryonic myocardium, whose protein products modulate myocardial function. Regulation of cardiac-specific gene expression in end-stage myocardial disease in humans implies a pathophysiologic role for altered gene expression in the progression from compensatory hypertrophy to decompensated heart failure. The molecular signals that transduce load into a hypertrophic cardiac myocyte phenotype involve mechanical deformation and the local myocardial production of trophic factors, including angiotensin II, and transforming and fibroblast growth factors. Growth factors provoke a pattern of gene expression in cultured cardiac myocytes resembling that seen in pressure overload in vivo, in keeping with an autocrine or paracrine model of hypertrophy. Moreover, growth factor stimulation and pressure-overload hypertrophy share intracellular signaling pathways, including the activation of nuclear proteins encoded by cellular oncogenes. Elucidation of these signaling pathways may provide new therapeutic targets for the treatment of cardiac muscle disease that overcomes the limitations of currently available strategies.

• Pirvola U, Cao Y, Oellig C, Suoqiang Z, Pettersson RF, Ylikoski J
• The site of action of neuronal acidic fibroblast growth factor is the organ of Corti of the rat cochlea.
• Proc Natl Acad Sci U S A. 1995; 92: 9269-73
• Display abstract

• Here we show that the mature cochlear neurons are a rich source of acidic fibroblast growth factor (aFGF), which is expressed in the neuronal circuitry consisting of afferent and efferent innervation. The site of action of neuronal aFGF is likely to reside in the organ of Corti, where one of the four known FGF receptor (FGFR) tyrosine kinases--namely, FGFR-3 mRNA--is expressed. Following acoustic overstimulation, known to cause damage to the organ of Corti, a rapid up-regulation of FGFR-3 is evident in this sensory epithelium, at both mRNA and protein levels. The present results provide in vivo evidence for aFGF being a sensory neuron-derived, anterogradely transported factor that may exert trophic effects on a peripheral target tissue. In this sensory system, aFGF, rather than being a neurotrophic factor, seems to promote maintenance of the integrity of the organ of Corti. In addition, aFGF, released from the traumatized nerve endings, may be one of the first signals initiating protective recovery and repair processes following damaging auditory stimuli.

• Gray TE, Eisenstein M, Shimon T, Givol D, Yayon A
• Molecular modeling based mutagenesis defines ligand binding and specificity determining regions of fibroblast growth factor receptors.
• Biochemistry. 1995; 34: 10325-33
• Display abstract

• The fibroblast growth factor receptor 2 (FGFR2) and the keratinocyte growth factor receptor (KGFR) have different ligand binding specificities despite differing only in the second half of their immunoglobulin-like (Ig-like) domain III. Three-dimensional model structures were generated for domain III on the basis of variable (V) Ig domains. The region that differs between the two receptors is predicted to include two loops: one connects beta-strands F-G and is analogous to the complementarity determining region 3 (CDR3) of immunoglobulins; the other connects beta-strands D-E. These regions were targeted for mutagenesis. Single mutations in the F-G loop were found to only slightly alter ligand binding, whereas a double mutant, KGFR Y345-->S,Q348-->I, acquired significant affinity for bFGF. Notably, the affinity of this double mutant KGFR for KGF and aFGF was essentially unaltered. A mutant FGFR2, in which the D-E beta-hairpin (T319TDKEI) is replaced with the KGFR D-E beta-hairpin (S319SNA), has 9-fold reduced affinity for bFGF. These results demonstrate that the F-G or CDR3 analogous loop in FGFRs plays a key role in determining ligand binding and specificity. In addition, however, the protein loop connecting beta-strands D and E may also be involved in ligand binding. Several point mutations in FGFR2, shown recently to give rise to multiple inherited skeletal defects, are localized according to our models to the F-G or D-E loops of domain III. Our results strongly suggest that these naturally occurring mutations specifically alter ligand binding by FGFR2.

• Schmidt KF, Billek M, Pietruck C, Noll GN, Goureau O, Courtois Y
• Fibroblast growth factors alter light responses and dark voltage in retinal rods of the frog (Rana temporaria).
• Neurosci Lett. 1995; 191: 177-80
• Display abstract

• Fibroblast growth factors (FGF-1 and FGF-2) were applied intracellularly via whole-cell patch-clamp electrodes while the membrane voltage was recorded simultaneously. During recording the exchange of substances by diffusion between cytosol and pipette medium affects the cell's function. Under control conditions, the loss of nucleotides is reflected by a slow hyperpolarization of the dark voltage and prolongated light responses. Addition of FGF-1 and FGF-2 to the pipette medium accelerated the time course of the hyperpolarization and intensified the prolongation of the light responses. The depolarization of photoreceptor cells after intracellular application of the nitric oxide (NO)-synthase cofactor nicotinamide adenine dinucleotide phosphate (NADPH) and the stabilization of light response recovery by L-arginine is abolished by FGF-2. FGF-2 was ineffective when it was applied together with the calcium chelator ethylene glycol-bis(2-aminoethylether)tetraacetate (EGTA). The results indicate a possible role of FGF in the regulation of NO and calcium in photoreceptor cells and may explain protective effects of FGF in degenerative processes of photoreceptor cells.

• Prochiantz A, Theodore L
• Nuclear/growth factors.
• Bioessays. 1995; 17: 39-44
• Display abstract

• The now classical model for cell-cell communication espouses that information travels between cells in the form of molecules that bind specific cell-surface receptors and trigger signal-transducing mechanisms that eventually lead to transcriptional modifications. Here we gather the available information suggesting that some growth factors may also act by interfering directly with gene transcription, following their internalization and nuclear translocation. Among these factors are bona fide growth factors such as Fibroblast Growth Factor-1 and -2 and Schwannoma Derived Growth Factor, for which internalization and nuclear translocation have been demonstrated. Conversely, we propose that some isoforms of nuclear factors of the homeoprotein family could pass from cell to cell. The implications of the model are presented in the context of the specificity of cellular interactions.

• Kuzis K, Reed S, Cherry NJ, Woodward WR, Eckenstein FP
• Developmental time course of acidic and basic fibroblast growth factors' expression in distinct cellular populations of the rat central nervous system.
• J Comp Neurol. 1995; 358: 142-53
• Display abstract

• Acidic and basic fibroblast growth factors (aFGF and bFGF, respectively) are expressed in high levels in adult central nervous system (CNS). We report the time course of developmental appearance and distribution of these factors and of two FGF receptors, FGFR-1 and FGFR-2, in the CNS of rats ranging in age from embryonic day 16 to adult. Immunohistochemical analysis showed that sensory neurons in the midbrain were the first cells to contain detectable aFGF immunoreactivity at embryonic day 18. The next cell group to contain aFGF were motor neurons, which were found to be aFGF-positive at the day of birth. A number of other subcortical neuronal populations were observed to contain aFGF immunoreactivity after postnatal day 7. Adult levels and distribution patterns of aFGF were reached in all CNS areas by postnatal day 28. Basic FGF immunoreactivity was observed at postnatal day 0 in neurons in the CA2 subfield of hippocampus. Astrocytes contained detectable bFGF immunoreactivity, starting at postnatal day 7. Adult levels and patterns of distribution of bFGF were reached in all CNS areas by postnatal day 28. These immunohistochemical observations were confirmed by using bioassay and Western blot techniques. FGFR-1 and FGFR-2 mRNA were expressed in significant levels in all CNS areas at all time points analyzed. The observation that aFGF and bFGF appear in specific and distinct cellular populations at relatively late developmental times suggests that these FGFs may be involved in specific mechanisms of CNS maturation, maintenance, and repair.

• Palcy S, Chopin DK, Caruelle D, Ravery V, Barritault D, Caruelle JP
• Fibroblast growth factors and their specific binding sites in normal urothelium and transitional cell carcinoma (TCC) of the human bladder.
• Cell Mol Biol (Noisy-le-grand). 1995; 41: 565-75
• Display abstract

• In order to further investigate the involvement of FGF1 and FGF2 in the transitional cell carcinoma of the bladder (TCC), we compared the expression of FGF1, FGF2 and their membrane binding sites by biochemical and immunological methods in both human normal urothelium and transitional cell carcinoma of the bladder. Using a specific enzyme immunoassay (EIA), we observed that neoplastic tissue exhibited a lower FGF2 content than normal tissue. FGF1 and FGF2 Western-blot analysis revealed in both types of tissues a major FGF1 form with a low molecular weight (14 kDa) and a single FGF2 form (21 kDa). Immunohistological studies showed that FGF1 was specifically located in normal or transformed urothelial cells, while FGF2 was associated with bladder stroma. Binding analysis in tissue membrane preparations revealed a dramatic drop in both high and low affinity binding sites for FGF2 in TCC bladder in comparison to normal bladder. FGF2 cross-linking experiments illustrated a qualitative modification of FGF2 binding complexes in tumors. These data suggest that invasive bladder carcinoma is associated with modifications of FGF1 and FGF2 tissue levels and alterations in the characteristics of the FGF2 membrane binding sites.

• Francomano CA
• The genetic basis of dwarfism.
• N Engl J Med. 1995; 332: 58-9

• Hastbacka J
• The molecular basis of hereditary skeletal disorders.
• Ann Med. 1995; 27: 433-4

• Kiefer P, Dickson C
• Nucleolar association of fibroblast growth factor 3 via specific sequence motifs has inhibitory effects on cell growth.
• Mol Cell Biol. 1995; 15: 4364-74
• Display abstract

• The dual subcellular fate of fibroblast growth factor 3 (FGF3) is determined by the competing effects of amino-terminal signals for nuclear localization and secretion (P. Kiefer, P. Acland, D. Pappin, G. Peters, and C. Dickson, EMBO J. 13:4126-4136, 1994). Mutation analysis has implicated additional basic domains in the carboxy-terminal region of the protein as necessary for nuclear uptake and the association of FGF3 with the nucleoli. Immunogold electron microscopy shows that FGF3 is predominantly within the dense fibrillar component of the nucleolus. A form of FGF3 that localizes exclusively in the nucleus and nucleolus was generated by removing signals for secretion, and expression of this nonsecreted FGF3 in a mammary epithelial cell line resulted in slowly growing colonies of enlarged cells. Thus, nuclear import and nucleolar association of FGF3 are determined by the concerted interaction of several distinct motifs, and the exclusive production of the nuclear isoform can inhibit DNA synthesis and cell proliferation.

• Story MT
• Regulation of prostate growth by fibroblast growth factors.
• World J Urol. 1995; 13: 297-305
• Display abstract

• Growth of the prostate is controlled by androgen. However, there is information indicating that androgen may not act directly, but may act indirectly through polypeptide growth factors, to control prostate growth. This review will focus on the involvement of members of the fibroblast growth factor (FGF) family in this process. The properties of FGFs and FGF-receptors are described that implicate these molecules in growth control. Information is provided that prostate stromal cells synthesize FGF2 and FGF7. FGF2 is a potent mitogen for stromal cells; whereas, FGF7 is exclusively a mitogen for epithelial cells. Transforming growth factor beta (TGF beta), also produced by prostate cells, inhibit cell growth. This suggests that prostate growth is controlled by autocrine and paracrine mechanisms. Evidence is presented that altered FGF expression accompanies benign prostatic hyperplasia and prostate cancer. A model is proposed whereby androgen regulates TGF beta, influencing FGF2 and FGF7 expression, and in turn regulating growth of the prostatic stroma and epithelium. An imbalance in the influence of these growth factors may contribute to prostate disease.

• Brew EC, Mitchell MB, Harken AH
• Fibroblast growth factors in operative wound healing.
• J Am Coll Surg. 1995; 180: 499-504

• Stieber A, Mourelatos Z, Chen YJ, Le Douarin N, Gonatas NK
• MG160, a membrane protein of the Golgi apparatus which is homologous to a fibroblast growth factor receptor and to a ligand for E-selectin, is found only in the Golgi apparatus and appears early in chicken embryo development.
• Exp Cell Res. 1995; 219: 562-70
• Display abstract

• While over 20 intrinsic proteins of the Golgi apparatus have been identified and sequenced, there is no information on their developmental history, i.e., whether all Golgi proteins are expressed simultaneously or whether there is a hierarchical or stage-specific order of their expression during embryonic development. In this study we have examined the emergence and distribution of MG160 during the development of chicken embryos. MG160 is a conserved membrane sialoglycoprotein of the Golgi apparatus of most cells displaying over 90% amino acid sequence identities with two apparently unrelated molecules, namely CFR, a chicken fibroblast growth factor receptor, and ESL-1, a ligand for E-selectin (Gonatas et al., J. Biol. Chem. 1989, 264, 646-653; Burrus and Olwin, J. Biol. Chem. 1989, 264, 18647-18653; Burrus et al., Mol. Cell Biol. 1992, 12, 5600-5609; Gonatas et al., J. Cell Sci. 108, 457-467; Steegmaier et al., Nature 1995, 373, 615-620). This study was carried out by in situ hybridization, using a 56-mer antisense probe for the chicken homologue of MG160 which differs only by four bases from the corresponding segment of the rat cDNA and by immunocytochemistry and Western blotting using a polyclonal antiserum against MG160. The protein was ubiquitously and exclusively localized in the Golgi apparatus and appeared early in development within the ectoblast and primitive endoblast prior to the formation of the primitive streak. At 2 to 3 days, MG160 was particularly prominent in the notochord, neural tube, somites, and cartilage cells. In organs with central lumens, such as the neural tube, the Golgi apparatus, visualized by immunostaining for MG160, was elongated and it was located at the apical pole of cells. In 6-day-old embryos, the ongoing physiologic degeneration of the notochord was accompanied by fragmentation of the immunostained Golgi apparatus and decreased labeling of the mRNA for MG160. In order to gain information on possible interactions between MG160 and basic fibroblast growth factor (bFGF), the localization of both molecules was studied by immunocytochemistry in 3-day-old chicken embryos. While MG160 was ubiquitous in the Golgi apparatus of all cells and tissues, endogenous bFGF was no detected, while exogenous bFGF bound only to basement membranes. These results indicate that MG160 is a primordial protein of the Golgi apparatus and are consistent with the hypothesis that the binding of MG160 to fibroblast growth factors and E-selectin is not related to the still unknown principal function of MG160 in the Golgi apparatus.

• Saffell JL, Williams EJ, Doherty P, Walsh FW
• Axonal growth mediated by cell adhesion molecules requires activation of fibroblast growth factor receptors.
• Biochem Soc Trans. 1995; 23: 469-70

• Tabin C
• The initiation of the limb bud: growth factors, Hox genes, and retinoids.
• Cell. 1995; 80: 671-4

• Cuevas P, Carceller F, Gimenez-Gallego G
• Fibroblast growth factor and cerebral ischaemia.
• Neurol Res. 1994; 16: 181-3
• Display abstract

• Although the cellular events that accompany brain injury have been well characterized, the factors that mediate growth and repair in the central nervous system (CNS) are only beginning to be identified. One such is fibroblast growth factor. The fibroblast growth factor (FGF) family of proteins includes, at this moment, seven structurally related polypeptides. Acidic and basic FGF (aFGF and bFGF) were the first described members of the family, and are the best characterized. Both proteins bind with high affinity to the same kind of receptor at the plasma membrane through which they induce a whole series of intracellular events that mediate their physiological functions. FGF, which displays pleiotrophic biological activities including mitogenesis, chemotaxis, and induction of differentiation is a potent angiogenic factor in vivo, tested in assays as different as the rabbit cornea, the chick chorioalantoic membrane (CAM), the rat common carotid artery, and the rat brain.

• Li Y, Basilico C, Mansukhani A
• Cell transformation by fibroblast growth factors can be suppressed by truncated fibroblast growth factor receptors.
• Mol Cell Biol. 1994; 14: 7660-9
• Display abstract

• Ligand-induced dimerization and transphosphorylation are thought to be important events by which receptor tyrosine kinases generate cellular signals. We have investigated the ability of signalling-defective, truncated fibroblast growth factor (FGF) receptors (FGFR-1 and FGFR-2) to block the FGF response in cells that express both types of endogenous FGF receptors. When these dominant negative receptors are expressed in NIH 3T3 cells transformed by the secreted FGF-4, the transformed properties of the cells can be reverted to various degrees, with better reversion phenotype correlating with higher levels of truncated receptor expression. Furthermore, truncated FGFR-2 is significantly more efficient at producing reversion than FGFR-1, indicating that FGF-4 preferentially utilizes the FGFR-2 signalling pathway. NIH 3T3 clones expressing these truncated receptors are more resistant to FGF-induced mitogenesis and also exhibit reduced tyrosine phosphorylation upon treatment with FGF. The block in FGF-signalling, however, can be overcome by the addition of excess growth factor. The truncated receptors have binding affinities that are four- to eightfold lower than those of wild-type receptors, as measured by Scatchard analysis. We also observed a partial specificity in the responses of truncated-receptor-expressing clones to FGF-2 or FGF-4. Our results suggest that the block to signal transduction produced by kinase-negative FGF receptors is achieved through a combination of dominant negative effects and competition for growth factor binding with functional receptors.

• Wang G, Thomas KA
• Purification and characterization of a functional soluble fibroblast growth factor receptor 1.
• Biochem Biophys Res Commun. 1994; 203: 1781-8
• Display abstract

• Naturally encoded human soluble fibroblast growth factor receptor 1 was cloned, abundantly expressed by recombinant baculovirus-infected insect cells and purified to apparent homogeneity. Pure soluble receptor bound as monomeric and dimeric complexes and inhibited the mitogenic activity of basic and acidic fibroblast growth factors.

• Harland RM
• Neural induction in Xenopus.
• Curr Opin Genet Dev. 1994; 4: 543-9
• Display abstract

• Although induction of neural tissue in vertebrates has been recognized since the experiments of Spemann and Mangold seventy years ago, only recently has the phenomenon been put onto a molecular footing. Three molecules, noggin, follistatin and fibroblast growth factor, have been shown to have neuralizing activity in various assays. These assays for neural induction and the molecular mechanisms for the action of neural inducers are reviewed here.

• Sorokin A, Mohammadi M, Huang J, Schlessinger J
• Internalization of fibroblast growth factor receptor is inhibited by a point mutation at tyrosine 766.
• J Biol Chem. 1994; 269: 17056-61
• Display abstract

• Binding of fibroblast growth factor (FGF) to the fibroblast growth factor receptor leads to autophosphorylation of the receptor on several tyrosine residues. Wild-type FGF receptor 1 (flg) and a mutated receptor (Y766F), in which an autophosphorylation site (Tyr-766) was mutated to phenylalanine, were expressed in rat myoblasts and in hematopoietic Ba/F3 cells. It was found that the point mutation at Tyr-766 resulted in a decrease in FGF receptor internalization, as well as a reduction in both ligand-induced FGF receptor down-regulation and degradation. It has been shown previously that phosphorylation of Tyr-766 is essential for interaction with phospholipase C gamma and that the Y766F FGF receptor mutant is unable to stimulate phosphatidylinositol hydrolysis and Ca2+ release from internal stores. The results presented in this report indicate that Tyr-766 is also essential for cellular trafficking of FGF receptor.

• Kimura H, Tooyama I, McGeer PL
• Acidic FGF expression in the surroundings of senile plaques.
• Tohoku J Exp Med. 1994; 174: 279-93
• Display abstract

• Immunohistochemical examination of postmortem brain tissue of Alzheimer's disease revealed that acidic fibroblast growth factor (aFGF) was specifically expressed in a subpopulation of reactive astrocytes which were congregated at the margin of the senile plaque. Double immunostaining indicated that such upregulation of aFGF expression might be related to the presence of reactive microglia rather than beta-amyloid protein deposits. Although, on the other hand, immunohistochemical staining for fibroblast growth factor receptor-1 occurred in some cortical neurons of Alzheimer's disease, the staining pattern did not differ from that in age-matched controls. Possible significance of aFGF-positive astrocytes in the surroundings of the senile plaque will be discussed in relation to receptor mediated or non-mediated mechanisms.

• Olwin BB, Hannon K, Kudla AJ
• Are fibroblast growth factors regulators of myogenesis in vivo?
• Prog Growth Factor Res. 1994; 5: 145-58
• Display abstract

• Recent advances in understanding of skeletal muscle differentiation implicate fibroblast growth factors (FGFs) as regulators of myogenesis; however, the identity and actions of factors that repress myogenesis in vivo remain to be established. This review will focus on the fibroblast growth factor family and the evidence for its role in regulating myogenesis in culture and in vivo.

• Eckenstein FP
• Fibroblast growth factors in the nervous system.
• J Neurobiol. 1994; 25: 1467-80
• Display abstract

• Fibroblast growth factors (FGFs) exhibit widespread mitogenic and neurotrophic activities. Nine members of the family are currently known, and FGF-1 and FGF-2 are present in relatively high levels in CNS. FGF-1 is expressed by a subset of neuronal populations, while FGF-2 is expressed by astrocytes. FGF-1 and FGF-2 lack signal peptides and appear to be present mainly in intracellular compartments. This suggests that the factors may act as initiators of a repair response after injury. Support for this notion comes from observations that FGF-1 and FGF-2 levels are low during critical phases of development, but high in the adult CNS. A family of transmembrane tyrosine kinase receptors (FGFRs) mediates the effects of FGFs. Four different genes coding for FGF receptors are currently known, three of which are expressed in cell type-specific patterns in the CNS. The main receptor variants present in this tissue, however, can by themselves not distinguish between FGF-1 and FGF-2. Additional selectivity may be established by interaction of the FGFs and their receptors with select heparan proteoglycans (HSPGs). Therefore, the precise physiological role of FGFs is determined by the combination of cell type-specific patterns of expression of FGFs, FGFRs and HSPGs together with the mechanisms that regulate the extracellular availability of FGFs.

• Hanneken A, Ying W, Ling N, Baird A
• Identification of soluble forms of the fibroblast growth factor receptor in blood.
• Proc Natl Acad Sci U S A. 1994; 91: 9170-4
• Display abstract

• We have purified three acidic (FGF-1) and basic (FGF-2) fibroblast growth factor binding proteins (FGF-BP1, FGF-BP2, and FGF-BP3) from human plasma and calf serum and demonstrate the presence of these circulating FGF-BPs in blood. Each are truncated forms of the high-affinity FGF receptor (FGFR-1). FGF-BP1 and FGF-BP2 have estimated molecular masses of 70-85 kDa and 55-60 kDa, respectively, and are detected by using 125I-labeled FGF-2 ligand blotting. Immunoblotting with four distinct antibodies to FGFR-1 reveals that FGF-BP1 and FGF-BP2 are immunologically and biochemically related to the extracellular domain of FGFR-1. Reverse-phase HPLC chromatography resolves FGF-BP2 into two proteins with estimated molecular masses of 55 kDa and 60 kDa. Protein sequencing of the amino terminus of FGF-BP2 and FGF-BP3 reveals identity with the extracellular domain of the two-IgG-loop form of human FGFR-1. The FGF-BPs do not require heparin to bind FGF-2 on affinity columns, but heparin does enhance their recovery from blood. These FGF-BPs may play an important physiological role in regulating the biological activity of FGF and the other members of the FGF family of growth factors.

• Siegall CB, Gawlak SL, Chace DF, Merwin JR, Pastan I
• In vivo activities of acidic fibroblast growth factor-Pseudomonas exotoxin fusion proteins.
• Bioconjug Chem. 1994; 5: 77-83
• Display abstract

• Fibroblast growth factor receptors are highly expressed in a variety of cancer cells and activated vasculature. Using chimeric toxins targeted to cell-surface a FGF receptors, we have demonstrated specific cytotoxic activity to these cell types. These molecules, aFGF-PE40 and aFGF-PE40 KDEL, are fusion proteins containing acidic FGF and either a 40- or a 66-kDa binding defective form of Pseudomonas exotoxin, respectively. Both aFGF-toxin fusion proteins were able to inhibit protein synthesis in vitro in a variety of carcinoma cell lines. The half-life of aFGF-PE40 in serum was found to be 41 min when coadministered with heparin. Administration of aFGF-PE40 or aFGF-PE4E KDEL with heparin inhibits the growth of established KB and preestablished A431 epidermoid carcinoma xenografts in athymic mice. The antitumor activities of the two aFGF-toxin fusion proteins were equivalent against the KB tumor xenografts. While we were able to slow the growth of the KB tumor xenografts, we were unable to cause tumor regressions. Histochemical analysis of treated versus untreated tumor tissue revealed a difference in tumor size but not of vascularity. We conclude that aFGF-PE40 and aFGF-PE4E KDEL have in vivo antitumor activity that targets the tumor cell mass rather than vascular structures in mice xenografted with human epidermoid carcinoma.

• Friess H, Korc M, Buchler MW
• [Molecular biology of pancreatic cancer: overexpression of fibroblast growth factors]
• Chirurg. 1994; 65: 604-10
• Display abstract

• In the present study, the expression of a acidic fibroblast growth factor (aFGF), basic fibroblast growth factor (bFGF) and fibroblast growth factor receptor-1 were analyzed in 60 pancreatic cancer samples using Northern blot analysis, immunoblotting, in situ hybridization and immunohistochemical techniques. aFGF, bFGF and FGFR-1 were present in 63%, 55% and 52% of the tumor samples, respectively. Twenty-seven of the 60 pancreatic cancer tissue samples exhibited coexpression of aFGF and bFGF. In contrast, 14 of the tumor samples did not show immunoreactivity for aFGF or bFGF in the tumor cells. The expression of aFGF in the tumor cells had no influence on the postoperative survival period, whereas patients whose tumors were positive for bFGF and/or FGF-receptor-1 had significantly shorter postoperative survival periods. Our results suggest that bFGF and FGFR-1 may play a role in the growth behavior of pancreatic cancer and may contribute to tumor aggressiveness.

• Gallagher JT
• Heparan sulphates as membrane receptors for the fibroblast growth factors.
• Eur J Clin Chem Clin Biochem. 1994; 32: 239-47
• Display abstract

• The complex polymeric structure of heparan sulphate contains intra-chain sequences of sulphated sugar residues that define active sites for the binding and activation of growth factors. The molecular mechanisms of recognition and activation are slowly being revealed at least in the case of the interaction of heparan sulphate with basic fibroblast growth factor. Current data indicate that relatively long but specific binding sequences in heparan sulphate may induce a conformational change in basic fibroblast growth factor exposing a site on the protein that is recognised by signal transducing receptors. Heparan sulphate may also subserve functions of dimerisation of basic fibroblast growth factor and facilitation of receptor transfer by a secondary interaction with the receptor itself. Various models for heparan sulphate mediated induction of mitogenesis by basic fibroblast growth factor have been proposed and there are suggestions that the core protein of plasma membrane heparan sulphate-proteoglycans may participate in the cell signalling process. The vital importance of heparan sulphate in controlling growth factor activities has opened up a new chapter in proteoglycan research and has brought proteoglycans into the mainstream of cell biology. Further investigation of their mode of action is likely to reveal new information on the control of cell growth and development in both embryonic and adult tissues and may suggest novel methods of controlling diseases such as cancer, atherosclerosis or fibrosis that are driven by abnormal expression of growth factors or their receptors.

• Goncharova VP
• [Fibroblast growth factors (a short review)]
• Fiziol Zh Im I M Sechenova. 1994; 80: 163-74
• Display abstract

• Some features of structure and expression of bFGF gene and FGF receptors genes, unique secretion property of bFGF, as well as FGF's interactions with heparin-containing components of the cell and extracellular matrix are briefly observed. These special features may underlie the multifunctionality of members of ubiquitously distributed fibroblast growth factor family which recently have been filled up with two new members: androgen-induced growth factor--FGF-8 (Tanaka A., Miyamoto K., Minamino N. at al., 1992) and glia-activating factor--FGF-9 (Naruo K., Seco C., Kuroshima K. at al. 1993).

• Jouanneau J, Moens G, Bourgeois Y, Poupon MF, Thiery JP
• A minority of carcinoma cells producing acidic fibroblast growth factor induces a community effect for tumor progression.
• Proc Natl Acad Sci U S A. 1994; 91: 286-90
• Display abstract

• It is generally accepted that primary tumors become heterogeneous as a consequence of tumor-cell genetic instability. Clonal dominance has been shown to occur in some experimental models allowing a subpopulation of cells to overgrow the primary heterogeneous tumor and to metastasize. Alternatively, interactions among coexisting tumor subpopulations may contribute to the emergence of a malignant invasive primary solid tumor. We asked the question whether emergence of carcinoma cells producing a growth/dissociating factor within a tumor cell population may be a determinant for tumor progression and for clonal dominance. To mimic such a situation, we have investigated the impact of tumor subpopulation heterogeneity in an in vivo model in which mixtures of carcinoma cells that differ in their ability to produce acidic fibroblast growth factor are injected into nude mice. Our data indicate that a growth-factor-producing cell subpopulation can confer increased tumorigenicity to an entire cell population and subsequently elicit a shorter delay for appearance of metastasis. A community effect via cell interactions may account for a heterogeneous tumor cell population rather than clonal dominance during progression of certain tumor types.

• Slack JM, Isaacs HV
• The role of fibroblast growth factors in early Xenopus development.
• Biochem Soc Trans. 1994; 22: 585-9

• Spivak-Kroizman T, Mohammadi M, Hu P, Jaye M, Schlessinger J, Lax I
• Point mutation in the fibroblast growth factor receptor eliminates phosphatidylinositol hydrolysis without affecting neuronal differentiation of PC12 cells.
• J Biol Chem. 1994; 269: 14419-23
• Display abstract

• Fibroblast growth factors (FGF) stimulate growth arrest and differentiation in rat pheochromocytoma PC12 cells. We examined the role of phosphatidylinositol (PI) hydrolysis in FGF-induced differentiation of PC12 cells by exploring the biological and biochemical activity of a mutant FGF receptor 1 (flg) defective in stimulation of PI hydrolysis. We show that point mutation at Tyr-766 (Y766F) of the FGF receptor prevents tyrosine phosphorylation of phospholipase C gamma and eliminates acidic FGF (aFGF)-induced stimulation of PI hydrolysis in PC12 cells. Treatment of PC12 cells expressing either wild-type or the Y766F mutant with aFGF led to tyrosine phosphorylation of Shc, the association of Shc with GRB2, a shift in the electrophoretic mobility of the Ras guanine nucleotide-releasing factor, Sos (son of sevenless), and enhancement in mitogen-activated protein kinase phosphorylation. Moreover, stimulation with aFGF led to a typical neurite outgrowth of PC12 cells expressing either wild-type or the Y766F FGF receptor mutant. These experiments indicate that PI hydrolysis is not essential for FGF-induced neuronal differentiation of PC12 cells. Moreover, the aFGF-induced Ras signaling pathway, which is essential for PC12 cell differentiation, is not affected by elimination of PI hydrolysis.

• Pantoliano MW et al.
• Multivalent ligand-receptor binding interactions in the fibroblast growth factor system produce a cooperative growth factor and heparin mechanism for receptor dimerization.
• Biochemistry. 1994; 33: 10229-48
• Display abstract

• The binding interactions for the three primary reactants of the fibroblast growth factor (FGF) system, basic FGF (bFGF), an FGF receptor, FGFR1, and the cofactor heparin/heparan sulfate (HS), were explored by isothermal titrating calorimetry, ultracentrifugation, and molecular modeling. The binding reactions were first dissected into three binary reactions: (1) FGFR1 + bFGF<==>FGFR1/bFGF, K1 = 41 (+/- 12) nM; (2) FGFR1 + HS<==>FGFR1/HS, K2 = 104 (+/- 17) microM; and (3) bFGF + HS<==>bFGF/HS, K3 = 470 (+/- 20) nM, where HS = low MW heparin, approximately 3 kDa. The first, binding of bFGF to FGFR1 in the absence of HS, was found to be a simple binary binding reaction that is enthalpy dominated and characterized by a single equilibrium constant, K1. The conditional reactions of bFGF and FGFR1 in the presence of heparin were then examined under conditions that saturate only the bFGF heparin site (1.5 equiv of HS/bFGF) or saturate the HS binding sites of both bFGF and FGFR1 (1.0 mM HS). Both 3-and 5-kDa low MW heparins increased the affinity for FGFR1 binding to bFGF by approximately 10-fold (Kd = 4.9 +/- 2.0 nM), relative to the reaction with no HS. In addition, HS, at a minimum of 1.5 equiv/bFGF, induced a second FGFR1 molecule to bind to another lower affinity secondary site on bFGF (K4 = 1.9 +/- 0.7 microM) in an entropy-dominated reaction to yield a quaternary complex containing two FGFR1, one bFGF, and at least one HS. Molecular weight estimates by analytical ultracentrifugation of such fully bound complexes were consistent with this proposed composition. To understand these binding reactions in terms of structural components of FGFR1, a three-dimensional model of FGFR1 was constructed using segment match modeling. Electrostatic potential calculations confirmed that an elongated cluster, approximately 15 x 35 A, of nine cationic residues focused positive potential (+2kBT) to the solvent-exposed beta-sheet A, B, E, C' surface of the D(II) domain model, strongly implicating this locus as the HS binding region of FGFR1. Structural models for HS binding to FGFR1, and HS binding to bFGF, were built individually and then assembled to juxtapose adjacent binding sites for receptor and HS on bFGF, against matching proposed growth factor and HS binding sites on FGFR1. The calorimetric binding results and the molecular modeling exercises suggest that bFGF and HS participate in a concerted bridge mechanism for the dimerization of FGFR1 in vitro and presumably for mitogenic signal transduction in vivo.(ABSTRACT TRUNCATED AT 400 WORDS)

• Fernig DG, Gallagher JT
• Fibroblast growth factors and their receptors: an information network controlling tissue growth, morphogenesis and repair.
• Prog Growth Factor Res. 1994; 5: 353-77
• Display abstract

• The stimulation of cellular metabolism by the nine fibroblast growth factors (FGFs) is mediated by a dual-receptor system. This comprises a family of four receptor tyrosine kinases (FGFR) and heparan sulphate proteoglycans (HSPG). The stimulation of cell division by FGFs has an obligate requirement for both partners of the dual-receptor system. The binding of the nine FGFs to the FGFRs is marked by a pattern of overlapping specificity despite alternative splicing events generating a large number of FGFR proteins. Thus many of the FGFR isoforms bind several FGFs. It is likely that each FGF requires a different pattern of sulphation within the heparan sulphate chains for binding. Therefore, the HSPG receptors may provide additional specificity, allowing a cell to fine tune its response to the FGFs present in the extracellular milieu. The HSPG receptors also control the availability of FGFs and hence regulate the transport of FGFs within a tissue. FGF-stimulated cell division would appear to have a mandatory requirement for the FGFs to be translocated to the nucleus via the cytosol after interacting with the dual-receptor system. The consequences of the potential direct action of FGFs in stimulating cell division are examined in the light of current models of signal transduction.

• Matsuo A et al.
• Immunohistochemical localization in the rat brain of an epitope corresponding to the fibroblast growth factor receptor-1.
• Neuroscience. 1994; 60: 49-66
• Display abstract

• The localization of fibroblast growth factor receptor-1 was investigated in rat brain by immunohistochemistry using a polyclonal antibody against an acidic peptide sequence of chicken fibroblast growth factor receptor-1. For raising the antisera in rabbits, we synthesized the oligopeptide EDDDDEDDSSSEEKEAD which is a highly acidic region of chicken fibroblast growth factor receptor-1. The oligopeptide was used as a haptenic antigen by conjugating with poly-L-glutamate as a carrier protein. On immunospot assay, the best antiserum was capable of detecting 15.7 pmols of both the chicken and its analogous human oligopeptides but failed to react even with up to 1 nmol of poly-L-glutamate. When rat brain homogenate was examined by Western blots, the antiserum revealed two bands with molecular weights of 145,000 and 75,000 corresponding to known sizes of the membrane-bound and secreted forms of the rat receptor, respectively. Immunohistochemistry in rat brain demonstrated that putative fibroblast growth factor receptor-1 immunoreactivity sites were present mainly in neurons but also in tanycytes and ependymal cells. Positive neurons were distributed widely in various brain regions, but were particularly abundant in such regions as the lateral hypothalamus, substantia nigra, locus coeruleus and raphe nuclei. The present study suggests that fibroblast growth factor receptor-1 is expressed preferentially in certain neuronal systems that appear to be under the influence of fibroblast growth factors in the normal brain. The result should facilitate study of the functional significance of fibroblast growth factors in these brain neurons.

• Slack J
• Role of fibroblast growth factors as inducing agents in early embryonic development.
• Mol Reprod Dev. 1994; 39: 118-24
• Display abstract

• To assess the potential role of a molecule in development we need to know three things: 1) what are the biological activities of the molecule, 2) what is its expression pattern, and 3) what are the consequences of removing it from the embryo? In the case of the FGF family in Xenopus embryos we have quite a lot of information about all three questions. Most members of the family can induce mesoderm from isolated animal caps, thus mimicking the natural "ventral vegetal" inducing signal operative in the blastula. This activity can be exerted on isolated, disaggregated cells and does not involve a change in division rate. When overexpressed from injected mRNA, the activity of FGFs depends largely on whether or not they possess a signal sequence, showing the importance of secretion in the inductive process. In addition to the mesoderm-inducing activity, there are effects of overexpression on whole embryos which lead to a suppression of anterior structures. Three types of FGF have so far been cloned from Xenopus: direct homologs of each of the mammalian types FGF-2 and FGF-3, and eFGF ("embryonic FGF"), which is equidistant in sequence from mammalian FGF-4 and FGF-6. Attempts to find homologs of mammalian FGF-5 and FGF-7 in Xenopus have proved unsuccessful. All three types of Xenopus FGF are expressed in early development. FGF-2 and eFGF are present in the oocyte and fertilized egg, and are thus both available at the time of mesoderm induction.(ABSTRACT TRUNCATED AT 250 WORDS)

• Nilsen-Hamilton M
• Introduction: fibroblast growth factors and their receptors in development and disease.
• Mol Reprod Dev. 1994; 39: 41-2

• Maciag T et al.
• Novel mechanisms of fibroblast growth factor 1 function.
• Recent Prog Horm Res. 1994; 49: 105-23

• DeHamer MK, Guevara JL, Hannon K, Olwin BB, Calof AL
• Genesis of olfactory receptor neurons in vitro: regulation of progenitor cell divisions by fibroblast growth factors.
• Neuron. 1994; 13: 1083-97
• Display abstract

• Olfactory receptor neurons are produced continuously in mammalian olfactory epithelium in vivo, but in explant cultures neurogenesis ceases abruptly. We show that in vitro neurogenesis is prolonged by fibroblast growth factors (FGFs), which act in two ways. FGFs increase the likelihood that immediate neuronal precursors (INPs) divide twice, rather than once, before generating neurons; this action requires exposure of INPs to FGFs by early G1. FGFs also cause a distinct subpopulation of explants to generate large numbers of neurons continually for at least several days. The data suggest that FGFs delay differentiation of a committed neuronal transit amplifying cell (the INP) and support proliferation or survival of a rare cell, possibly a stem cell, that acts as a progenitor to INPs.

• Eckenstein FP, Andersson C, Kuzis K, Woodward WR
• Distribution of acidic and basic fibroblast growth factors in the mature, injured and developing rat nervous system.
• Prog Brain Res. 1994; 103: 55-64

• Shao NS, Wang HX
• [Neurotropic effect of fibroblast growth factor on neurons]
• Sheng Li Ke Xue Jin Zhan. 1994; 25: 17-21

• Cui JQ, Liu C, Wang HX
• [Fibroblast growth factor and cardiovascular diseases]
• Sheng Li Ke Xue Jin Zhan. 1994; 25: 76-9

• Gimenez-Gallego G, Cuevas P
• Fibroblast growth factors, proteins with a broad spectrum of biological activities.
• Neurol Res. 1994; 16: 313-6
• Display abstract

• The first two members of the fibroblast growth factor family of proteins were isolated ten years ago. The family now includes nine members. Six of them are biologically and structurally closely related, and often the denomination fibroblast growth factor family refers exclusively to them. It has become progressively evident that the six most similar members of the family induce mitogenesis in practically all mesoderm and neuroectoderm derived cells. In healthy animals only two of these six polypeptides have been detected, which are currently known as acidic and basic fibroblast growth factors, respectively. These two polypeptides are widely distributed in the organism. Evidence has been progressively accumulating that acidic and basic fibroblast growth factors show too, a wide assortment of hormone-like activities.

• Bartlett PF, Dutton R, Likiardopoulos V, Brooker G
• Regulation of neurogenesis in the embryonic and adult brain by fibroblast growth factors.
• Alcohol Alcohol Suppl. 1994; 2: 387-94
• Display abstract

• The development of the nervous system is partly regulated by growth factors, especially by members of the fibroblast growth factor family (FGF) family. Both the proliferation and differentiation of neuronal precursor cells within the developing neural tube appear to depend upon appropriate stimulation with FGF-1 or FGF-2. Recently, it has been shown that cells residing in the sub-ventricular zone of adult brains have characteristics of neuronal precursors: they can respond to FGF-2 by proliferation and FGF-1 induces neuronal differentiation. Thus, it appears that the adult central nervous system has the potential to replace damaged neurons if the precursor cell can be stimulated with the appropriate growth factor. Exposure of the embryo to alcohol appears to diminish the proliferation of the ventricular layer and subsequently reduced the number of neurons generated. This suggests that alcohol may inhibit FGF action, and exogenous administration of factors may provide a mechanism for stimulating or overcoming deficits resulting from such exposure.

• Vainikka S, Joukov V, Wennstrom S, Bergman M, Pelicci PG, Alitalo K
• Signal transduction by fibroblast growth factor receptor-4 (FGFR-4). Comparison with FGFR-1.
• J Biol Chem. 1994; 269: 18320-6
• Display abstract

• We have studied the signal transduction pathways of fibroblast growth factor receptor-4 (FGFR-4) and FGFR-1, which showed virtually identical acidic fibroblast growth factor binding profiles as well as tyrosine autophosphorylation upon activation in transfected L6 rat myoblasts and NIH3T3 mouse fibroblasts. A prominently tyrosyl-phosphorylated doublet of polypeptides of 85 kDa coprecipitated with activated FGFR-4 from both cell lines studied, but these polypeptides were not detected upon immunoprecipitation of activated FGFR-1. Furthermore, FGFR-4 induced only a weak tyrosyl phosphorylation of phospholipase C-gamma and no detectable tyrosyl phosphorylation of the SHC adaptor proteins in contrast to FGFR-1. No phosphorylation of Ras GTPase-activating protein, p64 Syp/PTP1D tyrosine phosphatase, or association of the GRB2 adaptor protein SH2 domain with these receptors was detected. Unlike FGFR-1, FGFR-4 induced only a barely detectable phosphorylation of the cellular serine/threonine kinase Raf-1 and a weaker tyrosyl phosphorylation of mitogen-activated protein kinases than FGFR-1. Despite these differences, stimulation of both receptors resulted in increased DNA synthesis.

• Norioka K, Mitaka T, Mochizuki Y, Hara M, Kawagoe M, Nakamura H
• Interaction of interleukin-1 and interferon-gamma on fibroblast growth factor-induced angiogenesis.
• Jpn J Cancer Res. 1994; 85: 522-9
• Display abstract

• The interaction of interleukin-1 (IL-1) and interferon-gamma (IFN-gamma) actions on several aspects of angiogenesis in vitro and in vivo was studied. The proliferation and migration of human umbilical vein endothelial cells cultured with basic fibroblast growth factor (bFGF) were synergistically inhibited by cotreatment with IL-1 and IFN-gamma. Endothelial cell adhesion to collagen was suppressed by IL-1 and the effect was slightly enhanced by the combination of IL-1 and IFN-gamma. Local administration of IL-1 (10,000 U) and IFN-gamma (1,000 U) inhibited bFGF-induced angiogenesis in the skin of mice, and synergistic inhibitory activity of the combination was demonstrated. Expression of FGF receptors was strongly downregulated by the combination, whereas expressions of epidermal growth factor (EGF) receptors, integrin beta 1 and integrin beta 3 were not. EGF partially abrogated the growth-inhibitory effects of IL-1 and IFN-gamma. These findings indicate that IL-1 and IFN-gamma are each able to act an angiogenesis inhibitor in a situation where FGF plays a major role in angiogenesis, and the activity is synergistically enhanced when they are used in combination.

• Hatai M, Hashi H, Mogi A, Soga H, Yokota J, Yaoi Y
• Stimulation of tyrosine- and serine-phosphorylation of focal adhesion kinase in mouse 3T3 cells by fibronectin and fibroblast growth factor.
• FEBS Lett. 1994; 350: 113-6
• Display abstract

• Phosphorylation of both tyrosine and serine residues of focal adhesion kinase (FAK) was stimulated by the adhesion of BALB/c mouse 3T3 cells to fibronectin, but phosphorylation of threonine was not detectable. Acidic and basic fibroblast growth factors also stimulated the phosphorylation of serine and tyrosine of FAK in cells adhered to poly-L-lysine, but epidermal growth factor and platelet-derived growth factor did not. A fusion protein of fibronectin and basic fibroblast growth factor effectively induced the phosphorylation of FAK. Phosphorylation of FAK in the rat myoblast L-6 cell line, which lacks fibroblast growth factor receptors, was not stimulated by fibroblast growth factors, suggesting that the interaction of fibroblast growth factors with their receptors might cause the phosphorylation of FAK.

• Jin ML, Tong TJ
• [Progress in the study of fibroblast growth factor]
• Sheng Li Ke Xue Jin Zhan. 1994; 25: 157-60

• Swain JL
• Fibroblast growth factor signalling and myogenic development.
• Trans Am Clin Climatol Assoc. 1994; 106: 109-12

• Bellot F
• [A family of receptors with tyrosine kinase activity for growth factors of fibroblasts]
• Pathol Biol (Paris). 1994; 42: 561-5

• Torriglia A, Blanquet PR
• Immunochemical evidence for a fibroblast growth factor receptor in adult retinal optic fiber and synaptic layers.
• Neuroscience. 1994; 60: 969-81
• Display abstract

• Evidence for fibroblast growth factor receptors in the central nervous system has only been obtained using autoradiographic localization of fibroblast growth factor binding sites and messenger RNA. To clarify those neuronal functions that are regulated by fibroblast growth factor receptors, we have localized immunocytochemically the fibroblast growth factor receptor protein in bovine retina, a neural tissue of well-defined structure and function. The extracellular domain of the gene product referred to as fibroblast growth factor receptor 2 was expressed genetically in bacteria to obtain a polyclonal antibody. Positive staining was confined almost exclusively to the synaptic and optic fiber layers. Such a specific association suggests a role for this receptor in modulation of synaptic terminals and ganglion cell axons of the optic nerve, especially with respect to glutamate release.

• Suzui H et al.
• Immunohistochemical reactions for fibroblast growth factor receptor in arteries of patients with moyamoya disease.
• Neurosurgery. 1994; 35: 20-4
• Display abstract

• The cause of moyamoya disease remains unknown, and pathophysiological mechanisms remain uncertain. Basic fibroblast growth factor (FGF) is a pluripotent polypeptide that has been shown to play roles in angiogenesis, tumorigenesis and many other processes. In a previous study, we demonstrated immunohistochemically that the amount of basic FGF was increased above normal in the superficial temporal artery (STA) of patients with moyamoya disease. To clarify the function of basic FGF in moyamoya disease, we have performed an immunohistochemical study of the STA using a polyclonal antihuman FGF receptor antibody and also have tested immunohistochemical reactions for basic FGF. Twelve surgical specimens of the STA from patients with moyamoya disease were studied. Twelve specimens of the STA from skin flaps of patients with other neurological diseases were also investigated for comparison. The sections of the STA from patients with moyamoya disease showed dense and strong FGF receptor and basic FGF immunoreactivity in endothelial cells, in cells scattered in the thickened intima, and in smooth muscle cells in the media. In contrast, the sections of the STA of control patients showed faint basic FGF immunoreactivity. The statistical analysis revealed a significant difference of basic FGF immunoreactivity between moyamoya disease and other neurological diseases (chi 2 = 23; P = 0.0001). Moderately intense FGF receptor immunoreactivity was observed in most control patients. However, the statistical analysis revealed a significant difference of FGF receptor immunoreactivity between moyamoya disease and other neurological diseases (chi 2 = 13.382; P = 0.0012).(ABSTRACT TRUNCATED AT 250 WORDS)

• Sun SS, McFarland DC
• Interaction of fibroblast growth factor with turkey embryonic myoblasts and myogenic satellite cells.
• Comp Biochem Physiol Comp Physiol. 1993; 105: 85-9
• Display abstract

• 1. The interaction of fibroblast growth factor (FGF) with receptors on clonal-derived turkey embryonic and posthatch muscle cells was compared using saturation isotherms. 2. At least two binding sites, including a high affinity receptor and sites of low affinity, which are likely heparin sulfate proteoglycans, were observed on both embryonic myoblasts (EM) and myogenic satellite cells (SC). 3. The FGF binding affinities (Kds) were similar between SC and EM. Receptor Kds were also similar between SC derived from turkeys both selected and unselected for rapid, growth and skeletal muscle accretion rates.

• Hughes SE, Hall PA
• Overview of the fibroblast growth factor and receptor families: complexity, functional diversity, and implications for future cardiovascular research.
• Cardiovasc Res. 1993; 27: 1199-203

• Clements DA, Wang JK, Dionne CA, Goldfarb M
• Activation of fibroblast growth factor (FGF) receptors by recombinant human FGF-5.
• Oncogene. 1993; 8: 1311-6
• Display abstract

• We have purified biologically active recombinant human fibroblast growth factor 5 (FGF-5) from Escherichia coli. In the presence of heparin, recombinant FGF-5 is as active as native growth factor, demonstrating that glycosylation does not significantly potentiate FGF-5 activity. FGF-5 can bind and induce autophosphorylation of human FGF receptors (FGFR) 1 and 2. Competition binding studies show that the KD for FGF-5-FGFR-1 and FGF-5-FGFR-2 interactions are both between 0.5 and 1.5 x 10(-9) M.

• Green JB
• Mesodermal growth factor candidates elected!
• Bioessays. 1993; 15: 129-30

• Hughes SE, Crossman D, Hall PA
• Expression of basic and acidic fibroblast growth factors and their receptor in normal and atherosclerotic human arteries.
• Cardiovasc Res. 1993; 27: 1214-9
• Display abstract

• OBJECTIVE: Acidic and basic fibroblast growth factors (FGF) are potent mitogens for vascular endothelial and smooth muscle cells and also promote angiogenesis. Given that atheroma is associated with the proliferation of a range of cell types, including smooth muscle cells, it has been proposed that these growth factors may play a part in the genesis and/or maintenance of atheromatous lesions. The aim of our study was to examine the distribution of acidic and basic FGF and their known receptor, fibroblast growth factor receptor 1 (FGFR-1), in normal and atherosclerotic arteries. METHODS: Formalin fixed was embedded archival material from a wide range of vessels was examined using a three stage avidin-biotin immunoperoxidase technique and specific polyclonal antibodies to acidic and basic FGF and FGFR-1. RESULTS: In normal arteries basic FGF immunoreactivity was found in medial smooth muscle cells and adventitial blood vessels, the luminal endothelium being non-reactive. Acidic FGF expression was observed predominantly in adventitial fibroblasts, while FGFR-1 expression was confined to the adventitial microvasculature. In early, simple, and advanced lesions acidic and basic FGF were expressed in macrophages and smooth muscle cells, the principal cell types involved in atherosclerotic lesion formation. Increased expression of basic FGF and FGFR-1 was particularly associated with neovascularisation of the atheromatous lesions. CONCLUSIONS: Acidic and basic FGF are expressed in early, simple and advanced atherosclerotic plaques. This suggests that in vivo these growth factors may have a role in the genesis of this disease. Basic FGF and FGFR-1 may potentially be involved in plaque neovascularisation. Such FGF driven angiogenic events may be central to the life threatening complications of atheroma and provide new options for therapeutic intervention.

• Engelmann GL, Dionne CA, Jaye MC
• Acidic fibroblast growth factor and heart development. Role in myocyte proliferation and capillary angiogenesis.
• Circ Res. 1993; 72: 7-19
• Display abstract

• Proliferative growth of the ventricular myocyte (cardiomyocyte) is primarily limited to fetal and early neonatal periods of development. In concert with the neonatal "transition" from proliferative to hypertrophic growth, ventricular remodeling of the nonmyocyte compartment is characterized by increased extracellular matrix synthesis/deposition and capillary angiogenesis. A role for locally generated and bioactive ventricular acidic fibroblast growth factor (aFGF) in these processes is proposed and substantiated by the following: 1) colocalization of aFGF peptide and fibroblast growth factor receptor (flg) transcripts to the developing fetal cardiomyocyte by immunohistochemistry, immunoelectron microscopy, and in situ hybridization, 2) continued localization of aFGF peptide and transcripts to the neonatal/mature cardiomyocyte, and 3) localization of flg immunoreactivity and transcripts to specific neonatal ventricular nonmuscle cell types. Specific ventricular cell types at distinct developmental stages appear to be responsive to ventricular myocyte-derived aFGF (myocytes in the fetal heart and nonmyocytes/endothelial cells in the neonatal heart). These data indicate that expression of aFGF and one of its receptors (flg) are most pronounced in the fetal to early neonatal ventricle, the presence of both suggesting an autocrine/paracrine growth regulatory function. As the animal matures, ventricular capillary angiogenesis may be facilitated by "release" of cardiomyocyte-derived fibroblast growth factors into the surrounding extracellular space/matrix functioning as a "paracrine" angiogenic stimuli. Therefore, the results of our study suggest that myocyte-derived aFGF may function to increase the fetal ventricular cardiomyocyte population in absolute number as well as to facilitate the subsequent increase in capillary angiogenesis that occurs during cardiomyocyte maturation and ventricular remodeling.

• Yeoman LC
• An autocrine model for cell- and matrix-associated fibroblast growth factor.
• Oncol Res. 1993; 5: 489-99
• Display abstract

• While the prototypic fibroblast growth factors (FGFs) are known to have important roles in the stimulation of cell proliferation and angiogenesis, their lack of a conventional secretory signal peptide sequence has presented a challenge to our understanding of the mechanisms by which they are released from cells and subsequently engaged at cell-membrane-localized receptors. The autocrine and paracrine function of polypeptide growth factors has frequently involved the delivery of a large, transmembrane precursor molecule to the cell surface and its subsequent proteolytic release. Through the use of chimeric growth factors, and enhanced proteolytic release of cell-associated populations of growth factor precursors, it has been established that cell-associated growth factor populations can exhibit autocrine and paracrine activity. This commentary will draw upon the potential role of cell- and matrix-associated FGF populations to develop a model for the release of prototypic FGFs from low-affinity sites on the cell surface, or from extracellular matrix, to high-affinity, cellular receptors. It is proposed that cell- and matrix-associated FGF may exhibit autocrine function and provide the capacity to be regulated in a variety of normal and transformed cell systems. It is also proposed that growth factor delivery mechanisms involving cell- and matrix-associated ligand populations represent a specialized form of regulated delivery. This specialized delivery mechanism may confer autocrine advantages upon a variety of growth factor family gene products.

• Turnbull JE, Gallagher JT
• Heparan sulphate: functional role as a modulator of fibroblast growth factor activity.
• Biochem Soc Trans. 1993; 21: 477-82

• Swain S, Wellstein A, Parker B, Lippman M, Steakley C, DeLap R
• Heparin-binding growth factor blockade with pentosan polysulfate.
• Ann N Y Acad Sci. 1993; 698: 63-70

• Brogi E, Winkles JA, Underwood R, Clinton SK, Alberts GF, Libby P
• Distinct patterns of expression of fibroblast growth factors and their receptors in human atheroma and nonatherosclerotic arteries. Association of acidic FGF with plaque microvessels and macrophages.
• J Clin Invest. 1993; 92: 2408-18
• Display abstract

• Because fibroblast growth factors (FGFs) modulate important functions of endothelial cells (EC) and smooth muscle cells (SMC), we studied FGF expression in human vascular cells and control or atherosclerotic arteries. All cells and arteries contained acidic (a) FGF and basic (b) FGF mRNA. Northern analysis detected aFGF mRNA only in one of five control arteries but in all five atheroma tested, while levels of bFGF mRNA did not differ among control (n = 3) vs. plaque specimens (n = 6). Immunolocalization revealed abundant bFGF protein in control vessels (n = 10), but little in plaques (n = 14). In contrast, atheroma (n = 14), but not control arteries (n = 10), consistently exhibited immunoreactive aFGF, notably in neovascularized and macrophage-rich regions of plaque. Because macrophages colocalized with aFGF, we tested human monocytoid THP-1 cells and demonstrated accumulation of aFGF mRNA during PMA-induced differentiation. We also examined the expression of mRNA encoding FGF receptors (FGFRs). All cells and arteries contained FGFR-1 mRNA. Only SMC and control vessels had FGFR-2 mRNA, while EC and some arteries contained FGFR-4 mRNA. The relative lack of bFGF in plaques vs. normal arteries suggests that this growth factor may not contribute to cell proliferation in advanced atherosclerosis. However, aFGF produced by plaque macrophages may stimulate the growth of microvessels during human atherogenesis.

• Aaronson SA
• Growing families of fibroblast growth factors and receptors.
• Jpn J Cancer Res. 1993; 84: 0-0

• Hughes SE, Hall PA
• Immunolocalization of fibroblast growth factor receptor 1 and its ligands in human tissues.
• Lab Invest. 1993; 69: 173-82
• Display abstract

• BACKGROUND: The fibroblast growth factors (FGFs) and their receptors have a wide range of biologic actions in a variety of systems. The best characterized are the prototypical acidic and basic FGFs, both of which have the property of binding to the flg gene product, fibroblast growth factor receptor-1/FGFR-1. Although much is known at the molecular level, the available information about the in vivo distribution of FGFs and their receptors in human tissues is limited. Detailed characterization of the spatial distribution of ligands and receptors is a necessary prerequisite for understanding their function. EXPERIMENTAL DESIGN: Using immunohistochemical techniques, we have defined the tissue specific cellular location of both FGFR-1 and basic and acidic FGFs in serial sections of a broad range of normal human adult tissues. Immunodetection was performed using highly specific polyclonal antibodies to FGFR-1, acidic FGF, and basic FGF and the avidin-biotin immunoperoxidase method. RESULTS: Immunoreactive acidic and basic FGFs were present in almost all tissues examined. A distinct spatial pattern of distribution of these ligands was observed in the kidney, skin, liver, ureter, and the vasculature. FGFR-1 expression tended to be confined to the tissue microvasculature. Interestingly, marked numbers of FGFR-1 reactive vessels were observed in the stomach, salivary glands, lung, and appendix. Intense FGFR-1 positivity was observed in the basal epithelial cell layer of the cervix, palatine tonsil, and respiratory tract as well as breast myoepithelial cells, and the pitutary gland. CONCLUSIONS: We have been able to define the distribution of FGFR-1 and its ligands in normal human tissues. The results will be useful for determining the function of these growth factors and their receptors under both normal physiologic and pathologic conditions.

• Hughes SE, Hall PA
• The fibroblast growth factor and receptor multigene families.
• J Pathol. 1993; 170: 219-21

• Johnson DE, Williams LT
• Structural and functional diversity in the FGF receptor multigene family.
• Adv Cancer Res. 1993; 60: 1-41

• Baird A
• Fibroblast growth factors: what's in a name?
• Endocrinology. 1993; 132: 487-8

• Mascarelli F, Courtois Y
• [Heterocomplex formation between high and low affinity FGF receptors is mediated by the formation of a FGF dimer]
• Bull Cancer. 1993; 80: 786-98
• Display abstract

• Interactions between the two classes of fibroblast growth factor receptors 1) the high affinity receptors (HAR) a membrane glycoprotein containing an intrinsic tyrosine kinase activity, 2) low affinity receptors (LAR) cell surface proteoglycans containing heparan sulfate side chains (HSPG), and aFGF (MW: 15.5 kDa) were studied in bovine lens epithelial (BEL) cells. By Scatchard analysis of the aFGF binding to the BEL cell surface, we show that heparin at 10 micrograms/ml abolishes completely aFGF binding to LAR and reduces by half the number of aFGF HAR. By using cross-linking experiments, aFGF-HAR complexes are present in two forms (150 kDa and 135 kDa). Addition of heparin at 10 micrograms/ml abolishes the formation of the 150 kDa complex and does not affect the 135 kDa complex. Furthermore, binding of aFGF to LAR induces the spontaneous formation of a 31 kDa aFGF dimer. The dimerization process of aFGF on LAR is abolished by addition of heparin. During aFGF internalization at 37 degrees C, we have shown that aFGF-dimer is internalized, accumulated and degraded in the cell as is the 15.5 kDa native form. Heparin at 10 micrograms/ml suppresses specifically aFGF dimer internalization and reduces by half the total amount of internalized aFGF native form. Moreover, after aFGF binding and internalization, the affinity of HAR for aFGF increases concomitantly with its downregulation. Heparin does not seem to affect this phenomenon. All these results strongly suggest that an heteroreceptor dimer-aFGF complex (150 kDa) is formed by one molecule of HAR associated to one molecule of LAR through their respective interaction with a very stable homodimer of aFGF. Such a three component receptor complex induced by FGF dimerization may be a general process of FGF receptor activation which could explain the diversity of the biological response to FGF of different cell type expressing different HAR and LAR or HSPG.

• Itoh N
• [Elucidation of biosynthetic process and regulation mechanism of bioactive peptides and their receptors]
• Yakugaku Zasshi. 1992; 112: 419-31
• Display abstract

• To elucidate the mechanism of proinsulin induction by glucose, we determined the amount of proinsulin messenger ribonucleic acid (mRNA), the subcellular distribution of proinsulin mRNA and the transcriptional activity of proinsulin gene in isolated pancreatic islets of rat. From these results, we concluded that the stimulation of proinsulin synthesis was mainly regulated at the translational level. To elucidate the primary structure of vasoactive intestinal peptide (VIP) precursor, we determined the nucleotide sequence of VIP complementary deoxyribonucleic acid (cDNA). The entire amino acid sequence of the VIP precursor indicates that the precursor protein contains not only VIP but also PHI-like peptide. We also found that the induction of pro-VIP synthesis was achieved by enhancing the transcription rate of VIP gene. We also isolated fibroblast growth factor (FGF) receptor cDNA. The nucleotide sequence of FGF receptor cDNA indicates that the receptor is a transmembrane protein that contains extracellular immunoglobulin-like domains and an intracellular tyrosine kinase domain. We also found that the receptor had two isoforms of the extracellular domains and that the expression of the isoforms was regulated tissue-specifically.

• Prakash S, Wyler DJ
• Fibroblast stimulation in schistosomiasis. XII. Identification of CD4+ lymphocytes within schistosomal egg granulomas as a source of an apparently novel fibroblast growth factor (FsF-1).
• J Immunol. 1992; 148: 3583-7
• Display abstract

• Granulomas that form around Schistosoma mansoni eggs deposited in the liver secrete a variety of fibrogenic factors that may provide a molecular link between chronic inflammation and hepatic fibrogenesis in schistosomiasis. We recently isolated from conditioned medium of egg granuloma cultures a approximately equal to 60-kDa heparin-binding growth factor for fibroblasts. Because this protein is distinct from other defined heparin-binding growth factors, we designated it "fibroblast stimulating factor-1" (FsF-1). We now report that FsF-1 is a lymphokine. We prepared IgG antibody against purified FsF-1 and determined that it did not cross-react with a variety of growth factors or recombinant interleukins. Using two-color flow cytometry of dissociated granuloma cell suspensions, we observed that approximately 20% to 25% of granuloma CD4+ lymphocytes express surface FsF-1. We isolated CD4+ granuloma lymphocytes by FACS and observed that these cells spontaneously secrete into culture supernatant a fibroblast mitogen that is neutralized by anti-FsF-1 antibody. Furthermore, anti-FsF-1 can specifically immunoprecipitate a metabolically labeled protein produced by the granuloma CD4+ lymphocytes. The labeled protein has the same apparent molecular mass (approximately equal to 60 kDa) as FsF-1 purified from granuloma culture supernatants. These findings define CD4+ lymphocytes as a source of FsF-1. Because FsF-1 has biologic and chemical features distinct from most other defined lymphokines and from other heparin-binding growth factors, FsF-1 appears to be a novel lymphokine.

• Givol D, Yayon A
• Complexity of FGF receptors: genetic basis for structural diversity and functional specificity.
• FASEB J. 1992; 6: 3362-9
• Display abstract

• Since 1989, the receptors for fibroblast growth factors (FGFs) were cloned and characterized as a subgroup of the family of receptor tyrosine kinases. Four FGF receptor genes were identified, all of which encode membrane-bound glycoproteins containing three immunoglobulin (Ig) -like domains at the extracellular region, where only two of these domains are involved in ligand binding. Three unique features characterize the FGF receptors: 1) overlapping recognition and redundant specificity, where one receptor may bind with a similar affinity several of the seven known FGFs and one FGF may bind similarly to several distinct receptors. 2) The binding of FGFs to their receptors is dependent on the interaction of FGF with cell surface heparan sulfate proteoglycans. 3) A multitude of isoforms of cell-bound or secreted receptors are produced by the same gene. The gene structure of these receptors revealed two major mechanisms that are responsible for the formation of the diverse forms: alternative mRNA splicing, resulting in deletions or alternate exons usage, and internal polyadenylation, resulting in truncated products. These are reminiscent of mechanisms that also operate in the immunoglobulin family to generate diversity and to produce either secreted or cell-bound molecules. Tissue-specific alternative splicing in FGF receptors allows for the generation of two distinct receptors from a single gene because alternative exons determine the sequence of the COOH-terminal half of the third Ig-like domain involved in ligand binding. This represents a novel genetic mechanism to generate receptor diversity and specificity and to increase receptor repertoire.

• Robak T
• [Current views on the pathogenesis and treatment of spontaneous (primary) myelofibrosis]
• Pol Tyg Lek. 1992; 47: 144-7

• Schlessinger J, Mohammadi M, Margolis B, Ullrich A
• Role of SH2-containing proteins in cellular signaling by receptor tyrosine kinases.
• Cold Spring Harb Symp Quant Biol. 1992; 57: 67-74

• Ledoux D, Gannoun-Zaki L, Barritault D
• Interactions of FGFs with target cells.
• Prog Growth Factor Res. 1992; 4: 107-20
• Display abstract

• Growth factors play a key role in cellular communication, a necessary step for the development of pluricellular organisms. The fibroblast growth factors (FGF) are among these polypeptides and have seven known members: FGF 1 to FGF 7 which are also known as acidic FGF, basic FGF, translation products of oncogenes hst, int 2, FGF 5, FGF 6 and FGF 7 or keratinocyte growth factor (KGF) respectively [1]. The best known and the most abundant in normal adult tissues are acidic and basic FGFs, or FGF 1 and 2 respectively, which have been subjected to extensive studies both in vitro and in vivo. These two factors have almost ubiquitous distribution and a wide spectrum of biological activity including action on cellular proliferation and differentiation, as well as neurotrophic and angiogenic properties [1]. These different activities are induced by triggering specific receptors present at the surface of the target cell. Following this interaction, the FGF-receptor complexes are internalized and activate intracellular pathways. An important effort of investigations has been produced to characterize these receptors and intracellular pathways. It is the purpose of this review to present this work which will focus on FGFs 1 and 2. The existence of two classes of interactions has been reported as early as 1987 [52, 53, 54] suggesting the presence of high and low affinity receptors for FGFs.

• Burrus LW, Zuber ME, Lueddecke BA, Olwin BB
• Identification of a cysteine-rich receptor for fibroblast growth factors.
• Mol Cell Biol. 1992; 12: 5600-9
• Display abstract

• The fibroblast growth factor (FGF) family consists of seven members whose activities are thought to be mediated by multiple receptors. Here we describe the cDNA cloning, expression, and characterization of a cysteine-rich FGF receptor (CFR) that is distinct from previously identified FGF receptors. The deduced amino acid sequence for CFR suggests that it is an integral membrane protein containing a large extracellular domain comprising 16 cysteine-rich repeated units and an intracellular domain of 13 amino acids. No reported sequences exhibit significant homologies to either the repeated extracellular motif or to the entire CFR amino acid sequence. Several CFR transcripts are present in embryonic chick tissue, suggesting that CFR undergoes alternate mRNA splicing or that related genes are present. Chinese hamster ovary cells transfected with the CFR cDNA express a 150-kDa polypeptide that binds FGF-1, FGF-2, and FGF-4 but does not bind several non-FGF family members. The high degree of evolutionary conservation among vertebrate CFRs and its ability to bind three different FGFs with high affinity suggest that this unique receptor plays an important role in FGF biology.

• Pugh-Humphreys RG
• Macrophage-neoplastic cell interactions: implications for neoplastic cell growth.
• FEMS Microbiol Immunol. 1992; 5: 289-308
• Display abstract

• Subcutaneous transplantation of EL4 lymphoma cells within C57BL10 mice evoked an oedematous inflammatory response involving increased leukopoiesis within the bone marrow, a blood leukocytosis, an influx of leukocytes into the transplants and surrounding host connective tissues, and extensive remodelling of surrounding host connective tissues involving fibroplasia and angiogenesis. Dexamethasone not only significantly reduced the numbers of circulating blood leukocytes within C57BL10 mice bearing the subcutaneous EL4 lymphoma transplants, but also reduced the oedematous inflammatory response to the transplants. The decreased influx of inflammatory leukocytes into the site of EL4 lymphoma cell transplantation within the dexamethasone-treated mice, was accompanied by reduced growth of the transplants. Although the EL4 lymphoma cells produce factors with Colony Stimulating Factor activity and with chemotactic activity for cells of the monocyte-macrophage lineage, they do not appear to produce fibroblast growth factors directly but can induce (or stimulate) macrophages to generate fibroblast growth factors in vitro. While not directly inhibiting the growth of subcutaneous fibroblasts in vitro, dexamethasone does suppress the production and/or activity of fibroblast growth factors generated through macrophage-EL4 cell interactions in vitro. The inhibitory effects of dexamethasone on macrophage influx, fibroplasia and angiogenesis within the connective tissue surrounding the EL4 lymphoma transplants appear to be casually related events and would account for the inhibitory effect of dexamethasone on the growth of the lymphoma transplants.

• Thornton SC, Robbins JM, Penny R, Breit SN
• Fibroblast growth factors in connective tissue disease associated interstitial lung disease.
• Clin Exp Immunol. 1992; 90: 447-52
• Display abstract

• Fibrosis is a major cause of morbidity and mortality in chronic inflammatory diseases, especially interstitial pulmonary disorders. Fibroproliferation is an important part of this fibrotic response, and is mediated largely through growth factors such as platelet-derived growth factor (PDGF), insulin-like growth factor (IGF) I and tumour necrosis factor-alpha (TNF-alpha). Although there is some evidence implicating these cytokines in fibrotic disorders, strong evidence in vivo is almost nonexistent. In order to ascertain the role that these factors play in inflammatory lung disorders associated with connective tissue diseases, alveolar mononuclear cells have been obtained from subjects by bronchoalveolar lavage and assessed for the spontaneous release of fibroblast growth factors. The study population consisted of subjects with a variety of different connective tissue disorders, both with and without inflammatory pulmonary complications. It was found that lavage cells spontaneously secreted fibroblast growth factor activity over 24 h with maximum activity detected at 6 to 12 h. Growth factor activity could be detected in most subjects with connective tissue disease-associated inflammatory lung disease and some normal subjects, but the amount of growth factor activity was much higher in the former than in the latter. By means of antibody depletion experiments all growth factor activity from lavage cells of normal patients was attributable to TNF-alpha while patients with interstitial lung disease secreted large amounts of PDGF and fibronectin in addition to TNF-alpha. Approximately 40-50% of the total released growth factor activity could be accounted for by PDGF, and 100% by the combination of PDGF, TNF-alpha and fibronectin. While TNF-alpha is released from the bronchoalveolar lavage cells of many subjects, in addition, many patients with interstitial lung disease also release spontaneously, large amounts of fibroblast growth factor activity attributable to PDGF and fibronectin.

• Jaye M, Schlessinger J, Dionne CA
• Fibroblast growth factor receptor tyrosine kinases: molecular analysis and signal transduction.
• Biochim Biophys Acta. 1992; 1135: 185-99

• Korhonen J et al.
• Five FGF receptors with distinct expression patterns.
• EXS. 1992; 61: 91-100

• Gillespie LL, Paterno GD, Mahadevan LC, Slack JM
• Intracellular signalling pathways involved in mesoderm induction by FGF.
• Mech Dev. 1992; 38: 99-107
• Display abstract

• We have examined the possible role of two signal transducing mechanisms, tyrosine phosphorylation and activation of protein kinase C (PKC), during fibroblast growth factor (FGF)-induced mesoderm induction in Xenopus. Tyrosine phosphorylation was examined through the use of a monoclonal anti-phosphotyrosine antibody. This antibody was shown to recognize the FGF receptor crosslinked to radioiodinated FGF. We also studied the response of Xenopus ectodermal explants to sodium orthovanadate, a compound that has been shown to elevate intracellular phosphotyrosine levels. Thirty percent of explants cultured in 100 microM vanadate were induced. In addition, vanadate synergized with FGF to give inductions that were more dorsal in nature than either vanadate or FGF alone. The role of PKC was evaluated by measuring PKC activity during mesoderm induction by FGF and by examining the effect of the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA) on explants. TPA did not induce mesoderm, however, activation of PKC was detected in FGF-treated explants. Therefore, activation of the PKC pathway alone is not sufficient for mesoderm induction. Simultaneous treatment with TPA and FGF resulted in a significant inhibition of mesoderm induction by FGF, suggesting that activation of PKC could be part of a negative feedback mechanism. In contrast, TPA had no effect on induction by activin A.

• Shiokawa K, Asano M, Shiozaki C
• [Function, molecular structure and gene expression of fibroblast growth factor (FGF/HBGF)]
• Nippon Rinsho. 1992; 50: 1893-901
• Display abstract

• The structure and function of members of the fibroblast growth factor (FGF) gene family and their receptors are reviewed. All the member of this gene family bind heparin, and therefore, are also called the as heparin-binding growth factor (HBGF). In this review, the structural features of FGF/HBGF are summarized first, and general features of the structure and function of their receptors are then described briefly. After biological effects of FGF/HBGF on adult-type tissues and cultured cells are reviewed, effects of FGF on the mesodermal induction in amphibian (Xenopus laevis) embryonic system are reviewed in more detail. Emphasis is given on the experiments with Xenopus animal cap assay system and also on the injection into Xenopus fertilized eggs of mRNA of dominant defect mutant of FGF receptor, which leads to the formation of embryos with abnormal axial mesoderm.

• Habazettl J, Gondol D, Wiltscheck R, Otlewski J, Schleicher M, Holak TA
• Structure of hisactophilin is similar to interleukin-1 beta and fibroblast growth factor.
• Nature. 1992; 359: 855-8
• Display abstract

• The fast reaction of the actin-based cytoskeleton in motile cells after stimulation with a chemoattractant requires a signal-transduction chain that creates a very specific environment at distinct regions beneath the plasma membrane. Dictyostelium hisactophilin, a unique actin-binding protein, is a submembranous pH sensor that signals slight changes of the H+ concentration to actin by inducing actin polymerization and binding to microfilaments only at pH values below seven. It has a relative molecular mass of 13.5K and its most unusual feature is the presence of 31 histidine residues among its total of 118 amino acids. The transduction of an external signal from the plasma membrane to the cytoskeleton is poorly understood. Here we report the protein's structure in solution determined by nuclear magnetic resonance spectroscopy. The nuclear Overhauser effect intensities of the three-dimensional nuclear Overhauser spectra were used directly in the calculations. The overall folding of histactophilin is similar to that of interleukin-1 beta and fibroblast growth factor, but the primary amino-acid sequence of hisactophilin is unrelated to these two proteins.

• Partanen J, Vainikka S, Korhonen J, Armstrong E, Alitalo K
• Diverse receptors for fibroblast growth factors.
• Prog Growth Factor Res. 1992; 4: 69-83
• Display abstract

• The development and maintenance of multicellular organisms requires a complex interplay between cells in different tissues. Many of the factors mediating cell-cell communication are polypeptides, which were originally identified because of their ability to stimulate cell growth. In addition to growth signalling several of these factors have been observed to modulate cell survival, chemotaxis and differentiation both in vitro and in vivo. Fibroblast growth factors are a good example of polypeptide mitogens eliciting a wide variety of responses depending on the target cell type. Our knowledge of the cell surface receptors mediating the effects of FGFs has recently expanded remarkably. Perhaps not surprisingly, the complexity of the FGF family and FGF induced responses is reflected as diversity and redundancy of the FGF receptors.

• Avivi A, Skorecki K, Yayon A, Givol D
• Promoter region of the murine fibroblast growth factor receptor 2 (bek/KGFR) gene.
• Oncogene. 1992; 7: 1957-62
• Display abstract

• Fibroblast growth factors (FGFs) and their receptors play an important role in cell growth, angiogenesis and embryonal development. Four distinct genes encoding fibroblast growth factor receptors (FGFRs) were identified: flg, encoding FGFR1, bek encoding FGFR2, and the genes for FGFR3 and FGFR4. Both FGFR2 and keratinocyte growth factor receptor (KGFR) are encoded by the same gene, bek. To study the regulation of expression of the FGF receptors we analysed the DNA sequence flanking the 5' region of the cDNA of murine FGFR2 to seek elements that control its transcription. A 5-kbp fragment containing the 5' end of the cDNA was isolated from mouse genomic library and used to map the promoter region. We found that the sequence encoding the 5' non-translated region of the FGFR2/KGFR cDNA contains an intron located 210 bp upstream from the translation start site. Using RNAase protection and primer extension, we identified the mRNA start 37 bp upstream from the beginning of the bek cDNA. The promoter activity was found to reside in a 1.3-kbp fragment upstream from the cDNA, and deletion mapping further localized the promoter to a 0.7-kbp fragment. The sequence of this region shows high G+C content (62%), which is particularly emphasized in the 200 bp upstream from the mRNA start (80% G+C). This region contains the CCGCCC, GGGCGG AND GGAGG motifs also found in promoters of other growth factor receptors. Neither TATA nor CAAT boxes were found near the RNA start site. The characterization of this promoter will allow studies of the regulation of expression of the FGFR2 during development and in pathophysiological states. The differences between the promoter sequence of the gene for FGFR2 (bek) and FGFR1 (flg) may explain their differential expression during development.

• Denholm EM
• Continuous secretion of monocyte chemotactic factors and fibroblast growth factors by alveolar macrophages following a single exposure to bleomycin in vitro.
• Am J Pathol. 1992; 141: 965-71
• Display abstract

• It has been shown in previous studies that alveolar macrophages incubated with bleomycin in vitro for 2 to 18 hours secrete monocyte chemotactic factors and fibroblast growth factors (MDGF). The purpose of the current experiments was to determine if alveolar macrophages similarly stimulated with bleomycin would continue to secrete these factors once the stimulus was removed. Alveolar macrophages from normal rats were exposed to bleomycin for 18 hours after which bleomycin was removed and macrophages maintained in culture for 35 days. Conditioned medium (CM) was collected and assayed at weekly intervals. In comparison with nonstimulated controls, bleomycin-stimulated macrophages secreted greater amounts of both monocyte chemotactic factors and MDGF for 35 days after exposure to bleomycin; with a significant difference noted between bleomycin and control macrophages for the first 21 days (P less than 0.02). In agreement with past work, the chemotactic activity in bleomycin-CM was due to fibronectin, as evidenced by the almost complete inhibition of activity by anti-fibronectin antibodies. The time course of secretion of chemotactic and growth factors after a single exposure to bleomycin in vitro was similar to that induced by in vivo exposure of macrophages to this drug. The data suggest that a similar direct activation of macrophages by bleomycin may promote the long-term production of these factors in vivo, resulting in continued monocyte recruitment and promotion of fibroblast proliferation in fibrotic lungs.

• Brem H, Klagsbrun M
• The role of fibroblast growth factors and related oncogenes in tumor growth.
• Cancer Treat Res. 1992; 63: 211-31

• Lafage M et al.
• Fusion and amplification of two originally non-syntenic chromosomal regions in a mammary carcinoma cell line.
• Genes Chromosomes Cancer. 1992; 5: 40-9
• Display abstract

• The FLG/FGFRI gene, encoding a receptor for members of the FGF family, is located at 8p11.2-p12. It is amplified, overexpressed, and not grossly rearranged in the MDA-MB-134 breast carcinoma cell line, whereas other genes from the pericentromeric 8p region are not amplified. The FGF4/HSTFI gene, located at 11q13, is also amplified with a substantial portion of the 11q13 region, but is not overexpressed in MDA-MB-134 cells. In this cell line, amplified sequences constitute a large homogeneously staining region (HSR) which is part of a marker chromosome containing chromosome 8 and chromosome 11 sequences. Using probes for the FGF4/HSTFI and the FLG/FGFRI genes in fluorescence chromosomal in situ hybridization, we show that the HSR contains de novo fused and amplified 11q13 and 8p11-p12 sequences associated in a complex structure containing approximately the same number of FGF4 and FGFRI genes. The significance of this genetic abnormality for MDA-MB-134 cells, and for breast carcinogenesis in general, is unknown, but may underlie a particular type of oncogene activation.

• Blanckaert V, Hondermarck H, Boilly-Marer Y
• Differential expression of fibroblast growth factor-like molecules and their receptors: a putative role in tissue regeneration and growth in annelids.
• Monogr Dev Biol. 1992; 23: 28-39

• Hondermarck H, Boilly B
• Characterization of fibroblast growth factor binding in regenerating limb blastemas of axolotls.
• Monogr Dev Biol. 1992; 23: 110-5

• Mitsui Y, Imamura T
• [Growth regulation of vascular cells by FGFs and a novel ECGF]
• Tanpakushitsu Kakusan Koso. 1991; 36: 1237-46

• Hearn MT
• Structure and function of the heparin-binding (fibroblast) growth factor family.
• Baillieres Clin Endocrinol Metab. 1991; 5: 571-93

• Gospodarowicz D
• Biological activities of fibroblast growth factors.
• Ann N Y Acad Sci. 1991; 638: 1-8

• Suzuki F
• [Growth factors of chondrocytes]
• Tanpakushitsu Kakusan Koso. 1991; 36: 1259-67

• Birnbaum D et al.
• Role of FGFs and FGF receptors in human carcinogenesis.
• Ann N Y Acad Sci. 1991; 638: 409-11

• Shing Y
• Biaffinity chromatography of fibroblast growth factors.
• Methods Enzymol. 1991; 198: 91-5

• Terrell GE, Swain JL
• Indirect angiogenic agents do not release fibroblast growth factors from extracellular matrix.
• Matrix. 1991; 11: 108-14
• Display abstract

• Vascular growth factors are categorized as either primary or secondary angiogenic factors. Primary angiogenic agents such as fibroblast growth factors, not only induce the complete angiogenic response, but also stimulate the individual components of vascular growth. Secondary angiogenic agents can induce vascular growth, but they do not act through the direct stimulation of endothelial proliferation, migration, and protease production. Since fibroblast growth factors are known to bind to components of the extracellular matrix, we assessed whether secondary agents act through liberating growth factors from matrix storage sites. The study utilized L6 skeletal myoblasts in culture, which we demonstrated were capable of synthesizing extracellular matrix containing heparin binding endothelial mitogens. The heparin-binding mitogenic activity accumulated in a time-dependent fashion, and matrix extracts contained a protein with immunologic identity to acidic fibroblast growth factor. The ability of secondary angiogenic agents and related compounds including adenosine, inosine, hypoxanthine, nicotinamide, lactic acid, phorbol esters, prostaglandin E2, and copper (at concentrations of 1 microM and 1 mM) to release heparin binding mitogenic activity from the matrix was evaluated. The results demonstrate that although heparin is capable of releasing heparin-binding growth factors from extracellular matrix storage sites in a dose dependent fashion, none of the known secondary angiogenesis factors are capable of functioning in a similar fashion. Thus these secondary angiogenic factors do not appear to exert their effect through increasing the bioavailability of preformed heparin-binding growth factors sequestered in the extracellular matrix. The mechanism(s) whereby these agents induce vascular growth remains to be elucidated.

• Jacquot-Dourges MA, Zhou FL, Muller D, Jozefonvicz J
• Affinity chromatography of fibroblast growth factors on coated silica supports grafted with heparin.
• J Chromatogr. 1991; 539: 417-24
• Display abstract

• Dextran-coated silica beads are excellent supports for affinity chromatography of proteins. They can be easily grafted using conventional coupling methods with different active ligands, such as heparin. Fibroblast growth factors develop specific interactions with heparin through well-defined amino acids sequences. The heparin-dextran coated silica phases exhibit an affinity for these growth factors. Under our experimental conditions, the basic form can be absorbed on the solid support at a moderate salt concentration (0.5 M sodium chloride) and can be selectively desorbed by increasing the ionic strength of the eluent. The purification performances of such phases are compared to those obtained on the heparin grafted soft gels. Because of their mechanical properties, the dextran-coated silica supports were also used in high-performance affinity chromatography to purify fibroblast growth factors from a bovine brain crude extract.

• Nishikawa K, Yoshitake Y, Minemura M, Yamada K
• [Functions of heparin binding growth factor (HBGF or FGF) produced by tumor cells]
• Tanpakushitsu Kakusan Koso. 1991; 36: 1364-70

• Robinson CJ
• Tailoring and targeting fibroblast growth factors.
• Trends Biotechnol. 1991; 9: 147-8

• Giacobini MM, Hoffer B, Olson L
• Fibroblast growth factors influence central nervous system development. Evidence from intraocular grafts.
• Ann N Y Acad Sci. 1991; 638: 484-6

• Orr-Urtreger A, Givol D, Yayon A, Yarden Y, Lonai P
• Developmental expression of two murine fibroblast growth factor receptors, flg and bek.
• Development. 1991; 113: 1419-34
• Display abstract

• Developmental expression of two closely related fibroblast growth factor receptors, bek and flg, is described from early postimplantation until advanced organogenesis. Transcripts of bek and flg were first seen in the primitive ectoderm of egg-cylinder-stage embryos. Later, starting with somitogenesis, and then throughout embryogenesis, they were actively transcribed both in the mesoderm and neuroectoderm. Bek was expressed also in the surface ectoderm and in various epithelia, whereas flg expression was restricted mainly to the mesenchyme. In the limb bud bek transcripts displayed a gradient-like distribution and appeared earlier than flg. The two receptors, in contrast to their almost identical ligand binding specificity, displayed distinct spatial specificities throughout development, suggesting that developmental localization may contribute to functional specificity. The role of bek and flg in gastrulation and in epithelial-mesenchymal interactions of organogenesis will be discussed.

• Hendry IA, Belford DA
• Lack of retrograde axonal transport of the heparin-binding growth factors by chick ciliary neurones.
• Int J Dev Neurosci. 1991; 9: 243-50
• Display abstract

• In view of the likelihood that the heparin-binding growth factors of fibroblast growth factors are neurotrophic for the neurones of the chick ciliary ganglion a study has been made of the retrograde axonal transport of the acidic and basic fibroblast growth factors by these neurones. No high capacity retrograde axonal transport of these molecules was seen. The amount of transport was equivalent to the low level seen with many proteins such as horse radish peroxidase or bovine serum albumin rather than the convincing transport seen for nerve growth factor in the sympathetic system. The iodinated proteins retained their ability to bind to neuronal receptors. Thus, if the fibroblast growth factors are neurotrophic in the ciliary ganglion, they may exert their action by mechanisms other than the retrograde axonal transport of the factor itself.

• Cuevas P, Carceller F, Ortega S, Zazo M, Nieto I, Gimenez-Gallego G
• Hypotensive activity of fibroblast growth factor.
• Science. 1991; 254: 1208-10
• Display abstract

• Acidic and basic fibroblast growth factors (FGFs) are members of a family of proteins that are broad-spectrum mitogens, have diverse hormone-like activities, and function in tumorigenesis. FGF's ability to raise the concentration of intracellular calcium ion suggests that FGF could induce the synthesis of endothelium-derived relaxing factor (EDRF) and consequently vasodilation. Systemic administration of FGF decreased arterial blood pressure. This effect was mediated by EDRF and by adenosine triphosphate-sensitive potassium ion channels. The hypotensive effect of FGF was segregated from its mitogenic activity by protein engineering. These results extend the range of FGF autocrine activities and potential therapeutic applications, emphasize the role of endothelium as an arterial blood pressure--regulating organ, and provide insight on the structural basis of FGF functions.

• Schelling ME
• FGF mediation of coronary angiogenesis.
• Ann N Y Acad Sci. 1991; 638: 467-9

• Ledoux D, Mereau A, Jaye M, Miskulin M, Barritault D, Courty J
• Adult brain but not kidney, liver, lung, intestine, and stomach membrane preparations contain detectable amounts of high-affinity receptors to acidic and basic growth factors.
• Ann N Y Acad Sci. 1991; 638: 397-9

• Halaban R, Funasaka Y, Lee P, Rubin J, Ron D, Birnbaum D
• Fibroblast growth factors in normal and malignant melanocytes.
• Ann N Y Acad Sci. 1991; 638: 232-43

• Shieh MT, Spear PG
• Fibroblast growth factor receptor: does it have a role in the binding of herpes simplex virus?
• Science. 1991; 253: 208-10

• Clement-Lacroix P, Friteau L, Damais C
• Activation of human dermal fibroblasts by fibroblast growth factors (FGFs) and/or interleukin-1 beta (IL-1 beta).
• Lymphokine Cytokine Res. 1991; 10: 111-4
• Display abstract

• Human dermal fibroblast proliferation was dependent of growth factors such as interleukin-1 (IL-1) and fibroblast growth factors (FGFs). These mediators, which induce proliferation of the cells in culture, were able to synergize when added in combination. This synergistic effect seems to be restricted to the mitogenic activity since IL-1-induced PGE2 release and interferon-beta 1 (IFN-beta 1) production by fibroblasts was inhibited in the presence of FGFs, which by themselves were unable to stimulate the production of IFN-beta 1 and the release of arachidonate metabolites from the fibroblasts even at high concentration.

• Ueno N, Nishimatsu S, Murakami K, Asashima M
• [Growth factors in early development]
• Tanpakushitsu Kakusan Koso. 1991; 36: 1292-9

• Wang HX
• [Heparin-binding growth factors]
• Sheng Li Ke Xue Jin Zhan. 1991; 22: 256-8

• Doctor JS, Hoffmann FM, Olwin BB
• Identification of a fibroblast growth factor-binding protein in Drosophila melanogaster.
• Mol Cell Biol. 1991; 11: 2319-23
• Display abstract

• As assessed by competitive binding and protein-crosslinking experiments, Drosophila melanogaster cells possess basic fibroblast growth factor (bFGF)-specific binding proteins that are similar to FGF receptors on vertebrate cells in molecular weight and binding affinity; these D. melanogaster cells, however, have no detectable binding proteins for acidic fibroblast growth factor (aFGF). Consistent with the presence of bFGF-specific binding proteins, D. melanogaster cells degrade bFGF but not aFGF. These results indicate the conservation of heparin-binding growth factors and receptors between vertebrates and D. melanogaster.

• van Zoelen EJ
• Role of heparin-binding growth factors in embryonic tumors.
• Recent Results Cancer Res. 1991; 123: 155-63

• Wagner JA
• The fibroblast growth factors: an emerging family of neural growth factors.
• Curr Top Microbiol Immunol. 1991; 165: 95-118

• Courtin F, Liva P, Gavaret JM, Toru-Delbauffe D, Pierre M
• Induction of 5-deiodinase activity in astroglial cells by 12-O-tetradecanoylphorbol 13-acetate and fibroblast growth factors.
• J Neurochem. 1991; 56: 1107-13
• Display abstract

• In the brain, 5'-deiodinase (5'-D) is responsible for the metabolic activation of thyroxine (T4) into 3,5,3'-triiodothyronine (T3) and 5-deiodinase (5-D) deiodinates T4 and T3 into inactive metabolites. This study examines the effects of factors known to induce astroglial 5'-D activity on the 5-D activity in cultured rat astroglial cells. The potencies of these factors were compared after 8 h of incubation, when stimulations by these factors near their maximal effects. 12-O-Tetradecanoylphorbol 13-acetate (TPA) at 10(-7) M was a potent inducer of 5-D activity, producing a 30- to 80-fold increase after 8 h. The maximal effect of TPA was observed after about 14 h. The TPA stimulation of 5-D activity was not dependent on glucocorticoids, unlike 5'-D activity. In comparison with TPA, 8-bromo-cyclic AMP (10(-3) M) was a poor inducer of 5-D activity whereas it is an excellent inducer of 5'-D activity. It produced a 2- to 20-fold increase in 5-D activity after 8 h. Natural acidic fibroblast growth factor (20 ng/ml) produced a degree of stimulation similar to that of TPA after 8 h. The maximal effect of acidic fibroblast growth factor was observed after about 16 h (until a 120-fold increase). Recombinant acidic fibroblast growth factor also induced 5-D activity. Basic fibroblast growth factor was less potent than acidic fibroblast growth factor for increasing 5-D activity (maximal increase by 40- to 50-fold after 8 h).(ABSTRACT TRUNCATED AT 250 WORDS)

• Robinson CJ
• Multiple receptors found for the growing FGF family.
• Trends Pharmacol Sci. 1991; 12: 123-4

• Logan A
• CNS growth factors.
• Br J Hosp Med. 1990; 43: 428-37
• Display abstract

• The fundamental importance of polypeptide growth factors in regulating growth and maintaining viability within the CNS is now widely acknowledged. Although the physiological roles of individual growth factors remain to be defined, increasing evidence suggests involvement within the CNS in development, maintenance of neurones, injury responses, the pathogenesis of degenerative diseases and possibly oncogenesis. This review focuses on nerve growth factor and the fibroblast growth factors.

• Slack JM, Darlington BG, Gillespie LL, Godsave SF, Isaacs HV, Paterno GD
• Mesoderm induction by fibroblast growth factor in early Xenopus development.
• Philos Trans R Soc Lond B Biol Sci. 1990; 327: 75-84
• Display abstract

• In early amphibian development the mesoderm is formed around the equator of the blastula in response to inductive signals from the endoderm. At the time of its formation the mesoderm consists of a large 'ventral type' zone and a small 'organizer' zone. A screen of candidate substances showed that a small group of heparin binding growth factors (HBGFs) were active as mesoderm inducing agents in vitro. The fibroblast growth factors (aFGF and bFGF) and embryonal carcinoma derived growth factor (ECDGF) all show similar potency and can produce ventral inductions at concentrations above about 100 pm. Single blastula ectoderm cells can be induced and will differentiate in a defined medium to form mesodermal tissues and all inner blastula cells are competent to respond to the factors. Inducing activity can be extracted from Xenopus blastulae and can be purified by heparin affinity chromatography. Antibody neutralization and Western blotting experiments identify this activity as bFGF. The amounts present are small but would be sufficient to evoke ventral inductions in vivo. It is not yet known whether the bFGF is localized to the endoderm, although it is known that inducing activity secreted by endodermal cells can be neutralized by heparin. The competence of ectoderm to respond to FGF rises from about the 128-cell-stage and falls again by the onset of gastrulation. This change is paralleled by a rise and fall of binding of 125I-labelled aFGF. Chemical cross-linking reveals that this binding is attributable to a receptor of molecular mass about 130 kilodaltons (kDa). The receptor is present both in the marginal zone, which responds to the signal in vivo, and in the animal pole region, which is not induced in vivo but which will respond to HBGFs in vitro. In intact embryos we believe that the ventral type mesoderm forms the somites, kidney and other intermediate structures as well as the blood islands of the ventral midline. These intermediate structures are induced as a function of distance from the organizer in a process called 'dorsalization'. Lithium salts have a dorsalizing effect on whole embryos and also on explants from the ventral marginal zone, causing them to form large blocks of muscle. Lithium will also cause large muscle blocks to form when applied to ectoderm explants together with FGF. It is difficult to extend these results directly to mammalian embryos, but we have shown that the products of the murine int-2 gene and of the human k-fgf genes are active as mesoderm inducing factors.

• Eckenstein FP, Esch F, Holbert T, Blacher RW, Nishi R
• Purification and characterization of a trophic factor for embryonic peripheral neurons: comparison with fibroblast growth factors.
• Neuron. 1990; 4: 623-31
• Display abstract

• The validation of NGF as a physiologically important neurotrophic factor has led to intense efforts to identify novel polypeptide growth factors for neurons. We report here the details of a greater than 80,000-fold purification of a neurotrophic molecule, referred to as growth-promoting activity (GPA), from chicken sciatic nerves. The final product of the purification migrated as a protein band of 21.5 kd, its apparent pI was approximately 4.8, and the ED50 of the most active preparation was approximately 10 pg/ml. Amino acid sequence of a proteolytic digestion fragment of GPA revealed homology with the recently published sequences for rabbit and rat sciatic nerve CNTF. Thus this molecule may be the chicken form of CNTF. Analysis of the specificity of action of GPA showed that, in addition to E8 ciliary ganglion neurons, the factor was able to support short-term survival of E8 dorsal root ganglion and E12 sympathetic neurons. This range of specificities of biological action was also seen with both acidic and basic FGF in the presence of heparin. The biological activity of GPA differed from that of FGF in that it was not potentiated by heparin and did not stimulate mitogenesis in chick fibroblasts.

• Klagsbrun M
• The affinity of fibroblast growth factors (FGFs) for heparin; FGF-heparan sulfate interactions in cells and extracellular matrix.
• Curr Opin Cell Biol. 1990; 2: 857-63

• Goldfarb M
• The fibroblast growth factor family.
• Cell Growth Differ. 1990; 1: 439-45

• Yuen SN, Folkman J, Weisz PB, Joullie MM, Ewing WR
• Affinity of fibroblast growth factors for beta-cyclodextrin tetradecasulfate.
• Anal Biochem. 1990; 185: 108-11
• Display abstract

• beta-Cyclodextrin tetradecasulfate was found to have a very strong affinity for fibroblast growth factor (FGF) and could substitute for heparin in FGF purification. Basic FGF was purified about 200,000-fold from a rat chondrosarcoma using a method of biaffinity chromatography in which the beta-cyclodextrin tetradecasulfate polymer was mixed with copper-Sepharose. This method takes advantage of the strong affinity of FGF for both beta-cyclodextrin tetradecasulfate and copper.

• Isacchi A, Bergonzoni L, Sarmientos P
• Complete sequence of a human receptor for acidic and basic fibroblast growth factors.
• Nucleic Acids Res. 1990; 18: 1906-1906

• Casscells W et al.
• Isolation, characterization, and localization of heparin-binding growth factors in the heart.
• J Clin Invest. 1990; 85: 433-41
• Display abstract

• Acidic and basic fibroblast growth factors (aFGF and bFGF) are angiogenic polypeptide mitogens for cells of mesodermal and neuroectodermal origin. In this report we describe the purification from several normal human hearts (including a very fresh, nonischemic sample) of heparin-binding, acid-, heat- and trypsin-sensitive 14-18-kD peptides that crossreact with antisera against aFGF and bFGF. Further evidence includes (a) prevention of mitogenicity by protamine and by anti-bFGF, (b) displacement of 125I-bFGF from cell membranes, and (c) stimulation of capillary endothelial cell migration. Specific immunohistochemistry localized bFGF to endothelial cells and, surprisingly, to cardiac myocytes, with almost no immunoreactivity in smooth muscle cells. These peptides may function in cardiac embryogenesis, hypertrophy, atherogenesis, angiogenesis, and wound healing, and may also have endocrine, neurotropic, or vasomotor functions.

• Schneider MD, Parker TG
• Cardiac myocytes as targets for the action of peptide growth factors.
• Circulation. 1990; 81: 1443-56

• Dourges MA, Gulino D, Courty J, Badet J, Barritault D, Jozefonvicz J
• Affinity chromatography of fibroblast growth factors on substituted polystyrene.
• J Chromatogr. 1990; 526: 35-45
• Display abstract

• The heparin-binding growth factors aFGF and bFGF (acidic and basic fibroblast growth factor) from crude bovine brain extract were co-eluted with purified [125I]aFGF and/or [125I]bFGF as tracers from heparin-Sepharose and from several insoluble substituted polystyrenes used as stationary phases in low-pressure affinity chromatography. The ability of the resins to isolate FGFs was determined by measuring the eluted radioactivity. It was demonstrated that the various substituted polystyrene resins retain [125I]aFGF and [125I]bFGF with different specificities according to the chemical nature of the substituted groups bound to the polystyrene support. Bifunctional resins substituted with sulphonate and phenylalanine sulphamide groups adsorbed both [125I]aFGF and [125I]bFGF whereas bifunctional resins substituted with sulphonate and sulphamide serine adsorbed only [125I]bFGF. These stationary phases could be adapted to high-performance affinity chromatography and used to isolate growth factors of the FGF family.

• Benharroch D, Birnbaum D
• Biology of the fibroblast growth factor gene family.
• Isr J Med Sci. 1990; 26: 212-9
• Display abstract

• The biology of the fibroblast growth factor (FGF) gene family is reviewed. The structure, expression and known and putative functions of the seven members of this family characterized to date are discussed. These genes include: acidic and basic FGF, the INT.2 and HST proto-oncogenes and the recently characterized FGF.5, FGF.6 and KGF genes. These genes play a role in various processes such as cell proliferation, angiogenesis and embryogenesis, and possibly carcinogenesis.

• Gospodarowicz D
• Fibroblast growth factor and its involvement in developmental processes.
• Curr Top Dev Biol. 1990; 24: 57-93

• Houssaint E et al.
• Related fibroblast growth factor receptor genes exist in the human genome.
• Proc Natl Acad Sci U S A. 1990; 87: 8180-4
• Display abstract

• We have isolated, from a human tumor cDNA library, a gene encoding a putative receptor-like protein-tyrosine kinase that we call TK14. The amino acid sequence of the TK14 protein is closely related to the available partial sequence of the mouse protein bek, and more distantly related to the sequences of a chicken basic fibroblast growth factor receptor (73% sequence homology) and the apparent human equivalent of this receptor, the FLG protein (encoded by the fms-like tyrosine kinase gene). Overexpression of the TK14 protein by transfection of COS-1 cells with the corresponding cDNA in a simian virus 40-based expression vector leads to the appearance of new cell-surface binding sites for both acidic and basic fibroblast growth factors. This has been demonstrated by specific binding assays and chemical cross-linking experiments using 125I-labeled growth factors. It appears, therefore, that the human genome contains at least two distinct genes, for TK14 and FLG, that code for related fibroblast growth factor receptors.

• Zheng WX, Butwell TJ, Heckert L, Griswold MD, Bellve AR
• Pleiotypic actions of the seminiferous growth factor on two testicular cell lines: comparisons with acidic and basic fibroblast growth factors.
• Growth Factors. 1990; 3: 73-82
• Display abstract

• Bovine seminiferous growth factor (SGF) stimulated different pleiotypic responses in TM3 Leydig and TM4 Sertoli cells from those obtained with bovine acidic and basic fibroblast growth factors (aFGF, bFGF). First, the three growth factors had distinct mitogenic effects. SGF increased TM3 and TM4 cell numbers, whereas aFGF was mitogenic only for TM3 cells, and bFGF was inactive on both cell lines. Second, only SGF and bFGF stimulated TM4 cells to produce 3- and 2-fold, respectively, greater levels of extracellular, sulfated glycoprotein-1. By Northern analyses, SGF increased the steady-state levels of sulfated glycoprotein-1 mRNA approximately 1.34-fold relative to those of actin mRNA during a 48-hr period. Third, the cell lines secreted different [35S]methionine-labeled proteins. With TM3 cells, some proteins were secreted specifically, whereas other were up- or down-regulated differentially in response to SGF, aFGF, or bFGF. Likewise, with TM4 cells, the three growth factors induced qualitative and quantitative changes in the secretion of specific proteins. On immunoblots, SGF did not bind antibodies specific for an internal domain of aFGF or FGF-5, nor those directed against N-terminal, internal, and C-terminal regions of bFGF. These data suggest SGF, aFGF, and bFGF acted on TM3 Leydig and TM4 Sertoli cells through different receptors and/or diverging pathways of signal transduction to induce different pleiotypic effects.

• Finklestein SP et al.
• Growth factor expression after stroke.
• Stroke. 1990; 21: 1224-1224
• Display abstract

• Fibroblast growth factors are polypeptides with potent trophic effects on central nervous system cells. Both acidic and basic forms of fibroblast growth factor are found in the mammalian brain. We have examined the expression of these factors after focal brain injury or stroke. After infarction of the lateral cerebral cortex in the mature rat brain, we found a twofold to threefold increase during the first 3 weeks after stroke in levels of fibroblast growth factors in tissue surrounding infarcts. This increase persisted for at least 2 months and appeared mainly to be due to increased levels of basic, but not acidic, fibroblast growth factor. Because of its gliotrophic, angiogenic, and neuronotrophic properties, basic fibroblast growth factor may play an important role in the cascade of cellular reactions that contributes to wound healing and functional recovery after stroke.

• Faktorovich EG, Steinberg RH, Yasumura D, Matthes MT, LaVail MM
• Photoreceptor degeneration in inherited retinal dystrophy delayed by basic fibroblast growth factor.
• Nature. 1990; 347: 83-6
• Display abstract

• Numerous inherited retinal degenerations exist in animals and humans, in which photoreceptors inexplicably degenerate and disappear. In RCS rats with inherited retinal dystrophy, the mutant gene is expressed in the retinal pigment epithelial (RPE) cell, and leads to the loss of photoreceptor cells. Photoreceptors can be rescued from degeneration if they are juxtaposed to wild-type RPE cells in experimental chimaeras or by the transplantation of RPE cells from normal rats. In both cases, the rescue effect extends beyond the immediate boundaries of the normal RPE cells, suggesting trophic action of a diffusible factor(s) from the normal RPE cells. We considered that the fibroblast growth factors, aFGF and bFGF, might have such a trophic role as they are found in the retina and RPE cells; bFGF acts as a neurotrophic agent after axonal injury in several regions of the central nervous system, and bFGF induces retinal regeneration from developing RPE cells. Here we report that subretinal injection of bFGF results in extensive rescue of photoreceptors in RCS rats for at least two months after the injection, and that intravitreal injection of bFGF results in even more widespread rescue, across almost the entire retina. The findings demonstrate for the first time that bFGF can act as a survival-promoting neurotrophic factor in a hereditary neuronal degeneration of the central nervous system.

• Frenkel SR, Grande DA, Collins M, Singh IJ
• Fibroblast growth factor in chick osteogenesis.
• Biomaterials. 1990; 11: 38-40
• Display abstract

• In vivo effects of FGF on osteogenesis in the chick embryo were evaluated. Day-11 embryos were injected with FGF followed by radiotracers. 3H-thymidine labelling of osteogenic cells was significantly higher following FGF administration; 3H-proline labelling over bone matrix was greater in the controls. Cartilage cells and matrix were sparsely labelled indicating a low level of metabolic activity. These data provide the first in vivo evidence that FGF stimulates osteogenic cell proliferation, while inhibiting production of bone matrix collagen. A role for FGF in embryonic osteogenesis and in fracture repair is suggested.

• Dickson C, Acland P
• Int-2: a member of the fibroblast growth factor family has different subcellular fates depending on the choice of initiation codon.
• Enzyme. 1990; 44: 225-34
• Display abstract

• The int-2 gene, which encodes a member of the fibroblast growth factor family, was discovered as a protooncogene transcriptionally activated following proviral insertion into adjacent chromosomal DNA. Analyses of the synthesis and processing of the int-2 protein, using an SV40-based vector to express cloned cDNA, showed four major products in the size range 27.5-31.5 kd that were associated with the secretory pathway. Further experiments using a cell-free translation system programmed with int-2 cRNA revealed a larger N-terminally extended protein. Site-directed mutagenesis of possible initiation codons confirmed that the first in-frame AUG codon would specify the start of a protein that includes a signal peptide for transport into the endoplasmic reticulum. However, protein synthesis also initiates at an upstream CUG codon to yield a polypeptide extended at the N-terminus by 29 amino acids. Immunofluorescent staining showed that a substantial proportion of the CUG-initiated protein resides in the cell nucleus, while a truncated int-2, lacking both the N-terminal extension and the signal peptide, was exclusively nuclear. These observations suggest that a nuclear localisation signal occurs in the body of the int-2 molecule, but is only accessible to the nuclear transport system if entry to the secretory pathway is compromised. Thus, the choice of initiation codon changes the subcellular fate of the int-2 protein and provides the potential for a duality of function through alternative transport pathways.

• Morrow NG, Kraus WE, Moore JW, Williams RS, Swain JL
• Increased expression of fibroblast growth factors in a rabbit skeletal muscle model of exercise conditioning.
• J Clin Invest. 1990; 85: 1816-20
• Display abstract

• Increased tonic contractile activity from exercise or electrical stimulation induces a variety of changes in skeletal muscle, including vascular growth, myoblast proliferation, and fast to slow fiber type conversion. Little is known about the cellular control of such changes, but pleiotropic biochemical modulators such as fibroblast growth factors (FGFs) may be involved in this response and thus may be regulated in response to such stimuli. We examined the regulation of FGF expression in an in vivo model of exercise conditioning previously shown to exhibit vascular growth and fast to slow fiber conversion. FGFs were extracted by heparin-affinity chromatography from extensor digitorum longus muscles of adult rabbits subjected to chronic motor nerve stimulation at 10 Hz. Growth factor activity (expressed in growth factor units [GFUs]) of muscle stimulated for 3 and 21 d was assayed by [3H]thymidine incorporation in 3T3 fibroblasts and compared with that present in the contralateral unstimulated muscle. A small increase in heparin-binding mitogenic activity was observed as early as 3 d of stimulation, and by 21 d mitogenic activity increased significantly when normalized to either wet weight (stimulated, 287 +/- 61 GFU/g; unstimulated, 145 +/- 39 GFU/g) or to protein (stimulated, 5.3 +/- 1.1 GFU/mg; unstimulated, 2.2 +/- 0.6 GFU/mg) (+/- SE, P less than 0.05). Western analysis demonstrated increased amounts of peptides with immunological identity to acidic and basic FGFs in stimulated muscle. The increase in FGF content observed in this study is synchronous with neovascularization, myoblast proliferation, and fast to slow fiber type conversion previously shown in this model. These results demonstrate that increased expression of FGFs is associated with motor nerve stimulation and increased tonic contractile activity of skeletal muscle, and suggests that these proteins may play a regulatory role in the cellular changes that occur during exercise conditioning.

• Furuya Y, Sato N, Watabe Y, Shimazaki J
• Effect of suramin on growth of androgen-responsive mouse tumor (Shionogi carcinoma 115) and its autonomous subline (Chiba subline 2).
• Endocrinol Jpn. 1990; 37: 933-42
• Display abstract

• Suramin has been shown to inhibit the binding of various growth factors to their receptors. Shionogi Carcinoma 115 cells (SC 115 cells) and Chiba Subline 2 cells (CS 2 cells) are clones of an androgen-responsive mouse tumor cell and its autonomous subline, respectively. Since the growth of SC 115 and CS 2 cells are assumed to be regulated by their own fibroblast growth factor (FGF)-like growth factors, the present study was undertaken to examine the effect of suramin on these cells. Suramin inhibited the growth of SC 115 and CS 2 cells in a dose dependent manner. The inhibition of suramin was reversible up to 50 micrograms/ml. Suramin reversibly changed the shape of these cells from fibroblast-like to polygonal and epithelial-like ones, and inhibited 3H-thymidine incorporation into these cells which was evoked by acidic and basic FGFs, and conditioned medium obtained from CS 2 cells. The binding of 125I-basic FGF to SC 115 and CS 2 cells was inhibited by suramin. However, suramin had no effect on growth factor production and the hst-1 gene expression on CS 2 cells. In conclusion, suramin inhibited the autocrine and paracrine growth of SC 115 and CS 2 cells by blocking the binding of autocrine growth factors to their receptors.

• Cao YH, Pettersson RF
• Human acidic fibroblast growth factor overexpressed in insect cells is not secreted into the medium.
• Growth Factors. 1990; 3: 1-13
• Display abstract

• We have overexpressed human acidic fibroblast growth factor (aFGF) in Spodoptera frugiperda Sf9 cells from a cDNA clone under the control of the promoter of the polyhedrin gene of the baculovirus Autographa californica nuclear polyhedrosis virus. A 16.5-kD product was made in recombinant virus-infected cells that specifically reacted in immunoblots with various antibodies prepared against aFGF. Recombinant aFGF was mitogenic for BHK21 cells and its activity was stimulated by heparin. The mechanism of release of FGF from mammalian cells is unknown. Both acidic and basic FGF lack classical amino-terminal signal sequences for secretion, and they are very inefficiently released from cells. Sf9 cells infected with the recombinant virus produced 10-20 mg aFGF/10(9) cells, corresponding to about 10-20 pg/cell. Despite this high level of expression, only about 0.5 and 1.3% of the total aFGF was found in the culture medium at 48 and 72 hr postinfection, respectively. This indicates that aFGF is not actively secreted out of the cells either via the normal exocytic pathway or directly through the plasma membrane in this heterologous cell system.

• Melnyk VO, Shipley GD, Sternfeld MD, Sherman L, Rosenbaum JT
• Synoviocytes synthesize, bind, and respond to basic fibroblast growth factor.
• Arthritis Rheum. 1990; 33: 493-500
• Display abstract

• Rheumatoid arthritis (RA) is a systemic disease characterized by the destructive proliferation of synovial tissue. It has been suggested that this proliferative lesion resembles a malignancy. Although polypeptide growth factors have been implicated in malignant cell growth, their role in the pathogenesis of proliferative but non-neoplastic diseases such as RA has not been extensively studied. We tested the hypothesis that the synoviocyte itself may be a source of growth factor activity. We demonstrated that culture supernatants from synoviocytes obtained from patients with RA, osteoarthritis, and traumatic joint disease contain mitogenic activity. This activity has biologic properties identical to those of basic fibroblast growth factor (bFGF). Specifically, the mitogenic activity is synergistic with insulin and binds to heparin-agarose, but elutes with 2.0M NaCl. In addition, synoviocyte extracts contain a peptide with a molecular weight of approximately 16,000, which reacts with antibody specific for bFGF. Cultured synoviocytes express the bFGF gene, express receptors for bFGF, and proliferate in response to bFGF. We conclude that bFGF derived from the synoviocytes themselves may play a role in stimulating their proliferation in an autocrine manner in disease states such as RA.

• Whitman M, Melton DA
• Signal transduction during mesoderm induction in Xenopus.
• J Reprod Fertil Suppl. 1990; 42: 249-54

• Johnson DE, Lee PL, Lu J, Williams LT
• Diverse forms of a receptor for acidic and basic fibroblast growth factors.
• Mol Cell Biol. 1990; 10: 4728-36
• Display abstract

• We recently reported the isolation of a chicken cDNA clone encoding a basic fibroblast growth factor (FGF) receptor that has three immunoglobulinlike domains in the extracellular region. We have now identified four unique human cDNA clones encoding previously unknown FGF receptor variants which contain only two immunoglobulinlike domains. Two of the human clones encode membrane-spanning receptors, and two encode putative secreted forms. Both the three- and two-immunoglobulinlike-domain forms mediate biological responsiveness to acidic and basic FGF. Thus, the first immunoglobulinlike domain of the three-domain form may have a function other than binding of acidic and basic FGF.

• Tripathi RC, Millard CB, Tripathi BJ, Chailertborisuth NS, Neely KA, Ernest JT
• Aqueous humor of cat contains fibroblast growth factor and transferrin similar to those in man.
• Exp Eye Res. 1990; 50: 109-12

• Beattie GM, Lappi DA, Baird A, Hayek A
• Selective elimination of fibroblasts from pancreatic islet monolayers by basic fibroblast growth factor-saporin mitotoxin.
• Diabetes. 1990; 39: 1002-5
• Display abstract

• Fibroblast proliferation regularly impedes the initiation and maintenance of pancreatic islet monolayers in culture. We recently characterized a specific cytotoxin to cells expressing the basic fibroblast growth factor receptor by conjugating the growth factor to saporin-6, a ribosome-inactivating protein. In contrast to untreated islets, isolated adult rat islets grown on a substrate prepared from bovine corneal endothelial cells and incubated with the mitotoxin at 10-nM concentration for 96 h were free of contaminating fibroblasts. Histological and functional studies revealed there was no damage to the islets. The results suggest that treatment of this cell type with basic fibroblast growth factor mitotoxins may be an important tool for culture of pure islets for physiological and clinical studies.

• Hondermarck H, Courty J, Ledoux D, Blanckaert V, Barritault D, Boilly B
• Evidence of high and low affinity binding sites for basic fibroblast growth factor in mouse placenta.
• Biochem Biophys Res Commun. 1990; 169: 272-81
• Display abstract

• The placenta has been shown to contain bFGF, but the presence of specific binding sites for this growth factor in this tissue remained to be established. In order to study the role of bFGF in the placenta growth, we looked for specific binding sites on mouse placental cell membranes at days 12, 14, 16, and 18 of pregnancy. At day 12, Scatchard analyses indicated that two classes of specific interaction sites for bFGF were detected. One class of high affinity binding sites was characterized by an apparent Kd of 10 pM and a binding capacity of 10 fmoles per mg of membrane protein. A second class of low affinity binding sites was detected with an apparent Kd of 60 nM and a binding capacity of 26 pmoles per mg of membrane protein. At days 14, 16 or 18, Scatchard analyses only showed low affinity binding sites with an apparent Kd of 24 nM and a binding capacity of 230 pmoles per mg of membrane protein. The characterization of these binding sites was performed by cross linking experiments that revealed two forms of specific complexes. This result suggested that the high affinity binding sites correspond to putative receptors with relative molecular masses equal to 65,000 and 85,000. The dramatic decrease of the high affinity receptor number after the 12th day of pregnancy, which is synchronous with the 9-fold increase of the low affinity binding site number, suggests that the biological activity of bFGF could be regulated by a balance between both the numbers of high and low affinity binding sites on placenta cell membranes. Thus, as it was shown for other growth factors, bFGF could only be involved at specific pregnancy stages.

• Knoerzer W, Binder HP, Schneider K, Gruss P, McCarthy JE, Risau W
• Expression of synthetic genes encoding bovine and human basic fibroblast growth factors (bFGFs) in Escherichia coli.
• Gene. 1989; 75: 21-30
• Display abstract

• Synthetic genes encoding bovine and human basic fibroblast growth factors (bFGFs) were assembled and cloned using established Escherichia coli expression plasmids. Transformed E. coli cells were able to synthesize either a fusion protein, comprising the first seven amino acids of beta-galactosidase, a linker fragment and bovine FGF, or genomic human bFGF. The two growth factors were purified from E. coli lysates by cation exchange and heparin-Sepharose affinity chromatography. The purified recombinant proteins were biologically active as monitored by their mitogenic activity for bovine aortic endothelial cells and their angiogenic capacity in the rabbit cornea.

• Rauvala H
• An 18-kd heparin-binding protein of developing brain that is distinct from fibroblast growth factors.
• EMBO J. 1989; 8: 2933-41
• Display abstract

• An 18-kd heparin-binding protein (p18) was isolated from perinatal rat brain. Although the protein closely resembles the fibroblast growth factors in its strong binding to heparin and in its apparent molecular mass, it has a distinct structure. This was concluded from the amino-terminal sequence analysis that identified a unique structure containing a cluster of lysine residues. Antipeptide antibodies were raised in rabbits according to the sequence analysis and affinity purified using a synthetic peptide. The antibodies were shown to bind specifically to p18, which was immunochemically distinct from the basic fibroblast growth factor. The antipeptide antibodies detected p18 in brain but not in liver, kidney, heart or skeletal muscle. The content of the protein was shown to undergo a remarkable developmental change corresponding to the time period of rapid sprouting of axons and dendrites in brain. The content of p18 was rapidly increased at the time of birth until the postnatal age of approximately 1 week, after which it was decreased to values less than 10% in young adults as compared to the content found in perinatal rats. p18 also enhanced neurite outgrowth in brain neurons in vitro. The protein was stained in neurons in cells dispersed from perinatal brain. The properties of p18 suggest that it has a role in the growth and maturation of brain.

• Chandrasekhar S, Harvey AK
• Induction of interleukin-1 receptors on chondrocytes by fibroblast growth factor: a possible mechanism for modulation of interleukin-1 activity.
• J Cell Physiol. 1989; 138: 236-46
• Display abstract

• Interleukin-1 is a polypeptide factor with profound effects on several cell types, such as chondrocytes, fibroblasts, and T-cells. The ability of interleukin-1 to induce the synthesis of matrix-degradative enzymes, as well as prostaglandin E2, suggests a pivotal role for this mediator in chronic inflammation. Previous studies have shown that the effect of human monocyte interleukin-1 on the synthesis of collagenase and neutral proteases by chondrocytes was enhanced by basic fibroblast growth factor. Using recombinant human interleukin-1B, we have examined whether the potentiation of interleukin-1 effects by fibroblast growth factor is related to changes in the number or affinity of interleukin-1 receptors. Our studies confirm that rabbit articular chondrocytes in culture contain a single class of high-affinity receptors for interleukin-1 with a Ka of 0.9-1.1 x 10(-13) M-1. While the untreated chondrocytes contain approximately 1,620 receptors per cell, fibroblast growth factor-treated cells exhibit a higher number of receptors (approximately 2,960 per cell) with no apparent change in the affinity. The increase in receptor number can be abolished by inhibitors of lysosomal function, indicating a requirement for intracellular processing of the fibroblast growth factor. Our results suggest that the potentiation of interleukin-1 catabolic effects by fibroblast growth factor may be related to its ability to induce additional interleukin-1 receptors on the chondrocyte cell surface.

• Loret C, Laeng P, Sensenbrenner M, Labourdette G
• Acidic and basic fibroblast growth factors similarly regulate the rate of biosynthesis of rat astroblast proteins.
• FEBS Lett. 1989; 257: 324-8
• Display abstract

• Quiescent rat astroblasts in culture have been treated for various periods of time with acidic and basic fibroblast growth factors. Both factors elicited similar effects on the cell proliferation and glutamine synthetase activity. The rate of biosynthesis of the proteins analyzed on autoradiograms of polyacrylamide gels after two-dimensional electrophoresis was also similarly modulated by the two growth factors. These results suggest that the two fibroblast growth factors act through the same membrane receptors on rat astroblasts in culture.

• Veomett G, Kuszynski C, Kazakoff P, Rizzino A
• Cell density regulates the number of cell surface receptors for fibroblast growth factor.
• Biochem Biophys Res Commun. 1989; 159: 694-700
• Display abstract

• Previous studies have shown that the binding of fibroblast growth factor to several different non-transformed cell lines decreases as cell density increases. However, it was not determined whether this is due to a reduction in receptor number or to a decrease in receptor affinity. In this study, we demonstrate that the reduction in fibroblast growth factor binding is due to a reduction in receptor number. In addition, flow cytometric analysis indicated little or no difference in the cell cycle distribution of the cells at low and high cell densities, yet the binding of fibroblast growth factor was reduced substantially at high cell densities. Thus, the reduction in growth factor binding observed in this study does not appear to result from cell cycle-dependent regulation of growth factor receptors.

• Glaser L, Wice B
• Control of differentiation in BC3H1 muscle cells.
• Environ Health Perspect. 1989; 80: 143-8
• Display abstract

• BC3H1 is a cell line that undergoes a musclelike pattern of differentiation under the appropriate conditions. We have examined the control of the synthesis of proteins characteristic of differentiated muscle in these cells as a function of their position in the cell cycle. We define two positions in the cell cycle where BC3H1 cells can remain stably quiescent. G1d is a restriction point early in the G1 portion of the cell cycle that permits the synthesis of muscle-specific proteins and is probably identical to G0. The second restriction point, G1q, occurs approximately 4 hr later in the G1 portion of the cell cycle and does not permit the synthesis of muscle-specific proteins. Movement of the cells from G1d to G1q occurs when fibroblast growth factor is added to the cells and is reversed when this growth factor is removed. Repression of the synthesis of muscle-specific proteins occurs when fibroblast growth factor is added to cells in G1d. In the case of the muscle form of creatine phosphokinase (M-CPK), the decline in the rate synthesis of this protein is a consequence of a decreased level of its mRNA. By contrast, the repression of alpha-actin synthesis, a protein synthesized only in differentiated cells, appears to be controlled at the translational level. The effect of fibroblast growth factor and other mitogens in these cells require activation of tyrosine kinase(s), but the intracellular targets of these kinases are not known. Studies by others suggest that activation of the ras oncogene can mimic the action of mitogenic polypeptides on these and other muscle cells.(ABSTRACT TRUNCATED AT 250 WORDS)

• Rifkin DB, Moscatelli D
• Recent developments in the cell biology of basic fibroblast growth factor.
• J Cell Biol. 1989; 109: 1-6

• Gospodarowicz D, Ferrara N
• Fibroblast growth factor and the control of pituitary and gonad development and function.
• J Steroid Biochem. 1989; 32: 183-91
• Display abstract

• Evidence from in vitro studies support the concept that growth factors could be involved in the development, maturation and function of endocrine organs. Included among the growth factors which are known to influence endocrine cell proliferation and differentiation is the fibroblast growth factor (FGF), which controls the proliferation, differentiation, and other functions of mesodermal- and neuroectodermal-derived cells. Its modulator, transforming growth factor beta (TGF beta), which determines the positive or negative direction of the effects of FGF, may play a role as well. In this review, we present a speculative view of how FGF in the pituitary gland, and both FGF and TGF beta in the gonads could influence the development and function of these organs through regulating mechanisms involving paracrine and autocrine control of cell proliferation and differentiation.

• Brown KD, Blakeley DM, Brigstock DR
• Stimulation of polyphosphoinositide hydrolysis in Swiss 3T3 cells by recombinant fibroblast growth factors.
• FEBS Lett. 1989; 247: 227-31
• Display abstract

• Mitogenic concentrations of recombinant acidic or basic fibroblast growth factor (FGF) stimulated the accumulation of [3H]inositol phosphates ([3H]IPs) in Swiss 3T3 cells pre-labelled for 48 h with [3H]inositol. Maximal effects were obtained at 0.3 ng/ml and 3 ng/ml for basic and acidic FGF, respectively. Higher doses of either factor led to a diminished stimulation. FGF also stimulated 45Ca2+ release from cells pre-labelled with the isotope. However, FGF-stimulated production of [3H]IPs and release of 45Ca2+ exhibited marked differences when compared with the responses to the peptide mitogen bombesin; the FGF responses were markedly slower and were not inhibited by phorbol esters.

• Burgess WH, Maciag T
• The heparin-binding (fibroblast) growth factor family of proteins.
• Annu Rev Biochem. 1989; 58: 575-606

• Mascarelli F, Raulais D, Courtois Y
• Fibroblast growth factor phosphorylation and receptors in rod outer segments.
• EMBO J. 1989; 8: 2265-73
• Display abstract

• Acidic and basic fibroblast growth factors (aFGF and bFGF) have been isolated and purified from rod outer segments (ROS). aFGF is tightly bound to ROS membranes and can be specifically released by ATP. We show that this mechanism is dependent on the phosphorylation of aFGF itself. Phorbol 12-myristate 13-acetate (PMA) enhances this phenomenon independently of rhodopsin phosphorylation. This demonstrates that aFGF release from ROS membranes is dependent on its phosphorylation by endogenous kinase C. In addition specific binding sites for exogenous FGFs have been identified on ROS and disc membranes. A single high affinity site with a Kd of 40 pM was present in intact ROS while an additional low affinity site with a Kd of 300-600 pM was present in leaky ROS or in disc membranes. Light or ATP modified neither these Kd nor the apparent number of sites. The presence of specific receptors for FGFs and the kinase C dependent release of endogenous membrane bound aFGF suggest an autocrine mechanism which may be involved in photoreceptor cell biology.

• Klagsbrun M
• The fibroblast growth factor family: structural and biological properties.
• Prog Growth Factor Res. 1989; 1: 207-35
• Display abstract

• This article summarizes the structural and biological properties of the family of fibroblast growth factors (FGF). Basic FGF (bFGF) and acidic FGF (aFGF) are the best characterized members of this family. bFGF and aFGF are potent modulators of cell proliferation, motility and differentiation. They are also potent angiogenesis factors in vivo. Some of the important biological characteristics of bFGF and aFGF discussed in the review include the affinity of bFGF and aFGF for heparin, their lack of secretion in culture and their association with extracellular matrix. Recently, several oncogenes, 40-50% homologous in sequence to bFGF and aFGF have been identified. These include int-2, hst, K-fgf and FGF-5. The structural and biological properties of these FGF-related oncogenes are also discussed.

• Weiner HL, Swain JL
• Acidic fibroblast growth factor mRNA is expressed by cardiac myocytes in culture and the protein is localized to the extracellular matrix.
• Proc Natl Acad Sci U S A. 1989; 86: 2683-7
• Display abstract

• Acidic and basic fibroblast growth factors are heparin-binding proteins that induce cellular proliferation, mesodermal development, and vascular growth. As such, they may be important in cardiac development and disease. To determine whether cardiac myocytes contain fibroblast growth factors, neonatal rat cardiac myocytes were studied in primary culture and compared to primary cultures of nonmyocyte cardiac cells. Northern blot analysis revealed a 4.0-kilobase mRNA in myocytes that hybridized to acidic fibroblast growth factor cDNA and was not detectable in nonmyocyte cultures. Western blot analysis demonstrated the accumulation of a 15-kDa peptide with immunological identity to acidic fibroblast growth factor in extracts of extracellular matrix from myocyte cultures that was not detectable in similar extracts of nonmyocyte extracellular matrix. No acidic fibroblast growth factor-like protein was detectable in cellular lysates from either myocyte or nonmyocyte cultures. These results demonstrate that neonatal cardiac myocytes express acidic fibroblast growth factor mRNA and deposit a protein with immunological identity to acidic fibroblast growth factor into the extracellular matrix. The results suggest that acidic fibroblast growth factor produced by cardiac myocytes may mediate, through both paracrine and autocrine mechanisms, such diverse processes as myocyte differentiation, cellular proliferation, and vascular growth in the heart.

• Besnard F, Perraud F, Sensenbrenner M, Labourdette G
• Effects of acidic and basic fibroblast growth factors on proliferation and maturation of cultured rat oligodendrocytes.
• Int J Dev Neurosci. 1989; 7: 401-9
• Display abstract

• A pure culture of oligodendrocytes has been developed starting from brain hemispheres of newborn rats. Various effects of acidic and basic fibroblast growth factors (FGFs) on the development of oligodendrocytes have been examined and compared. Both factors elicited similar effects, i.e. stimulation of the proliferation, inhibition of the specific activity of the marker enzyme 2',3'-cyclic nucleotide 3'-phosphodiesterase and decrease of the ratio of myelin basic protein positive cells. These results indicate that FGFs are very potent mitogens for oligodendrocytes, even in the absence of other cell types, but that they elicit a negative effect on the cell maturation, possibly related to their strong effect on proliferation.

• Gospodarowicz D
• Expression and control of vascular endothelial cells: proliferation and differentiation by fibroblast growth factors.
• J Invest Dermatol. 1989; 93: 3947-3947
• Display abstract

• Evidence from in vitro studies supports the concept that growth factors could be involved in the development and function of the vascular tree. Among the growth factors known to influence vascular endothelial cells' proliferation and differentiation, in vitro, are the fibroblast growth factors. These consist of two closely, structurally, related polypeptides that differ by their isoelectric point and have been called basic and acidic fibroblast growth factor. These growth factors, in particular basic fibroblast growth factor, which is expressed and synthesized by vascular endothelial cells, could influence the development, remodeling, and function of the vascular tree through regulating mechanisms involving paracrine and autocrine control of cell proliferation and differentiation.

• Rodan SB, Wesolowski G, Thomas KA, Yoon K, Rodan GA
• Effects of acidic and basic fibroblast growth factors on osteoblastic cells.
• Connect Tissue Res. 1989; 20: 283-8
• Display abstract

• Acidic (a) and basic (b) fibroblast growth factors (FGFs) are two related mitogenic and angiogenic factors. They are multifunctional in that they can affect proliferation and induce or delay differentiation. Both aFGF and bFGF were shown to stimulate proliferation of calvaria cells in situ as well as osteoblast-enriched calvaria-derived cells. bFGF was also found to suppress the expression of alkaline phosphatase, parathyroid hormone stimulatable adenylate cyclase, osteocalcin, and type I collagen in the osteoblastic ROS 17/2.8 cells. To explore a possible role for guanine nucleotide binding proteins we assessed the effects of pertussis toxin (PT) on FGF action. PT had opposite effects to those of bFGF on all parameters examined.

• Slack JM
• Peptide regulatory factors in embryonic development.
• Lancet. 1989; 1: 1312-5

• Mereau A, Pieri I, Gamby C, Courty J, Barritault D
• Purification of basic fibroblast growth factor receptors from bovine brain.
• Biochimie. 1989; 71: 865-71
• Display abstract

• Fibroblast growth factors are proteins which play a major role, in vitro and in vivo, in the control of cellular growth and differentiation of a large number of cells. Biological activities of these factors are mediated by the interaction with specific membrane receptors. Previous studies indicated that the apparent molecular weight of a family of these receptors for the basic form of Fibroblast Growth Factor (bFGF), ranges from 125 to 165 kDa according to cell species and types. We have purified this family of receptors from bovine brain. We first set up a radioreceptor assay to detect receptors throughout the purification by measuring its ability to inhibit the fixation of radiolabeled bFGF to insolubilized membranes from bovine brain. The purification was also monitored by using cross-linking reagents in order to allow the visualization of radiolabeled bFGF bound to its receptor. The first purification steps involved 2 anion-exchange chromatographic steps, DEAE Trysacryl and FPLC Mono Q, and yielded an enrichment over 500 fold. Affinity chromatography with bFGF immobilized on Sepharose 4B was then performed. Covalent fixation of bFGF to the Sepharose matrix was carried out in presence of N-acetylated heparin in order to protect the recognition site for bFGF on its receptor. These 3 chromatographic steps yielded only 2 bands of apparent molecular weight of 100 kDa and 135 kDa as detected by electrophoresis. These 2 bands are also detected after chromatography on immobilized wheat germ agglutinin hence confirming the presence of carbohydrates on bFGF receptors.

• Klagsbrun M, Edelman ER
• Biological and biochemical properties of fibroblast growth factors. Implications for the pathogenesis of atherosclerosis.
• Arteriosclerosis. 1989; 9: 269-78

• Ruta M et al.
• Receptor for acidic fibroblast growth factor is related to the tyrosine kinase encoded by the fms-like gene (FLG).
• Proc Natl Acad Sci U S A. 1989; 86: 8722-6
• Display abstract

• We have previously isolated a human gene from an endothelial cell cDNA library encoding a putative tyrosine kinase; we have designated this gene the fms-like gene (FLG). To analyze the gene product(s) of FLG, we have generated rabbit polyclonal antibodies directed against a synthetic peptide from FLG and used it to immunoprecipitate biosynthetically labeled FLG protein from a variety of human cell lines. These antibodies specifically recognized glycoprotein(s) of 100, 120, and 135 kDa with protein cores of 90 and 110 kDa. Acidic fibroblast growth factor (aFGF) stimulated tyrosine kinase activity of FLG in vitro and in living cells, suggesting that FLG encodes the membrane receptor for aFGF. Further supporting evidence came from cross-linking experiments on intact cells with the covalent cross-linking agent disuccinimidyl suberate and 125I-labeled aFGF as a specific probe. The cross-linked 125I-labeled aFGF-aFGF receptor complex was specifically immunoprecipitated with FLG antipeptide antibodies. It appears, therefore, that the receptor(s) for aFGF is related to the FLG gene product.

• Speir E, Zhou YF, Lee M, Shrivastav S, Casscells W
• Fibroblast growth factors are present in adult cardiac myocytes, in vivo [corrected and issued with original paging in Biochem Biophys Res Commun 1988 Dec 30;157(3)]
• Biochem Biophys Res Commun. 1989; 159: 1336-40
• Display abstract

• We have previously shown that the adult heart contains mitogens immunologically identical to acidic and basic fibroblast growth factors. To determine whether these proteins are present in myocytes, we subjected lysates of freshly isolated myocytes to heparin-affinity chromatography. The 1.1 M - 3 M NaCl eluates stimulated incorporation of thymidine into DNA in quiescent Balb/c 3T3 fibroblasts, caused proliferation of vascular endothelial cells, and cross-reacted with antisera raised against acidic (1.1 M) and basic FGF's (1.5 M) by Western blotting and by RIA. These proteins may be involved in cellular differentiation and proliferation and may play an important role in regenerative and repair processes in the heart.

• Smith BT, Post M
• Fibroblast-pneumonocyte factor.
• Am J Physiol. 1989; 257: 1748-1748
• Display abstract

• Fibroblast-pneumonocyte factor (FPF) is an organ-specific differentiation factor produced, under glucocorticoid regulation, by the fetal lung fibroblast, which in turn enhances the differentiation of the alveolar type II cell with respect to surfactant production. In this article we review the appearance of the type II cell phenotype during fetal life and the action of FPF to time full acquisition of this phenotype. Evidence supporting a physiological role for this factor is reviewed as are details of its production by the fetal lung fibroblast and action on the type II cell. Finally we summarize possible future clinical advantages of using such a material to regulate lung maturation compared with currently used maternal glucocorticoid therapy. These include a more rapid effect and bypassing the inhibition of glucocorticoid action seen in male fetuses and in infants of diabetic mothers.

• Genot E, Petit-Koskas E, Sensenbrenner M, Labourdette G, Kolb JP
• Potentiation of the proliferative response of human B lymphocytes to low molecular weight B cell growth factor (LMW-BCGF) by fibroblast growth factors (FGFs).
• Cell Immunol. 1989; 122: 424-39
• Display abstract

• Both acidic and basic fibroblast growth factor (FGF), although devoid alone of growth-promoting ability on resting or activated human lymphoid B cells, were found to markedly increase the proliferative response of anti-mu-chain or SAC preactivated B cell blasts to the low molecular weight B cell growth factor (LMW-BCGF) and to enhance the costimulatory response of resting B cells to anti-mu-chain and LMW-BCGF. This potentiating effect was also observed for a LMW-BCGF-dependent B cell tumor derived from a lymphocytic nodular lymphoma. Other growth factors acting on fibroblasts, such as epidermal growth factor, alpha-thrombin, platelet-derived growth factor, and insulin-like growth factor-I did not display such enhancing effect on LMW-BCGF-driven proliferation. Activated, but not resting B cells were found to bear receptor sites for FGFs and from kinetics experiments, it is suggested that LMW-BCGF induces competence expression for FGFs in those cells. Moreover, the LMW-BCGF-elicited generation of inositoltrisphosphate resulting from polyphosphoinositides hydrolysis was increased in the presence of FGF.

• Lappi DA, Martineau D, Baird A
• Biological and chemical characterization of basic FGF-saporin mitotoxin.
• Biochem Biophys Res Commun. 1989; 160: 917-23
• Display abstract

• Basic fibroblast growth factor (FGF) and saporin-6, a ribosome-inactivating protein, were chemically conjugated and characterized as a cytotoxin to cells expressing the basic FGF receptor. Structural and Western blot analysis of the conjugate showed that it contained saporin and basic FGF in equimolar amounts. The conjugate inhibited protein synthesis in a cell-free system and had potent cytotoxic activity (ID50 = 25pM) for cells expressing the basic FGF receptor. It is equipotent with basic FGF in radioreceptor assays and elutes from heparin Sepharose columns with 2M NaCl. The activity of the mitotoxin can be inhibited by competition with an excess of basic FGF but not nerve growth factor. The possibility that this mitotoxin can be used as an anti-angiogenic factor in paradigms that involve basic FGF is discussed.

• Begin R, Dufresne A, Cantin A, Masse S, Sebastien P, Perrault G
• Carborundum pneumoconiosis. Fibers in the mineral activate macrophages to produce fibroblast growth factors and sustain the chronic inflammatory disease.
• Chest. 1989; 95: 842-9
• Display abstract

• Carborundum is a synthetic abrasive manufactured through fusion of high grade silica sand and finely ground carbon in an electric furnace at 2,400 degrees C. It had been considered an inert dust until recently. Two recent epidemiologic studies in Quebec have documented an excess of interstitial lung disease in plant workers and some 30 workers have received workman compensation. Histopathologic lesions have been described in four of the workers. To further investigate the carborundum pneumoconiosis, nine groups of eight sheep were exposed once in the tracheal lobe to either 100 ml saline, 100 mg latex beads in 100 ml saline, 100 mg graphite in 100 ml saline, 100 mg carborundum particles in 100 ml saline, 100 mg ashed carborundum particles in 100 ml saline, 100 mg of quartz (Minusil-5) in 100 ml saline, 100 mg crocidolite fibers in 100 ml saline, 100 mg carborundum fibers in 100 ml saline, and 100 mg ashed carborundum fibers in 100 ml saline solutions. The animals had BAL at two-month intervals and autopsy at month 8. The BAL analyses of cellularity, cytotoxicity and fibrogenicity, in association to necropsy histopathology, documented that all particles except for quartz were inert. The two-carborundum fiber samples produced a similar sustained nodular fibrosing alveolitis and crocidolite asbestos fibers produced a peribronchiolar fibrosing alveolitis of comparable severity. Thus, the major bioactive dusts in the carborundum manufacturing process are quartz particles and the carborundum fibers generated in the process. The latter have fibrogenic activities comparable to asbestos fibers of similar size and are likely to contribute to the pathogenesis of the interstitial lung disease of carborundum workers.

• Blanquet PR, Patte C, Fayein N, Courtois Y
• Identification and isolation from bovine epithelial lens cells of two basic fibroblast growth factor receptors that possess bFGF-enhanced phosphorylation activities.
• Biochem Biophys Res Commun. 1989; 160: 1124-31
• Display abstract

• Cross-linking of bound 125I-basic fibroblast growth factor (bFGF) to bovine epithelial lens cells identified two labelled species whose apparent molecular weights were identical with those of two phosphorylated proteins. The bFGF-stimulated phosphorylation of these proteins was shown to be rapid, suggesting an autophosphorylation process. To demonstrate that the phosphorylated proteins were indeed the bFGF-binding molecules, the two components were purified to homogeneity and their bFGF-binding activity was examined. We conclude that bFGF stimulates the phosphorylation of two receptors of 130 and 160 kDa in bovine epithelial lens cells.

• Hanai K et al.
• Central action of acidic fibroblast growth factor in feeding regulation.
• Am J Physiol. 1989; 256: 21723-21723
• Display abstract

• A phasic increase in activity similar to that of acidic fibroblast growth factor (aFGF) was detected in the cerebrospinal fluid of rats after feeding or after an intraperitoneal injection of glucose. This FGF-like activity in the cerebrospinal fluid was bioassayed in two systems: depression of the feeding response of Hydra and DNA synthesis-stimulating activity in BALB/c 3T3 cells after fractionation on a heparin affinity column. Dynamic feeding-related changes in activity of aFGF, basic FGF, and other growth factors were detected by both bioassay systems. Intracerebroventricular microinfusion of aFGF suppressed food intake in rats. Central infusion of inactivated aFGF, or peripheral administration of aFGF, in doses equivalent to or higher than those administered centrally was without effect. Electrophoretically applied aFGF specifically suppressed the activity of glucose-sensitive neurons in the lateral hypothalamus. Glucose-insensitive neurons were only slightly affected. The results suggest that aFGF may participate in the regulation of feeding at the level of the central nervous system.

• Aspenberg P, Lohmander LS
• Fibroblast growth factor stimulates bone formation. Bone induction studied in rats.
• Acta Orthop Scand. 1989; 60: 473-6
• Display abstract

• Implantation of demineralized bone matrix in rodents elicits a series of cellular events leading to the formation of new bone inside and adjacent to the implant. This process is believed to be initiated by an inductive protein present in bone matrix. It has been suggested that local growth factors may further regulate the process once it has been initiated. This investigation was designed to study the effect of adding a growth factor to the inductive implant. Pairs of demineralized rat femoral diaphyses were implanted intramuscularly in rats. The marrow canal of one implant in each pair was filled with a carboxymethyl cellulose gel containing 75 ng of recombinant human basic fibroblast growth factor (FGF). The other implant in each pair served as a control. It was either filled with the gel without FGF or left untreated. Bone formation was induced by all the implants after 3 weeks. The amount of mineralized tissue in the FGF-treated implants was 25 percent greater than in untreated controls. The carboxymethyl cellulose gel alone did not affect the bone yield.

• Wu KF
• [Significance and modulation of production of tumor growth inhibitor derived from human embryonic myofibroblasts]
• Zhonghua Zhong Liu Za Zhi. 1989; 11: 419-21
• Display abstract

• In this report, the production and modulation of tumor growth inhibitor (HFDI) derived from human embryonic myofibroblasts were studied. HFDI was obtained from 3/30 cultures of human embryonic myofibroblasts, which was obviously lower than the incidences of Australia Caucasian's myofibroblast (6/6) and adult's skin fibroblast (4/6). It is suggested that the inheritance play certain role in HFDI production. Calf serum had a significant effect on HFDI production, though without any linear relationship. It seems that HFDI is not derived from calf serum but the factors influencing the production of HFDI are in the calf serum. HFDI could be induced by inactivated BCG from non- or low-producing myofibroblasts. It is indicated that the production of HFDI is related to bacterial contamination (in vitro) or infection (in vivo). The hypothesis is proposed that tumor inhibitors be induced by bacterial infection from the fibroblasts. HFDI-like activity was identified in 4 healthy donors' sera suggesting that HFDI could enter the blood.

• Gillespie LL, Paterno GD, Slack JM
• Analysis of competence: receptors for fibroblast growth factor in early Xenopus embryos.
• Development. 1989; 106: 203-8
• Display abstract

• Xenopus ectodermal cells have previously been shown to respond to acidic and basic FGF by differentiating into mesodermal tissue. In the present study, ectodermal explants from Xenopus blastulae were shown to have high affinity binding sites for 125I-aFGF (Kd = 1.4 X 10(-10) M). The total number of sites, determined by Scatchard analysis, was 3 X 10(8) per explant (surface area of approximately 1 mm2). Two putative receptors of relative molecular mass 130,000 and 140,000 were identified by chemical crosslinking to 125I-aFGF. Both acidic and basic FGF, but not TGF beta 2, could compete for affinity labelling of these bands. The receptor density at the cell surface parallels the developmental competence of Xenopus animal pole cells to respond to FGF. Receptors are present at highest density in the marginal zone but are not restricted to cells in this region.

• Shing Y
• Heparin-copper biaffinity chromatography of fibroblast growth factors.
• J Biol Chem. 1988; 263: 9059-62
• Display abstract

• A novel method is described to separate and identify the various forms of fibroblast growth factor (FGF) based on their differential affinities for both heparin and copper. FGFs were extracted from bovine hypothalamus and purified by batchwise adsorption to heparin-Sepharose. The partially purified FGFs were then applied to an affinity column prepared by mixing equal portions of heparin-Sepharose and copper-Sepharose. The column was rinsed consecutively with the following four reagents: (i) 2 M NaCl, (ii) 0.6 M NaCl, (iii) 0.6 M NaCl plus 10 mM imidazole, and (iv) 0.6 m NaCl. FGFs were then eluted with a linear NaCl/imidazole gradient (from 0.6 m NaCl without imidazole to 2 M NaCl plus 10 mM imidazole). Fractions eluted from the column were analyzed by sodium dodecyl sulfate-gel electrophoresis with silver staining and electrophoretic immunoblot using site-specific antibodies against basic and acidic FGF. The results demonstrate that it is possible to resolve from hypothalamus at least two basic FGF species (with Mr values of 19,000 and 18,000) and three acidic FGF species (with Mr values of 18,000, 16,400, and 15,600). These findings indicate that heparin-copper biaffinity chromatography may have wide applicability in the study of the structure and activity of FGFs.

• Austgulen R, Nissen-Meyer J
• A physico-chemical comparison of the monocyte-derived fibroblast growth factor and the tumour necrosis factor.
• APMIS. 1988; 96: 352-6
• Display abstract

• The chromatographic characteristics of the human monocyte-derived growth stimulatory activity towards diploid FS-4 human fibroblasts has been studied. The fibroblast growth stimulatory factor had an apparent isoelectric point of 5.8 as determined by chromatofocusing, and a molecular weight in the range 70,000-30,000 as determined by gel filtration. The growth stimulatory factor eluted together with the tumour necrosis factor (TNF) upon cation exchange chromatography, chromatofocusing and gel filtration. Moreover, the fibroblast growth stimulatory activity in crude monocyte supernatants as well as the activity in the gel filtration column fractions was neutralized by antiserum raised against recombinant TNF (rTNF). The results indicate that the monocyte-derived fibroblast growth stimulatory activity is largely due to TNF.

• Sharma A, Dahiya R
• Bovine basic fibroblast growth factor: identification and binding of its receptor on PC12 cells.
• Biomed Biochim Acta. 1988; 47: 975-83
• Display abstract

• Basic fibroblast growth factor (FGF) was purified to homogeneity from bovine pituitary glands on the basis of its ability to stimulate Swiss 3T3 cell proliferation. The binding characteristics of biologically active 125[I]-FGF to PC12 cells have been studied. Binding of 125[I]-FGF was found to be specific, as the bound 125[I]-FGF was displaced by native FGF but not by epidermal growth factor (EGF) or nerve growth factor (NGF). Binding was saturable both as a function of time and of the concentration of 125[I]-FGF. Scatchard analysis of the binding data revealed the presence of 1.05.10(5) binding sites/cell with an apparent KD of 258 pM. When 125[I]-FGF binding was studied in PC12 cell membranes, it was found that the membraneous binding site displayed a lower KD of 28 pM. 125[I]-FGF was covalently cross-linked to its cell surface receptor on intact PC12 cells using the homobifunctional agent disuccinimidyl suberate. The covalently cross-linked 125[I]-FGF identified a single detergent soluble FGF binding site with an apparent molecular weight of 145,000 by SDS-PAGE analysis. Unlabeled FGF was found to compete with 125[I]-FGF for binding to the same receptor site. No labelling was observed in the absence of the cross-linker or when heat inactivated 125[I]-FGF was used. In conclusion, the present studies demonstrate for the first time the presence of FGF receptor on PC12 cells.

• Feige JJ, Baird A
• Glycosylation of the basic fibroblast growth factor receptor. The contribution of carbohydrate to receptor function.
• J Biol Chem. 1988; 263: 14023-9
• Display abstract

• We have examined the glycosylation of the basic fibroblast growth factor (bFGF) receptor to determine whether carbohydrates contribute to receptor structure and function. Using a combination of cross-linking and radioreceptor assays, we demonstrated that the two bFGF receptors in baby hamster kidney cells have protein cores of 100 and 125 kDa. They are glycosylated to high mannose forms of 115 and 140 kDa and further processed to their mature forms of 130 and 150 kDa. Because peptide:N-glycosidase F, but not endo-alpha-N-acetylgalactosamidase can reduce the size of the bFGF receptors, the carbohydrate residues of the receptor appear all N-linked. The inability of deglycosylated receptors to bind 125I-bFGF supports the notion that the carbohydrate residues are required for receptor function. Furthermore, the capacity of the wheat germ agglutinin lectin to inhibit 125I-bFGF binding and the biological activity of bFGF suggests that N-acetylglucosamine residues are functionally significant components of the receptor.

• Schweigerer L
• Basic fibroblast growth factor as a wound healing hormone.
• Trends Pharmacol Sci. 1988; 9: 427-8

• de Lapeyriere O, Raybaud F, Mattei MG, Birnbaum D
• [FGF gene family and their role in human tumors]
• Pathol Biol (Paris). 1988; 36: 863-5

• Woodland H, Jones L
• Growth factors in amphibian cell differentiation.
• Nature. 1988; 332: 113-5

• Thomas KA
• Transforming potential of fibroblast growth factor genes.
• Trends Biochem Sci. 1988; 13: 327-8

• Matuo Y et al.
• Stabilization of fibroblast growth factors by a non-cytotoxic zwitterionic detergent, 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate (CHAPS).
• In Vitro Cell Dev Biol. 1988; 24: 477-80
• Display abstract

• The potential usefulness of a zwitterionic detergent, 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate (CHPAS), in the stabilization of acidic and basic fibroblast growth factors (FGFs) was examined. Among several detergents, CHAPS was found to be not only non-cytotoxic but also most useful in handling the diluted preparations of FGFs. The advantages are as follows: 1) at lower concentrations than 0.01% CHAPS did not affect growth factor activity of calf serum (CS) and the growth rate of BALB/c 3T3 cells. The primary culture of rat prostate epithelium and colony formation of NRK-49F cells were hardly influenced by CHAPS lower than 0.003%; 2) the loss of FGFs that usually occurs due to their adherence to the surface of storage containers was effectively prevented by inclusion of 0.1% CHAPS; 3) the recovery of FGFs after storage or dialysis was significantly enhanced by inclusion of 0.1% CHAPS; 4) CHAPS at lower concentrations than 0.1% does not interfere with amino acid analysis, except that Thr may be misled only when the ratio of protein/CHAPS is low; 5) amino acid sequence analysis was hardly disturbed by CHAPS up to 0.5%. These results indicate that CHAPS is useful as a stabilizing agent for various kinds of polypeptides capable of showing biological activity at a low concentration.

• Hicks D, Courtois Y
• Acidic fibroblast growth factor stimulates opsin levels in retinal photoreceptor cells in vitro.
• FEBS Lett. 1988; 234: 475-9
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• It is demonstrated that newborn rat retinal photoreceptor cells can differentiate in monolayer culture, and synthesize de novo photoreceptor-specific proteins such as opsin. When maintained in serum supplemented medium on a laminin substrate, these cells survive for up to 3 weeks. The addition of acidic fibroblast growth factor stimulates an increase in the levels of opsin of 5-10-fold control values, and prolongs cell survival by up to 6 days.

• Gospodarowicz D
• Molecular and developmental biology aspects of fibroblast growth factor.
• Adv Exp Med Biol. 1988; 234: 23-39

• Walicke PA, Baird A
• Trophic effects of fibroblast growth factor on neural tissue.
• Prog Brain Res. 1988; 78: 333-8

• Haynes LW
• Fibroblast (heparin-binding) growing factors in neuronal development and repair.
• Mol Neurobiol. 1988; 2: 263-89
• Display abstract

• Nearly thirty growth and trophic factors that have been purified from mammalian tissues in the last 15 yr have been found to share chemical identity. The results of their chemical purification and molecular cloning show that they are two distinct polypeptides (Mr 17,400 and 18,400), each of which gives rise to families of smaller size peptides. These peptides share a common affinity for heparin. In view of this property, a common nomenclature for the two principle peptide growth factors (heparin-binding growth factor classes 1 and 2; HBGF-1 and -2) has been proposed. However, the names acidic and basic Fibroblast Growth Factors (aFGF,bFGF), which were applied to them originally to describe their mitogenic activity, are more commonly in use and will therefore be adopted in this review. Brain tissue is one of the richest sources of FGFs. It has been used as a starting point for their chemical purification and to prepare genomic libraries for molecular cloning of the aFGF and bFGF genes. There is increasing evidence that these growth factors, expressed in neurons and glia throughout the mammalian nervous system, are implicated in neuronal cell proliferation, differentiation, and histogenesis. FGFs have a strong affinity not only for heparin, but also for the related heparan sulphate proteoglycans that are abundant in neural tissues. This fact provides a clue to the importance of tissue-associated proteoglycans in mediating the release, sequestration, and activation of FGFs and the modulation of their receptor binding and bioactivity. The relevance of FGFs to neural development and their mechanisms of action in neurons will be considered in light of the existing literature describing their biological properties and activity in mesodermal cell types. Evidence is reviewed showing that FGFs have in vivo biological activity, ameliorating the degeneration of central and peripheral neurons after axotomy. The presence and implications of high levels of FGFs in adult mammalian brain provides a direction for future research into neural regeneration. The bioactivity of FGFs in neural tissue may not depend on the regulation of their expression per se, but on the subregional modification of their interaction with proteoglycans.

• Biswas SB, Hammond RW, Anderson LD
• Fibroblast growth factors from bovine pituitary and human placenta and their functions in the maturation of porcine granulosa cells in vitro.
• Endocrinology. 1988; 123: 559-66
• Display abstract

• Both acidic and basic fibroblast growth factors (FGFs) are present in pituitary, as demonstrated by Affigel-heparin affinity chromatography and immunoblot analysis. Unlike in the pituitary, only basic FGF appears to be present in human placenta. The purification of these growth factors from bovine pituitary and human placenta has been described. Immunoblot analysis using polyclonal antibodies specific for either basic FGF or both acidic and basic FGF have verified the results of purification. We have determined a novel function of acidic and basic FGF in the maturation of porcine granulosa cells in vitro. Basic FGF (1.0 ng/ml), purified from bovine pituitary functioned as a potent inhibitor of LH/hCG receptor induction (91 +/- 1.2%; P less than 0.0001), and progesterone secretion (45 +/- 2.9%; P less than 0.0001), two key steps in the maturation of granulosa cells. Human placental basic FGF demonstrated comparable inhibition characteristics. Bovine pituitary acidic FGF (50 ng/ml) appear to function as a relatively weak inhibitor (63.5 +/- 5.1% for LH/hCG receptor induction; 50 +/- 1.8% for progesterone secretion). Consequently, our analysis indicates that FGF may play an important role(s) in the maturation of ovarian granulosa cells.

• Rizzino A, Kuszynski C, Ruff E, Tiesman J
• Production and utilization of growth factors related to fibroblast growth factor by embryonal carcinoma cells and their differentiated cells.
• Dev Biol. 1988; 129: 61-71
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• Previous studies have established that embryonal carcinoma (EC) cells produce several different growth factors, but express few, if any, receptors for epidermal growth factor, platelet-derived growth factor, or transforming growth factor type-beta. In this study, the production and utilization of fibroblast growth factor (FGF) by EC cells and their differentiated cells were investigated. We have determined that EC cells produce a heat-labile, heparin-binding factor that competes with FGF for binding to membrane receptors and appears to be immunologically related to FGF. The same or a similar factor is produced by three different EC cell lines, including a multipotent human EC cell line. However, production of this factor is apparently reduced when each EC cell line differentiates. Unlike the parental EC cells, the differentiated cells respond to FGF by growth stimulation and the growth responses to FGF correlate with increased binding of FGF. Although the binding data indicate that both the EC cells and their differentiated cells exhibit high affinity receptors for FGF, the differentiated cells express these receptors at levels approximately 10-fold higher. These findings suggest that the FGF-related growth factor could influence the growth of EC cells or their differentiated cells.

• Seed J, Olwin BB, Hauschka SD
• Fibroblast growth factor levels in the whole embryo and limb bud during chick development.
• Dev Biol. 1988; 128: 50-7
• Display abstract

• A growth factor with properties very similar to fibroblast growth factor (FGF) was detected in the yolk and white of unfertilized chick eggs, and in the limb bud and bodies of Day 2.5 (stage 18)-13 chick embryos using two complementary and highly sensitive biological assays-competition of 125I-a-FGF binding to the FGF receptors of 3T3 cells and stimulation of DNA synthesis in MM14 cells, a permanent mouse skeletal muscle cell line that is dependent upon FGF for proliferation. Further evidence of the similarity of this growth factor to FGF is provided by the finding that biological activity is lost when the material is bound to a heparin-Sepharose column and restored upon elution with 2.5 M NaCl; the 2.5 M NaCl fraction from Day 12 embryos contains several polypeptides of apparent molecular weights 12,500-17,500. The level of FGF in the embryonic chick body is fairly constant between Days 2.5 and 6 (stages 18-29), ranging between 1 and 2 ng FGF/mg protein; but thereafter the level increases so that by Day 13 the body contains about 15 ng FGF/mg protein. In contrast, the level of FGF in the limb but is higher than that in the rest of the body until Day 5 (stage 27); it then undergoes a transient decrease between Days 6 and 7, after which it increases but remains below the level observed in the remainder of the body.

• Tripathi RC, Millard CB, Tripathi BJ, Reddy V
• A molecule resembling fibroblast growth factor in aqueous humor.
• Am J Ophthalmol. 1988; 106: 230-1

• Liu LM, Nicoll CS
• Evidence for a role of basic fibroblast growth factor in rat embryonic growth and differentiation.
• Endocrinology. 1988; 123: 2027-31
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• The possibility that basic fibroblast growth factor (FGF) plays a role in the regulation of mammalian embryogenesis was investigated. When transplanted under the renal capsule of syngeneic juvenile hosts, 10-day-old rat embryos grow rapidly, and tissue differentiation occurs in an essentially normal manner. Infusion of antiserum to bovine FGF into the renal artery of kidneys bearing such transplants significantly retarded their growth, and differentiation of all tissues of endodermal and some of those of mesodermal origin was completely suppressed; differentiation of other tissues of mesodermal origin was partially inhibited. Infusion of nonimmune serum had no such effects. Infusion of recombinant bovine FGF into the renal artery stimulated growth of embryo transplants on the infused kidney, but not of those under the renal capsule of the contralateral kidney, and it did not affect growth of the infused kidney itself. These results suggest that FGF may be important for growth and differentiation of endodermal and some mesodermal tissues of mammalian embryos.

• Perraud F, Besnard F, Pettmann B, Sensenbrenner M, Labourdette G
• Effects of acidic and basic fibroblast growth factors (aFGF and bFGF) on the proliferation and the glutamine synthetase expression of rat astroblasts in culture.
• Glia. 1988; 1: 124-31
• Display abstract

• The two fibroblast growth factors called acidic and basic FGF (aFGF and bFGF) show a strong homology (55%) of their amino acid sequence (Esch et al.: Proc. Nat. Acad. Sci. USA 85:6507-6511, 1985). The effects of these factors on the rate of proliferation of rat astroblasts and on the expression of glutamine synthetase activity in cells grown in primary culture were investigated and compared under various culture conditions. In all the experimental conditions used, both growth factors triggered the proliferation of the cells to the same extent and with similar dose dependence. The mitogenic activities of aFGF and bFGF were potentiated similarly by heparan sulfate and by heparin, with a maximum stimulation of about 100% at 100 micrograms/ml heparin. Treatment of the cells with either of the two factors resulted in identical enhancement of the activity of glutamine synthetase relative to total proteins. These results suggest that both factors act either through the same membrane receptors or through different receptors that mediate nearly identical effects.

• Floros J, Nielsen HC, Torday JS
• Dihydrotestosterone blocks fetal lung fibroblast-pneumonocyte factor at a pretranslational level.
• J Biol Chem. 1987; 262: 13592-8
• Display abstract

• Fibroblast pneumonocyte factor (FPF) synthesis by fetal rat lung fibroblasts is augmented during gestation in the presence of cortisol. The control and cortisol-augmented levels of FPF production, as determined by FPF ability to stimulate saturated phosphotidylcholine synthesis by lung epithelial Type II cells, is delayed during development in fibroblasts derived from male fetuses as compared to those derived from female fetuses. The mechanism by which this delay occurs has been addressed. Pregnant rats treated in vivo with dihydrotestosterone (DHT) showed decreased FPF activity from control or cortisol-treated fibroblasts derived from 20-day-old male or female fetuses. In vitro translated proteins of size-fractionated lung RNA from 19-day-old fibroblasts that were pretreated with DHT in vitro showed decreased FPF activity compared to nontreated samples. This decreased FPF activity was present even if the DHT-pretreated cells were stimulated with cortisol prior to RNA preparation. Using a mouse model of testicular feminization that contains no receptors for androgens showed no change in the cortisol augmented FPF activity when the fibroblasts were pretreated with DHT. These data taken together suggest that the delayed FPF production of male-derived lung fibroblasts is a physiologic process which requires androgen receptors, and the mechanism by which androgens inhibit FPF production appears to affect events occurring mainly at a pretranslational level.

• Morrison RS
• Fibroblast growth factors: potential neurotrophic agents in the central nervous system.
• J Neurosci Res. 1987; 17: 99-101

• Wice B, Milbrandt J, Glaser L
• Control of muscle differentiation in BC3H1 cells by fibroblast growth factor and vanadate.
• J Biol Chem. 1987; 262: 1810-7
• Display abstract

• We have examined the control of actin isoform synthesis by pituitary-derived fibroblast growth factor and serum in BC3H1 cells, a tumor-derived nonfusing muscle cell line. Under differentiating conditions in BC3H1 cells, the synthesis of beta- and gamma-actin ceases, and the rate of alpha-actin synthesis is increased concomitant with cessation of cell growth. Addition of fetal calf serum to differentiated cells reverses the process, whereas the addition of pituitary-derived fibroblast growth factor inhibits synthesis of alpha-actin but fails to induce the synthesis of beta- and gamma-actin. Analysis of RNA from differentiated BC3H1 cells after the addition of fetal calf serum indicated that the serum-induced increase in beta- and gamma-actin synthesis reflected an increase in their mRNA levels. In contrast, the repression of alpha-actin synthesis by fetal calf serum or fibroblast growth factor appears to reflect the translation efficiency of alpha-actin mRNA. Fibroblast growth factor is a competence factor for BC3H1 cells which allows them to progress from G0 4 h into the G1 phase of the cell cycle. In order to understand the nature of the intracellular signals responsible for the effect of fibroblast growth factor, we treated cells with vanadate, a known inhibitor of tyrosine-specific protein phosphatases. Vanadate fully mimics the action of fibroblast growth on actin synthesis and creatine phosphokinase synthesis and causes BC3H1 cells to exit the G0 portion of the cell cycle, as demonstrated by the induction of the c-fos proto-oncogene following addition of serum, vanadate, or bovine pituitary-derived fibroblast growth factor to these cells. We conclude that repression of alpha-actin synthesis and induction of the synthesis of beta- and gamma-actin are under independent control and that the induction of beta- and gamma-nonmuscle actin synthesis following serum addition is independent from movement into the cell cycle, and dependent on as yet unidentified serum components. The rate of synthesis of alpha-actin can be controlled by a defined mitogenic polypeptide fibroblast growth factor, which in short term experiments primarily affects the rate of translation of alpha-actin mRNA. The repression by fibroblast growth factor is most likely due to activation of a tyrosine specific protein kinase(s).

• Nishikawa K, Nagao Y, Yoshitake Y, Masuda Y
• [Role of growth factors in the autonomous growth of tumor cells]
• Gan To Kagaku Ryoho. 1987; 14: 1975-9
• Display abstract

• Typical characteristic of tumor cells is a decreased dependence on the growth factors for promotion of their growth. Growth factor independence and autonomous growth of tumor cells has been thought to be due to the ability of the cells to produce growth factors. According to an autocrine hypothesis, growth factor can be secreted by tumor cells and then binds to its receptor on the same cells to create an uncontrolled growth stimulation. Although there has been much circumstantial evidence to support this hypothesis, an autocrine mechanism is still uncertain. We purified DSF (DNA synthesis factor) from an extract of rat Rhodamine fibrosarcoma. This factor with strong affinity for heparin-Sepharose was a basic protein with molecular weight of 17,000. These properties were similar to those of basic FGF (fibroblast growth factor). We found that primary cells from the sarcoma could grow in serum-free medium supplemented with the extract of the same sarcoma tissue. The extract was fractionated to purify proteins which supported the growth of primary cells. DSF, transferrin and albumin which localized in the tissue, not due to contamination of blood, were purified. When they were added to the serum-free medium, the cells grew at a similar rate to that in medium supplemented with serum. This finding suggests that the growth of sarcoma cells is stimulated by their own growth factor and by some environmental factors.

• Shaked A, Sherris D, Mayer L
• Isolation of fibroblast proliferation factor: distinction from interleukin-1.
• Curr Surg. 1987; 44: 487-90

• Chamberlain CG, McAvoy JW
• Evidence that fibroblast growth factor promotes lens fibre differentiation.
• Curr Eye Res. 1987; 6: 1165-9
• Display abstract

• Lens epithelial cells from newborn rats undergo changes characteristic of fibre differentiation when cultured with rat neural retina or with a soluble mitogenic factor present in calf retina-conditioned medium. Mitogens have been isolated from retina in other laboratories, but have not previously been shown to promote fibre differentiation in mammalian lens. We prepared eye-derived growth factors I and II and alpha- and beta-retina-derived growth factors from bovine retinas. These factors all promoted lens fibre differentiation in our culture system, as assessed by morphological changes and the appearance of fibre cell-specific crystallins. There is now strong evidence that these retina-derived factors are identical to the acidic and basic forms of fibroblast growth factor (FGF), which is present in a variety of tissues. We found that acidic and basic FGF from rat brain also promoted lens fibre differentiation, suggesting that FGF is the factor from the retina responsible for inducing lens fibre differentiation.

• Jones MB, King RJ, Kuehl TJ
• Production of a growth factor(s) by fibroblasts from the lungs of ventilated premature baboons and oxygen-exposed adult hamsters.
• Am Rev Respir Dis. 1987; 135: 997-1001
• Display abstract

• Among the pathologic findings in infants succumbing to bronchopulmonary dysplasia are peribronchiolar and alveolar fibrosis. We examined the possibility that endogenously produced growth factors might contribute to these fibrotic changes through stimulation of fibroblast replication, with a resulting increase in collagen synthesis. We cultured fibroblasts from the lungs of prematurely delivered baboons and adult hamsters exposed to 100% oxygen and tested the media from these cells to see if they produced a factor(s) that was mitogenic to normal fibroblasts. We found that lung fibroblasts from baboons delivered prematurely at 140 days of gestation and ventilated for 6 days with 100% oxygen produced a factor(s) that supported fibroblast growth. A similar factor was also secreted by fibroblasts from normal term newborn baboon lungs and from the lungs of adult hamsters exposed to 100% oxygen for 4 to 8 days. Growth factor was not secreted by lung fibroblasts from 140-day non-breathing or 24-h-ventilated premature baboons, adult baboons, or hamsters not exposed oxygen. The growth-promoting substance is apparently a progression factor that is heat labile and inactivated by exposure to trypsin. These findings indicate that exposure of both the premature and adult lung to high concentrations of oxygen causes the elaboration of a growth factor that may induce fibroblast hyperplasia. This action may result in an increased production of connective tissue proteins and thereby contribute to the development of the fibrosis seen in bronchopulmonary dysplasia.

• Mascarelli F, Raulais D, Counis MF, Courtois Y
• Characterization of acidic and basic fibroblast growth factors in brain, retina and vitreous chick embryo.
• Biochem Biophys Res Commun. 1987; 146: 478-86
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• We have purified acidic and basic fibroblast growth factors (c-aFGF, c-bFGF) from 11 day-old chick embryo brain, retina and vitreous by heparin-Sepharose chromatography and reverse phase HPLC. The analysis of their biological activity as well as their molecular weight indicates that they were analogous to basic or acidic human and bovine FGF. The ratio of c-aFGF to c-bFGF activity depended of the tissue. In brain c-aFGF represented 66% of the total mitogenic activity retained on the heparin-sepharose column and c-bFGF 34% while retina contained 16% of c-aFGF and 84% of c-bFGF; vitreous 78% of c-aFGF and 22% of c-bFGF. Like human aFGF, Heparin stimulated purified c-aFGF mitogenic activity in the absence of serum but inhibited the activity of the retina acid soluble extract, in the presence of foetal calf serum (FCS). Thus, chick embryo and adult human acidic and basic FGF respectively share the same biochemical properties. Since there are no blood vessels in chick retina or vitreous, their presence in these tissues suggests that angiogenesis is not the only role of these growth factors.

• Austgulen R, Hammerstrom J, Nissen-Meyer J
• In vitro cultured human monocytes release fibroblast proliferation factor(s) different from interleukin 1.
• J Leukoc Biol. 1987; 42: 1-8
• Display abstract

• Human monocytes cultured in vitro released factors with growth stimulatory effect upon human dermal fibroblasts. Monocytes in RPMI 1640 released a growth enhancing activity in nearly constant amounts during in vitro differentiation to macrophage-like cells. The growth stimulatory effect mediated by supernatants was reduced when lipopolysaccharide (LPS) or opsonized zymosan particles (OpZ) were added to monocytes during the first 5 days of in vitro culture, thereafter reaching similar levels. The release of interleukin 1 (IL-1) from monocytes was restricted to the first 2 days of culture. IL-1 production by unstimulated monocytes was negligible, while LPS induced a high release of IL-1 from monocytes. A rabbit antibody against human IL-1 abolished the IL-1 activity of the monocyte supernatants as assessed in a mouse thymocyte proliferation assay, but caused only a small reduction of the fibroblast proliferation activity. Thus, the fibroblast growth stimulatory activity mediated by monocytes was caused by factor(s) in addition to IL-1.

• Connolly DT, Stoddard BL, Harakas NK, Feder J
• Human fibroblast-derived growth factor is a mitogen and chemoattractant for endothelial cells.
• Biochem Biophys Res Commun. 1987; 144: 705-12
• Display abstract

• Human fibroblasts were found to produce a potent mitogen and chemoattractant for fetal bovine aortic endothelial cells. Homogenates from AG1523 and AG1518 foreskin, CCD18Lu lung, and CCD18Co colon fibroblasts produced half-maximal stimulation of endothelial cell growth at concentrations of 1-7 micrograms/ml. The factor was purified from large-scale cultures of the CCD18Co fibroblasts using cation exchange chromatography and heparin-Sepharose chromatography. Such preparations were mitogenic for endothelial cells in vitro at concentrations of about 5-10 ng/ml, and promoted chemotaxis at 0.1-1 ng/ml. Heparinase treatment of the cells prevented the chemotactic response. These properties suggest that the factor may be related to fibroblast growth factor.

• Gospodarowicz D, Ferrara N, Schweigerer L, Neufeld G
• Structural characterization and biological functions of fibroblast growth factor.
• Endocr Rev. 1987; 8: 95-114

• Schweigerer L, Neufeld G, Gospodarowicz D
• Basic fibroblast growth factor as a growth inhibitor for cultured human tumor cells.
• J Clin Invest. 1987; 80: 1516-20
• Display abstract

• Basic fibroblast growth factor (bFGF) stimulates the proliferation of many cells and it is found in a wide variety of normal or transformed tissues. As demonstrated here, bFGF is also present in cultured human Ewing's sarcoma cells. Unexpectedly, however, bFGF isolated from these cells inhibits their own proliferation, indicating that bFGF can act as an endogenous (autocrine) growth inhibitor for cultured Ewing's sarcoma cells. Since bFGF also inhibits the proliferation of some further tumor cells, but stimulates that of others, it can be considered a bifunctional regulator of tumor cell proliferation. The autocrine growth-inhibitory effect of bFGF in Ewing's sarcoma cells may explain the low mitotic activity of Ewing's sarcomas.

• Kitagawa K, Nishino H, Ogiso Y, Iwashima A
• Inhibition by pertussis toxin of fibroblast growth factor-stimulated hexose transport in Swiss 3T3 cells.
• Biochim Biophys Acta. 1987; 931: 110-3
• Display abstract

• Addition of fibroblast growth factor to quiescent cultures of Swiss 3T3 cells stimulated the membrane transport of 2-deoxyglucose. Treatment of the cells with pertussis toxin (islet-activating protein) inhibited fibroblast growth factor-stimulated hexose transport. 5'-Guanylyl imidodiphosphate (p[NH]ppG), a non hydrolyzable analogue of GTP, increased the number of hexose carriers in the plasma membrane of saponin-permeabilized cells. These results suggest that guanine nucleotide binding protein may be involved in the regulation of hexose transport system by fibroblast growth factor in Swiss 3T3 cells.

• Yoshida T et al.
• Genomic sequence of hst, a transforming gene encoding a protein homologous to fibroblast growth factors and the int-2-encoded protein.
• Proc Natl Acad Sci U S A. 1987; 84: 7305-9
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• hst is a transforming gene first identified from transformed NIH 3T3 cells that were transfected with DNA of a human stomach cancer. A genomic fragment of hst obtained directly from a human genomic library also has transforming activity. This fragment has a coding sequence identical to that of the hst cDNA prepared from an NIH 3T3 transformant induced by DNA from a stomach cancer. The deduced amino acid sequence of the hst protein is 43%, 38%, and 40% homologous, respectively, to human basic fibroblast growth factor, human acidic fibroblast growth factor, and mouse int-2 protein in selected regions. This suggests that hst encodes a protein related to fibroblast growth factors, which are wide-spectrum mitogens, and to the int-2 protein, a potential oncogene product implicated in murine mammary carcinogenesis.

• Schweigerer L, Neufeld G, Mergia A, Abraham JA, Fiddes JC, Gospodarowicz D
• Basic fibroblast growth factor in human rhabdomyosarcoma cells: implications for the proliferation and neovascularization of myoblast-derived tumors.
• Proc Natl Acad Sci U S A. 1987; 84: 842-6
• Display abstract

• Cultured human embryonal rhabdomyosarcoma cells express the basic fibroblast growth factor (bFGF) gene and they produce bFGF, which is apparently composed of two microheterogenous forms with Mrs of 16,500 and 17,200, respectively. bFGF derived from the rhabdomyosarcoma cells stimulates their own proliferation and that of human or bovine vascular endothelial cells. It is conceivable that the rhabdomyosarcoma-derived bFGF stimulates the growth and neovascularization of human rhabdomyosarcomas and that it may thereby contribute to the development of these tumors.

• Marx JL
• Angiogenesis research comes of age.
• Science. 1987; 237: 23-4

• Phadke K
• Fibroblast growth factor enhances the interleukin-1-mediated chondrocytic protease release.
• Biochem Biophys Res Commun. 1987; 142: 448-53
• Display abstract

• The unfractionated macrophage-conditioned medium stimulates the chondrocytes to produce high levels of proteases. Purified IL-1 preparations exhibit significantly lower activity towards chondrocytes. This IL-1 mediated effect can be enhanced in presence of fibroblast growth factor, suggesting that other factors may collaborate with IL-1, in events leading to the cartilage destruction in vivo.

• Janet T, Miehe M, Pettmann B, Labourdette G, Sensenbrenner M
• Ultrastructural localization of fibroblast growth factor in neurons of rat brain.
• Neurosci Lett. 1987; 80: 153-7
• Display abstract

• The distribution of fibroblast growth factor (FGF) at the ultrastructural level in the brain of young (15- and 20-day-old) and adult (3-month-old) rats was investigated by immunocytochemistry. Strong staining was observed in most neurons of the cortex of young rat brain. In the same brain area of adult rat many neurons were also stained intensely, while others were negative. Neurons in the other parts of the brain and especially in the adult rat, were generally more weakly stained. The reaction product was located in the cytoplasm of the neuronal cell bodies and their processes. Astrocytes, oligodendrocytes, microglial cells, meningeal cells, choroid epithelial cells, ependymal cells and capillary endothelial cells showed no staining.

• Thomas KA
• Fibroblast growth factors.
• FASEB J. 1987; 1: 434-40
• Display abstract

• Fibroblast growth factors (FGFs) are heparin-binding protein mitogens that induce division of most cultured cells derived from embryonic mesoderm and neuroectoderm. Terminally differentiated neurons also respond in vitro by eliciting outgrowth of neurites. In vivo, FGFs have been shown to induce DNA synthesis, cell migration, blood vessel growth, and dermal wound closure. The protein and nucleic acid sequences for two different FGFs, denoted acidic and basic FGF, have been determined and recognized to be homologous. Additional genes recently have been identified that extend this protein family.

• Dickson C, Peters G
• Potential oncogene product related to growth factors.
• Nature. 1987; 326: 833-833

• Gospodarowicz D
• Isolation and characterization of acidic and basic fibroblast growth factor.
• Methods Enzymol. 1987; 147: 106-19

• Neufeld G, Ferrara N, Schweigerer L, Mitchell R, Gospodarowicz D
• Bovine granulosa cells produce basic fibroblast growth factor.
• Endocrinology. 1987; 121: 597-603
• Display abstract

• Cultured bovine granulosa cells express the gene encoding basic fibroblast growth factor (bFGF). The bFGF gene is transcribed into 7.0- and 3.7-kilobase mRNA transcripts which are apparently translated into 16,000 mol wt bFGF-like growth factor. The granulosa cell-derived bFGF is bioactive, i.e. it can stimulate the proliferation of capillary endothelial or granulosa cells. This mitogenic effect is prevented by specific neutralizing anti-bFGF antibodies. Our results indicate that bFGF derived from granulosa cells can act as both autocrine and paracrine growth factor, and they further suggest that the factor may be involved in the development of the rich vasculature of the theca interna of the follicle.

• Vlodavsky I et al.
• Endothelial cell-derived basic fibroblast growth factor: synthesis and deposition into subendothelial extracellular matrix.
• Proc Natl Acad Sci U S A. 1987; 84: 2292-6
• Display abstract

• Bovine aortic and corneal endothelial cells synthesize a growth factor that remains mostly cell-associated but can also be extracted from the subendothelial extracellular matrix (ECM) deposited by these cells. The endothelial cell-derived growth factors extracted from cell lysates and from the extracellular matrix appear to be structurally related to basic fibroblast growth factor by the criteria that they bind to heparin-Sepharose and are eluted at 1.4-1.6 M NaCl, have a molecular weight of about 18,400, cross-react with anti-basic fibroblast growth factor antibodies when analyzed by electrophoretic blotting and immunoprecipitation, and are potent mitogens for bovine aortic and capillary endothelial cells. It is suggested that endothelium can store growth factors capable of autocrine growth promotion in two ways: by sequestering growth factor within the cell and by incorporating it into the underlying extracellular matrix.

• Katsuoka K, Hein R, Krieg T, Schell H, Hornstein OP
• Trichilemmoid carcinoma-derived cells produce a growth factor for fibroblasts and reveal fibroblast chemotactic activity.
• Arch Dermatol Res. 1987; 279: 418-20

• Halaban R, Ghosh S, Baird A
• bFGF is the putative natural growth factor for human melanocytes.
• In Vitro Cell Dev Biol. 1987; 23: 47-52
• Display abstract

• Normal human melanocytes, unlike pigment cells from metastatic melanomas, do not survive in culture in routine, serum-supplemented media. The search for natural growth factors for melanocytes has shown that mitogenic activity is ubiquitous in several tissues and in melanomas. Of several known growth factors tested, basic fibroblast growth factor (bFGF) was the only one mitogenic for melanocytes but only in the presence of cyclic-adenosine-monophosphate (cAMP) stimulators. The mitogenic activity toward melanocytes in tissues and melanoma cell extracts had high affinity for heparin and antibodies to bFGF synthetic peptides. These results suggest that one of the growth factors for melanocytes might be bFGF or a bFGF-like polypeptide and that autocrine production of bFGF-like molecules by melanoma cells may contribute to the malignant phenotype of melanocytes. Because acidic FGF (aFGF) did not stimulate growth, the receptors for bFGF on melanocytes might be significantly different from those for a FGF.

• Gospodarowicz D
• Fibroblast growth factor: structural and biological properties.
• Int J Rad Appl Instrum B. 1987; 14: 421-34

• Neufeld G, Gospodarowicz D, Dodge L, Fujii DK
• Heparin modulation of the neurotropic effects of acidic and basic fibroblast growth factors and nerve growth factor on PC12 cells.
• J Cell Physiol. 1987; 131: 131-40
• Display abstract

• Nerve growth factor (NGF) and acidic or basic fibroblast growth factor (aFGF and bFGF, respectively) induce neurite outgrowth from the rat pheochromocytoma cell line, PC12. The neurites induced by these three factors are stable for up to a month in cell culture in the continued presence of any of the above growth factors. bFGF (ED50 = 30 pg/ml) is 800 fold more potent in stimulating neurite outgrowth than aFGF (ED50 = 25 ng/ml) and 260 fold more potent than NGF (ED50 = 8 ng/ml). While the neurotropic activities of aFGF and NGF are potentiated by heparin, that of bFGF is both partially inhibited or stimulated, depending upon the concentration of bFGF. Radioreceptor binding experiments show that aFGF and bFGF bind to a common binding site on the PC12 cell surface. Affinity labeling studies demonstrate a single receptor with an apparent molecular weight of 145,000 daltons, which corresponds to the high molecular weight receptor identified in BHK-21 cells. NGF does not appear to compete with aFGF or bFGF for binding to the receptor. Heparin blocked the binding of bFGF to the receptor but had only a small inhibitory effect on the binding of aFGF to the receptor. Thus, it appears that heparin inhibition of the neurotropic effects of bFGF occurs, at least in part, by impairing the interaction of bFGF with the receptor, while having little effect on that of aFGF. The stimulatory effects of heparin on the neurotropic activity of aFGF, bFGF, and NGF may occur through a site not associated with the respective cellular receptor for the growth factors.

• Gautschi P, Frater-Schroder M, Bohlen P
• Partial molecular characterization of endothelial cell mitogens from human brain: acidic and basic fibroblast growth factors.
• FEBS Lett. 1986; 204: 203-7
• Display abstract

• Two endothelial cell growth factors have been isolated from human brain extracts by a procedure involving heparin-Sepharose affinity chromatography and reverse-phase HPLC. On the basis of molecular size, amino acid composition and extended amino-terminal sequences, the two proteins are structurally closely related to bovine acidic and basic fibroblast growth factors (aFGF, bFGF). The N-terminal sequences of human and bovine aFGF differ in two positions while those for bFGF are indistinguishable. Human aFGF is microheterogenous: in addition to aFGF two N-terminally truncated forms lacking the first and the seven first amino acids, respectively, were characterized. Human and bovine mitogens possess similar mitogenic activities.

• Panet R, Amir I, Atlan H
• Fibroblast growth factor induces a transient net K+ influx carried by the bumetanide-sensitive transporter in quiescent BALB/c 3T3 fibroblasts.
• Biochim Biophys Acta. 1986; 859: 117-21
• Display abstract

• The bumetanide-sensitive transport system performed a net efflux of K+ in serum deprived quiescent cells. The addition of partially purified fibroblast growth factor (FGF) to G0/G1 phase 3T3 fibroblasts induced a transient net influx of K+, carried out by the bumetanide-sensitive transport system for 2-6 minutes. The stimulation of the bumetanide-sensitive K+ influx by FGF was followed by stimulation of the ouabain-sensitive K+ influx. In addition, both the bumetanide-sensitive and the ouabain-sensitive K+ influxes were found to be similarly stimulated when the G0/G1 3T3 cells were treated with insulin. These results suggest that growth factors such as FGF and insulin induce a change in the action of the bumetanide-sensitive transporter from performing net K+ efflux along its concentration gradient to an uphill transport pumping of K+ into the cell. We propose, therefore, that the bumetanide-sensitive transporter contributes to the increase in the intracellular K+ (and probable Na+) stimulated by growth factors such as FGF and insulin in early G1 phase of the cell cycle.

• Baird A et al.
• Molecular characterization of fibroblast growth factor: distribution and biological activities in various tissues.
• Recent Prog Horm Res. 1986; 42: 143-205

• Mergia A, Eddy R, Abraham JA, Fiddes JC, Shows TB
• The genes for basic and acidic fibroblast growth factors are on different human chromosomes.
• Biochem Biophys Res Commun. 1986; 138: 644-51
• Display abstract

• Basic and acidic fibroblast growth factor (FGF) are related both structurally and functionally. A bovine basic FGF cDNA and a human acidic FGF genomic fragment were used as hybridization probes in Southern blot analysis of DNAs isolated from a panel of 30 mouse-human cell hybrids. The gene encoding basic FGF was assigned to human chromosome 4, and the gene for acidic FGF to human chromosome 5. The two growth factors which are presumed to have a common evolutionary ancestor are therefore not linked. A HindIII restriction fragment length polymorphism was detected for human basic FGF.

• Pettmann B, Labourdette G, Weibel M, Sensenbrenner M
• The brain fibroblast growth factor (FGF) is localized in neurons.
• Neurosci Lett. 1986; 68: 175-80
• Display abstract

• The physiological role of fibroblast growth factor (FGF), like many other characterized growth factors, is poorly understood. Therefore, it would be useful to know in which cell type this growth factor is present. Using monoclonal and polyclonal antibodies against FGF, we have localized this molecule in rat brain. Interestingly, FGF is found exclusively in neuronal cells. Since FGF stimulates the proliferation and maturation of glial cells in vitro, its involvement in the interaction between neuronal and glial cells in vivo is likely. A role in the triggering of reactive gliosis can also be hypothesized.

• Moenner M, Chevallier B, Badet J, Barritault D
• Evidence and characterization of the receptor to eye-derived growth factor I, the retinal form of basic fibroblast growth factor, on bovine epithelial lens cells.
• Proc Natl Acad Sci U S A. 1986; 83: 5024-8
• Display abstract

• Eye-derived growth factor I (EDGF-I), the retinal form of the basic fibroblast growth factor, has been purified to homogeneity from bovine retina by heparin-Sepharose chromatography. The radioiodinated EDGF-I retained full mitogenic activity and was used to study the interaction of the growth factor with bovine epithelial lens cells. We showed that 125I-labeled EDGF-I bound in a saturable and reversible manner to a specific cellular receptor. Scatchard analysis of the equilibrium binding gave a Kd of 53 X 10(-12) M with approximately equal to 20,000 binding sites per cell. Crosslinking experiments using two homobifunctional reagents induced the formation of a specific major complex with a Mr of approximately equal to 145,000, as determined by NaDodSO4/PAGE, and independent of reducing conditions. These data establish the existence of a receptor for the basic growth factor derived from neural tissues and give an estimation of the size of this receptor at Mr 130,000.

• Huang JS, Huang SS, Kuo MD
• Bovine brain-derived growth factor. Purification and characterization of its interaction with responsive cells.
• J Biol Chem. 1986; 261: 11600-7
• Display abstract

• A large-scale purification procedure for brain-derived growth factors, without chromatography in 0.1% trifluoracetic acid, has been developed. Two related brain-derived growth factors have been purified to homogeneity from bovine brain using this procedure involving ammonium sulfate fractionation, followed by chromatography on CM-Sephadex C-50, sulfated Sephadex G-50, and heparin-Sepharose 4B. Brain-derived growth factor A (BDGF-A, molecular weight approximately 16,000) and brain-derived growth factor B (BDGF-B, molecular weight approximately 17,000) were eluted from heparin-Sepharose 4B at 1.2 and 1.6 M NaCl, respectively. Both BDGF-A and BDGF-B have a pI value of 5.7, have the same specific mitogenic activity, and react with mouse anti-BDGF-A antiserum. Both growth factors have a broad spectrum of mitogenic activity (vascular and aorta endothelial cells, chondrocytes, osteoblasts, epithelial cells, glial cells, fibroblasts, and smooth muscle cells), with a half-maximum effect at 10-20 pM. The binding of BDGF to bovine aorta endothelial cells and Swiss mouse 3T3 cells has been characterized. Binding of 125I-BDGF-A to these cells reached equilibrium in less than 15 min. Scatchard plot analysis of the binding of 125I-BDGF-A to endothelial cells and 3T3 cells showed a single class of high-affinity receptor with Kd values of 20 +/- 5 and 13 +/- 3 pM and receptor numbers/cell of 7,000 +/- 1,000 and 20,000 +/- 3,000, respectively. BDGF-B competed for 125I-BDGF-A binding in the same manner as unlabeled BDGF-A, suggesting that BDGF-A and BDGF-B bind to the same receptor. Basic pituitary fibroblast growth factor appeared to be a weak inhibitor, whereas platelet-derived growth factor and epidermal growth factor had no effect on 125I-BDGF-A binding. Protamine, histone, polylysine, and polyarginine are potent inhibitors of the mitogenic activity of BDGF-A and BDGF-B and of binding of 125I-BDGF-A to responsive cells. The half-time (t1/2) of internalization and degradation of cell surface-bound 125I-BDGF is 3 h.

• Uhlrich S, Lagente O, Choay J, Courtois Y, Lenfant M
• Structure activity relationship in heparin: stimulation of non-vascular cells by a synthetic heparin pentasaccharide in cooperation with human acidic fibroblast growth factors.
• Biochem Biophys Res Commun. 1986; 139: 728-32
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• Heparin enhances strongly the mitogenic properties of human acidic fibroblast growth factor (h-aFGF) on hamster fibroblast (CCL 39) or bovine lens epithelial cells (BEL). We report here that a synthetic heparin pentasaccharide with high affinity for antithrombin III has the same effect as heparin at about the same concentration. Thus a pentasaccharidic sequence may represent the shortest heparin structure which interacts with h-aFGF.

• Olwin BB, Hauschka SD
• Identification of the fibroblast growth factor receptor of Swiss 3T3 cells and mouse skeletal muscle myoblasts.
• Biochemistry. 1986; 25: 3487-92
• Display abstract

• Two distinct fibroblast growth factors (FGF) were purified to homogeneity from bovine brain on the basis of their ability to stimulate skeletal muscle myoblast proliferation. These growth factors are also mitogenic for Swiss 3T3 cells and appear to be closely related to or identical with previously isolated anionic and cationic fibroblast growth factors. The half-maximum concentrations (EC50) for stimulation of myoblast DNA synthesis by the anionic and cationic growth factors were 30pM and 1pM, respectively. In contrast, an EC50 of 45 pM was observed for stimulation of 3T3 cell DNA synthesis by both growth factors. Binding of 125I-labeled anionic FGF was saturable with apparent Kd values of 45 pM and 11 pM and approximately 60 000 and 2000 receptor sites per cell for 3T3 cells and MM14 murine myoblasts, respectively. Unlabeled anionic and cationic FGF equally displaced 125I-labeled anionic FGF from 3T3 cells while cationic FGF was more potent than anionic FGF for displacement from skeletal muscle myoblasts, demonstrating that a single receptor binds the two distinct growth factors. Binding was specific for these factors since platelet-derived growth factor, insulin, insulin-like growth factor 1, epidermal growth factor, and nerve growth factor were unable to displace bound 125I-labeled anionic FGF from Swiss 3T3 cells. Chemical cross-linking of specifically bound 125I-labeled anionic FGF to 3T3 cells and MM14 myoblasts identified a single detergent-soluble FGF receptor with an apparent molecular weight of 165 000.

• Baird A, Mormede P, Bohlen P
• Immunoreactive fibroblast growth factor (FGF) in a transplantable chondrosarcoma: inhibition of tumor growth by antibodies to FGF.
• J Cell Biochem. 1986; 30: 79-85
• Display abstract

• Using a radioimmunoassay for bovine pituitary fibroblast growth factor (FGF), we have established the presence of the immunoreactive mitogen in extracts of a transplantable mouse chondrosarcoma. Both neutral and acidic extracts of the tumor contain an immunoreactive FGF (ir-FGF) that cross-reacts in a parallel and dose-dependent fashion in the radioimmunoassay. The ir-FGF is retained on heparin-Sepharose affinity columns and can be detected in the same molecular weight forms as rat pituitary FGF. Mice (C57/Bl) inoculated with the tumor (10 mg, im) show a decreased rate of tumor growth when passively immunized with the antiserum to FGF. The results establish the presence of FGF in this tumor and implicate its role in the etiology of its development.

• Gospodarowicz D, Neufeld G, Schweigerer L
• Fibroblast growth factor.
• Mol Cell Endocrinol. 1986; 46: 187-204

• Neufeld G, Gospodarowicz D
• Basic and acidic fibroblast growth factors interact with the same cell surface receptors.
• J Biol Chem. 1986; 261: 5631-7
• Display abstract

• Despite quantitative differences, the activity of basic and acidic fibroblast growth factors (FGF) on a wide variety of normal diploid cells derived from neuroectoderm and mesoderm is intrinsically similar. This suggests that they bind to the same cell surface receptors. This was investigated using a baby hamster kidney cell line (BHK-21) as a model. BHK-21 cell membrane components that interact with basic and acidic FGF have been identified by covalent cross-linking to their respective 125I-labeled ligands. Under appropriate conditions, basic and acidic 125I-FGF were cross-linked, using disuccinimidyl suberate, to two receptor species with apparent molecular masses of 145,000 and 125,000 daltons, respectively. The labeling of those receptors is inhibited when either native basic or acidic FGF are present in excess during incubation of cells with either acidic or basic 125I-FGF. Competition of basic 125I-FGF with increasing concentrations of native acidic FGF results in a preferential decrease in the labeling of the 125,000-dalton species, whereas competition of acidic 125I-FGF with increasing concentrations of native basic FGF leads to a preferential decrease in the labeling of the 145,000-dalton species. The data suggest that qualitatively both mitogens interact with the same 145,000- and 125,000-dalton receptor species. The different affinities displayed by acidic and basic FGF toward their common receptor molecules could explain why acidic FGF, depending on the cell type considered, is 20-100-fold less potent than basic FGF.

• Gospodarowicz D, Neufeld G, Schweigerer L
• Molecular and biological characterization of fibroblast growth factor, an angiogenic factor which also controls the proliferation and differentiation of mesoderm and neuroectoderm derived cells.
• Cell Differ. 1986; 19: 1-17

• Senior RM, Huang SS, Griffin GL, Huang JS
• Brain-derived growth factor is a chemoattractant for fibroblasts and astroglial cells.
• Biochem Biophys Res Commun. 1986; 141: 67-72
• Display abstract

• Bovine brain-derived growth factor (BDGF), a 16-17 kDa protein with biochemical properties resembling brain-derived acidic fibroblast growth factor (acidic FGF) and endothelial cell growth factor, was found to have potent chemotactic activity for bovine ligament fibroblasts, human skin fibroblasts and rat astroglial cells, maximal at 100-200 pg/ml. The chemotactic activity was completely blocked by protamine sulfate (5 ug/ml), an inhibitor of receptor-binding and mitogenic activity of BDGF. BDGF did not stimulate migration of human monocytes. These results indicate that the effects of BDGF 'in vivo' might extend to mesenchymal cell recruitment.

• Smith BT, Floros J, Post M
• Hormonal/intercellular control of lung maturation.
• Prog Clin Biol Res. 1986; 226: 141-6

• Logan A, Logan SD
• Distribution of fibroblast growth factor in the central and peripheral nervous systems of various mammals.
• Neurosci Lett. 1986; 69: 162-5
• Display abstract

• Fibroblast growth factor (FGF) is a potent, mesodermal cell mitogen which has been identified throughout the central nervous system of various mammals and shown to display no regional concentration differences within the brain. An FGF-like mitogen of similar specific activity is also identified in the peripheral nervous system. Although not myelin-derived, these factors may arise from some component, of neuronal or glial origin, common to the central and peripheral nervous systems.

• Shimizu N
• [Protein and polypeptide growth factors--fibroblast growth factor]
• Nippon Rinsho. 1986; 44: 24-7

• Bravo R, Macdonald-Bravo H
• Effect of pH on the induction of competence and progression to the S-phase in mouse fibroblasts.
• FEBS Lett. 1986; 195: 309-12
• Display abstract

• The pH dependence of competence induction and progression to the S-phase in quiescent stimulated cells has been studied. The results show that: (i) induction of competence by fibroblast growth factor in these cells is relatively independent of the external pH between pH 5.6-7.6; (ii) progression of cells to the S-phase is highly sensitive to pH and shows a dramatic increase between pH 6.8-7.2. These observations suggest that the intracellular alkalinization triggered by growth factors is fundamental for progression but not for competence induction.

• Gautschi-Sova P, Muller T, Bohlen P
• Amino acid sequence of human acidic fibroblast growth factor.
• Biochem Biophys Res Commun. 1986; 140: 874-80
• Display abstract

• The complete amino acid sequence of human brain acidic fibroblast growth factor (aFGF) has been established. Human aFGF consists of 140 amino acids and is highly homologous to bovine aFGF (11 amino acid replacements). Results from experiments involving alkylation of cysteine residues are compatible with the possibilities that in aFGF all three cysteines exist as free sulfhydryls, or alternatively, that a disulfide bridge is present but cannot be identified due to disulfide scrambling caused by the SH group of the remaining cysteine. A potential glycosylation site Asn114-Gly115-Ser116 is present in aFGF but the mitogen does not bind to lectins suggesting that it may not be glycosylated.

• Gimenez-Gallego G, Conn G, Hatcher VB, Thomas KA
• Human brain-derived acidic and basic fibroblast growth factors: amino terminal sequences and specific mitogenic activities.
• Biochem Biophys Res Commun. 1986; 135: 541-8
• Display abstract

• Extended amino terminal sequence determinations, made on both acidic and basic fibroblast growth factors from human brain, showed extensive homology with each other and with their respective bovine counterparts. Both human growth factors in the presence of heparin have equivalent specific mitogenic activities on human umbilical vein endothelial cells in culture whereas in the absence of heparin, the acidic mitogen is less than 1% as active as the basic growth factor.

• Mormede P, Baird A, Pigeon P
• Immunoreactive fibroblast growth factor (FGF) in rat tissues: molecular weight forms and the effects of hypophysectomy.
• Biochem Biophys Res Commun. 1985; 128: 1108-13
• Display abstract

• Rat tissues were extracted at neutral pH with dilute (0.15 M) ammonium sulfate and examined for presence of fibroblast growth factor immunoreactivity (ir-FGF). Ir-FGF was found in substantial amounts in all the tissues but higher amounts were found in liver and kidney. Gel filtration column chromatography showed the presence of three molecular weight peaks of ir-FGF, one associated with the void volume (the major form in kidney extract), another with an apparent molecular weight of 150-180,000 Da (the major form in serum) and a third one of 70,000 Da (the major form in the other tissues examined: pituitary, brain, liver, spleen and thymus). No immunoreactivity coeluted with authentic FGF purified from bovine pituitary glands, suggesting that FGF is present in neutral extracts in association with binding molecules and/or precursors. Hypophysectomy had no major effect on ir-FGF in the tissues, but increased its amount in serum. The results establish a wide distribution of FGF in several embryologically, morphologically and developmentally distinct tissues and indicate that the pituitary is not the major source of FGF immunoreactivity, although it can be involved in the regulation of plasma levels.

• Smith BT, Torday JS
• Paracrine regulation of lung maturation.
• Pediatr Pulmonol. 1985; 1: 4952-4952
• Display abstract

• Although there is much yet to be learned about the regulation of fetal lung maturation, the study of this process has yielded much information that may serve as a paradigm for the understanding of fetal growth regulation in general. Two particularly interesting fetal adjustments of general regulatory phenomena have emerged: the ability of local tissue metabolism to anatomically or temporally isolate endocrine effects and the role of paracrine factors in coordinating the responses of different cells in complex organs.

• Weiss JB, Taylor CM, Wiseman D, Odedra R, Elstow S
• Angiogenic factor in ocular fluid.
• Lancet. 1985; 2: 1190-1

• Dodge WH
• Avian monocytic leukemia cells release fibroblast growth factor: implications to associated myelofibrosis.
• Leuk Res. 1985; 9: 1559-64
• Display abstract

• The marrow of chicks with leukemia induced by avian 'myeloblastosis' virus (AMV) exhibited a 5-10-fold increase in the number of fibroblast colony-forming cells (CFU-F). The increased CFU-F correlated with a mild fibrosis which can be seen in the marrow of these animals. Fibroblast proliferation likely was not simply due to the presence of leukemic cells because addition of formaldehyde-fixed peripheral leukemic cells failed to initiate CFU-F growth. Conditioned medium (CM) from day-4 cultures of peripheral leukemic cells was markedly stimulatory to CFU-F growth. The stimulatory activity was not due to virus released from the leukemic cells as, (1) removal of virus by pelleting had no effect on the CM activity and (2) direct inoculation of CFU-F cultures with virus failed to stimulate CFU-F growth. Normal avian marrow macrophage monolayers also released high levels of a fibroblast growth factor and both the macrophage-derived and leukemic cell-derived factors were heat-labile (65 degrees, 30 min).

• Askanas V, Gallez-Hawkins G
• Synergistic influence of polypeptide growth factors on cultured human muscle.
• Arch Neurol. 1985; 42: 749-52
• Display abstract

• In two- to five-week tissue cultures of biopsied adult human skeletal muscle, combined addition to the culture medium of insulin, fibroblast growth factor, and epidermal growth factor synergistically increased creatine kinase activity 17-fold, increased acetylcholine receptors tenfold, and accelerated muscle differentiation. This study provides the first demonstration of the beneficial influence of these peptides on human muscle. It also establishes a new culture medium, resulting in the following: (1) much better long-term growth and differentiation of biopsied adult human muscle; and (2) by allowing elimination of embryo extract and reduction of serum, an important step toward developing a fully defined medium for culturing biopsied adult human normal and pathologic muscle tissue.

• Cuny R, Zalik SE
• Comparison of newt lens regeneration stimulating activity in preparations of mammalian thyrotropin and fibroblast growth factor purified by various methods.
• Exp Eye Res. 1985; 41: 629-37
• Display abstract

• Irises of the newt Notophthalmus viridescens will regenerate a new lens in organ culture in the presence of the bovine thyrotropin preparation NIH-TSH-B8. It is not certain, however, whether thyrotropin itself is responsible for this stimulatory effect. To elucidate this problem further we compared the lens regeneration stimulating activity of thyrotropin preparations from several species, prepared by various methods. The lowest effective concentrations were approximately 3.0 micrograms ml-1 for the bovine NIH-TSH-B8 and 1.4 micrograms ml-1 for the ovine NIAMDD-oTSH-9 preparations. At those lowest concentrations, lenses with elongating lens fiber cells (stage 6) and enlarged lens fiber core (stage 8) were obtained, respectively, and the lens-fiber-specific protein gamma-crystallin was present in both cases. The crude bovine thyrotropin fraction, Sigma-TS-10, stimulated lens regeneration only at the highest concentration, 1400 micrograms ml-1. Bovine Pierce-bTSH, the purest thyrotropin preparation, stimulated lens regeneration sporadically at the lower concentration of 0.04 micrograms ml-1 up to the advanced stage 9 with large lens fiber core and flattened lens epithelium in one of nineteen irises. The pituitary fibroblast growth factor is a known contaminant of thyrotropin preparations. The preparation CR-FGF-40002 at concentrations between 0.001 and 0.1 microgram ml-1 did not promote lens regeneration. Therefore, the lens regeneration stimulating activity in thyrotropin preparations is not attributable to the fibroblast growth factor, and may also be independent of thyrotropin because the lens regeneration stimulating activity is not proportional to the thyrotropic activity in the preparations examined.

• Eccleston PA, Silberberg DH
• Fibroblast growth factor is a mitogen for oligodendrocytes in vitro.
• Brain Res. 1985; 353: 315-8
• Display abstract

• The effect of fibroblast growth factor (FGF) on oligodendrocyte development has been studied using dissociated mixed brain cells, cultured in a previously described serum-free medium. A greater number of galactocerebroside-positive oligodendrocytes could be demonstrated after 7 days in the presence of FGF than in control values. Using combined immunofluorescence and autoradiography an increased [3H]thymidine incorporation by galactocerebroside-positive oligodendrocytes was demonstrated after various times of exposure to FGF.

• Neufeld G, Gospodarowicz D
• The identification and partial characterization of the fibroblast growth factor receptor of baby hamster kidney cells.
• J Biol Chem. 1985; 260: 13860-8
• Display abstract

• The binding of biologically active, 125I-labeled basic fibroblast growth factor (FGF) to baby hamster kidney-derived cell line cells (BHK-21) was studied at 4 degrees C. Unlabeled FGF displaced cell surface bound 125I-FGF, but platelet-derived growth factor, epidermal growth factor, insulin, or transferrin did not. Binding was saturable both as a function of time and as a function of increasing 125I-FGF concentrations. Scatchard analysis of the binding data revealed the presence of about 1.2 X 10(5) binding sites/cell with an apparent KD of 270 pM. The number of the binding sites was down-regulated following preincubation of the cells with FGF. The density of binding sites/cell also decreased as an inverse function of cell density. When 125I-FGF binding was studied in a BHK-21 cell membrane preparation, it was found that the membranal binding site displayed a lower KD of 21 pM. 125I-FGF was covalently cross-linked to its cell surface receptor on intact BHK-21 cells using the homobifunctional agent disuccinimidyl suberate. Two macromolecular species with an apparent molecular weight of 145,000 and 125,000, respectively, were labeled under both reducing and nonreducing conditions. Unlabeled FGF competed with 125I-FGF for binding to both macromolecular species. The labeling of the macromolecules was also inhibited by heparin. No labeling was observed in the absence of the cross-linkers or when heat-inactivated 125I-FGF was used instead of radiolabeled, biologically active FGF.

• Klagsbrun M, Shing Y
• Heparin affinity of anionic and cationic capillary endothelial cell growth factors: analysis of hypothalamus-derived growth factors and fibroblast growth factors.
• Proc Natl Acad Sci U S A. 1985; 82: 805-9
• Display abstract

• Bovine hypothalamus-derived growth factors (HDGF), pituitary fibroblast growth factor (FGF), and brain FGF were analyzed by chromatography on immobilized heparin and tested for the ability to stimulate the proliferation of capillary endothelial (CE) cells. Two distinct CE cell growth factors were found in hypothalamus, one anionic (aHDGF; pI of about 5) and one cationic (cHDGF; pI of about 8). Both aHDGF and cHDGF adhered tightly to immobilized heparin. They were eluted with 0.9-1.1 M NaCl and 1.3-1.5 M NaCl, respectively. Pituitary and brain FGF were also found to bind to immobilized heparin and to stimulate CE cell proliferation. Pituitary FGF was eluted at 1.4-1.6 M NaCl. The elution profile of brain FGF showed that two peaks of CE cell growth factor activity were eluted from the heparin column, one at 1.0 M NaCl and a second at 1.4-1.6 M NaCl. The tight binding of all of these growth factors to heparin (particularly aHDGF, whose binding is unexpected because of its negative charge) is presented as evidence that CE cell growth factors all share an affinity for heparin.

• Togari A, Dickens G, Kuzuya H, Guroff G
• The effect of fibroblast growth factor on PC12 cells.
• J Neurosci. 1985; 5: 307-16
• Display abstract

• PC12 cells, which differentiate morphologically and biochemically into sympathetic neuron-like cells when treated with nerve growth factor, also respond to fibroblast growth factor. Some of the changes induced by fibroblast growth factor are similar to those seen after nerve growth factor treatment. Specifically, pituitary fibroblast growth factor causes the formation of processes initially comparable to those produced by nerve growth factor. However, in contrast to the outgrowth induced by nerve growth factor, which continues for several days, the outgrowth of processes induced by fibroblast growth factor ceases after about 3 days, even though fresh fibroblast growth factor is added. After about 6 days the processes induced by fibroblast growth factor have virtually disappeared. In this regard the processes induced by fibroblast growth factor are very similar to those induced by dibutyryl cyclic adenosine 3':5'-monophosphate (dBcAMP). The addition of nerve growth factor and fibroblast growth factor together appears to produce a synergistic effect on process formation, as does the simultaneous addition of nerve growth factor and dBcAMP. Cells pretreated (or primed) with nerve growth factor are able to regenerate processes much more rapidly in the presence of nerve growth factor than cells which have not been pretreated. When fibroblast growth factor is added to cells primed with nerve growth factor, more rapid regeneration of processes also occurs. The regeneration of neurites in response to either factor is blocked by the addition of an inhibitor of methylation. The process formation induced by fibroblast growth factor is preceded, as is the outgrowth in response to nerve growth factor treatment, by an induction of ornithine decarboxylase, a decrease in the phosphorylation of a specific cytoplasmic protein, and an increase in the phosphorylation of a specific non-histone nuclear protein. The effects of fibroblast growth factor and of nerve growth factor on ornithine decarboxylase are additive. Fibroblast growth factor does not cause an increase in the activity of acetylcholinesterase; nerve growth factor does. Fibroblast growth factor does not appear to be acting through the nerve growth factor receptor. The binding of iodinated nerve growth factor to PC12 cells is specific and is not inhibited by the presence of fibroblast growth factor. In addition, anti-nerve growth factor serum does not interfere with the action of fibroblast growth factor.(ABSTRACT TRUNCATED AT 400 WORDS)

• Logan A, Berry M, Thomas GH, Gregson NA, Logan SD
• Identification and partial purification of fibroblast growth factor from the brains of developing rats and leucodystrophic mutant mice.
• Neuroscience. 1985; 15: 1239-46
• Display abstract

• High titres of fibroblast growth factor activity (assessed by mitogenicity for Balb/c 3T3 fibroblast cells in vitro) have been extracted from the brains of foetal and neonatal rats long before myelinogenesis commences, from the brains of hypomyelinated, leucodystrophic, murine mutants and from normal adult rats. Partial purification, by ion-exchange chromatography and gel filtration, indicates that the brain fibroblast growth factor activity from these sources is associated with very similar basic protein fractions. These results, together with the observation that none of the samples of partially purified basic fibroblast growth factor elicits the experimental allergic encephalomyelitis response attributed to myelin basic protein in the rat, suggest that basic fibroblast growth factor is not a degradation product of myelin basic protein.

• Potter SR, Bienenstock J, Goldstein S, Buchanan WW
• Fibroblast growth factors in scleroderma.
• J Rheumatol. 1985; 12: 1129-35
• Display abstract

• Sera from patients with progressive systemic sclerosis (PSS), compared to normal sera, contained increased fibroblast growth promoting factor(s) (FGPF) for autologous and heterologous PSS skin fibroblasts and fibroblasts from normal donors. The relative stimulatory effect of FGPF was unaltered by dilution, mixing with normal serum, or media refeeding. The mean value of 42 PSS sera tested showed growth promoting activity significantly greater than the mean level of activity obtained with 42 sex matched normals (p less than 0.001). FGPF in sera from 25 patients with rheumatoid arthritis did not differ from normal controls. No correlation was seen between growth enhancement and patient age or medications. FGPF was present in serum plasma and is unlikely from platelets because serotonin and purified platelet factor produced no equivalent growth stimulation. FGPF was nondialysable, heat labile and insensitive to ribonuclease A. Disulphide bond integrity was required since 20 mM dithiothreitol destroyed activity. FGPF was excluded by an XM-50 filter after Sephadex G100 gel filtration.

• Gospodarowicz D, Cheng J, Lui GM, Fujii DK, Baird A, Bohlen P
• Fibroblast growth factor in the human placenta.
• Biochem Biophys Res Commun. 1985; 128: 554-62
• Display abstract

• Fibroblast growth factor (FGF) has been purified 333,000-fold from human placenta by a combination of salt precipitation, cation-exchange chromatography, and Heparin-Sepharose affinity chromatography. Molecular weight (15-16 kDaltons), amino acid composition, bioactivity and immunological crossreactivity with bovine pituitary FGF indicate that the mitogens from the two species are closely related molecules.

• Ishikawa O, Ishikawa H
• Neurofibroma-derived fibroblast-stimulating factor in the serum of patients with neurofibromatosis (von Recklinghausen's disease).
• Arch Dermatol Res. 1985; 277: 433-8
• Display abstract

• The effect of serum from patients with von Recklinghausen's disease [neurofibromatosis (NF)] on the cell growth and DNA synthesis of cultured neurofibroma-derived cells (NF fibroblasts grown from explant cultures of cutaneous neurofibromas from seven NF patients, and Schwann cells from a single-cell-suspension culture of the sample from one of these patients) was examined. Serum from patients with NF, whether autologous or allogeneic, enhanced the growth and 3H-thymidine uptake of NF fibroblasts twice as much as non-NF serum. Further study revealed that a dialysable, low-molecular-weight factor isolated from pooled NF serum was responsible for the stimulation of NF fibroblasts. This factor did not influence the DNA synthesis of either dermal fibroblasts cultured from non-NF patients or of Schwann cells. In contrast, no dialysable fraction of pooled serum from healthy persons exhibited such an NF-fibroblast-stimulating activity.

• Baird A, Esch F, Gospodarowicz D, Guillemin R
• Retina- and eye-derived endothelial cell growth factors: partial molecular characterization and identity with acidic and basic fibroblast growth factors.
• Biochemistry. 1985; 24: 7855-60
• Display abstract

• Two retina-derived growth factors have been isolated on the basis of their ability to stimulate the proliferation of capillary endothelial cells in vitro. Gas-phase sequence analysis identified the amino-terminal sequence of the major form of the mitogen as being identical with residues 1-35 of bovine basic fibroblast growth factor (FGF). Amino-terminal sequence analysis of the second form identified 28 residues that are indistinguishable from those of brain acidic FGF (residues 1-28). The possibility that these retina-derived endothelial cell growth factors are related to, if not identical with, basic and acidic FGF is supported by observations that they have similar molecular weights (15000-16000), similar retention behavior on all steps of chromatography (ion-exchange, heparin-Sepharose), and similar amino acid compositions and that they cross-react with antibodies to basic and acidic FGF. The eye-derived growth factors, like FGF, are potent stimulators of capillary endothelial cell growth in vitro. The results identify the major retina-derived endothelial cell growth factor as indistinguishable from basic FGF and demonstrate the presence of an acidic FGF in the eye. They suggest that at least some of the mitogenic, angiogenic, and neovascularizing activities described as being present in the retina are due to the existence of FGF in this tissue. The implications of this finding on the etiology and pathophysiology of vasoproliferative diseases of the eye are discussed.

• Askanas V, Cave S, Gallez-Hawkins G, Engel WK
• Fibroblast growth factor, epidermal growth factor and insulin exert a neuronal-like influence on acetylcholine receptors in aneurally cultured human muscle.
• Neurosci Lett. 1985; 61: 213-9
• Display abstract

• Fibroblast growth factor (FGF), epidermal growth factor (EGF) and insulin added in combination to the culture medium in which normal human muscle was cultured caused a 4.0-fold (P less than 0.005) increase of the total number of nicotinic acetylcholine receptors (AChRs) and a 4.5-fold (P less than 0.001) increase in AChR aggregation. Individually, only FGF caused a 3.0-fold increase (P less than 0.005) in AChR aggregation, without influencing the total number of AChRs. To the contrary, insulin alone caused a 2.0-fold increase (P less than 0.05) in the total number of AChRs without influencing AChR aggregation. These findings show that these three polypeptide growth factors exert a neuronal-like influence on cultured human muscle in regard to AChRs.

• Dohlman JG, Cooke MP, Payan DG, Goetzl EJ
• Structural diversity of the fibroblast-activating factors generated by human blood monocytes and U937 cells.
• J Immunol. 1985; 134: 3185-92
• Display abstract

• The incubation of purified human blood monocytes with phytohemagglutinin (PHA) and of cultured U937 human monocyte-like cells with phorbol myristate acetate (PMA) evoked the generation of fibroblast-activating activity, as assessed by stimulation of the uptake of [3H]thymidine by human dermal fibroblasts. Filtration of the supernates from monocytes and U937 cells on Sephadex G-75 resolved fibroblast-activating factors of m.w. 25,000 to 40,000, designated FAF-M and FAF-U937, respectively, from smaller factors of an apparent m.w. of approximately 10,000. FAF-M and FAF-U937 were acidic by isoelectric focusing with respective pI values of 4.0 to 5.2 and 5.4 to 5.6. The smaller factors from both sources filtered on Sephadex G-25 in phosphate-buffered saline with an apparent m.w. of 10,000. However, filtration of the same factors on Sephadex G-25 in 0.1 M acetic acid revealed one predominant fibroblast-activating activity for each cell source of an apparent m.w. of 500 to 1000. The 500 to 1000 dalton factors were inactivated by treatment with trypsin and subtilisin, suggesting that the activity was attributable to fibroblast-activating peptides, termed FAP-M and FAP-U937. FAP-M and FAP-U937 each appeared to be composed of a predominant hydrophilic activity by reverse-phase high-performance liquid chromatography. Human blood monocytes and U937 monocytes both produce structurally diverse fibroblast-activating proteins and peptides, which may contribute to the immunologic regulation of wound healing and fibrosis.

• Warden CH, Friedkin M
• Regulation of phosphatidylcholine biosynthesis by mitogenic growth factors.
• Biochim Biophys Acta. 1984; 792: 270-80
• Display abstract

• Phosphatidylcholine (PC) biosynthesis in cultured 3T3 fibroblasts was increased in varying degrees by these mitogenic growth factors: fetal bovine serum, insulin, 12-O-tetradecanoylphorbol-13-acetate, epidermal growth factor, vasopressin, fibroblast growth factor and insulin-like growth factors I and II. PC synthesis was increased 2-4-fold by 10% serum, up to 4-fold by growth factors alone, and up to 8-fold by combinations of two or more growth factors. Single growth factors had no effect on the incorporation of [3H]choline into the acid-soluble precursors of PC, while serum or combinations of two or more mitogens could increase the incorporation of [3H]choline into acid-soluble material by up to 2-fold. Serum was shown to increase choline phosphorylation, choline kinase activity and the size of the phosphocholine pool. These data were utilized to calculate the radioactive specific activity of phosphocholine. Serum did not increase phosphocholine specific activity above control values; thus the increased incorporation of labelled choline into PC after serum stimulation resulted from increased PC synthesis and not from a simple change in specific activity of precursor phosphocholine.

• Nishioka K, Kobayashi Y, Katayama I, Sano S
• Reduced response of scleroderma fibroblasts to fibroblast growth factor.
• Arch Dermatol Res. 1984; 276: 224-8
• Display abstract

• Reactivity of scleroderma fibroblasts to lymphoid cell-derived fibroblast growth factor (FGF) was assessed in this study. The fibroblasts from the sclerotic lesion failed to respond to FGF, whereas those from scleroedematous lesions responded equally to normal fibroblast. Response of the fibroblast from sclerotic lesion was also lower than that of the fibroblast from mature scar. Fibroblasts obtained from three different layers of healthy skin, papillary dermis, reticular dermis, and reticular-subcutaneous layer, responded equally to FGF, whereas the fibroblast of reticular dermis from sclerotic lesion failed to respond to FGF. It is suggested that the fibroblast of reticular dermis in scleroderma is variously activated by some unknown factors, so that they do not have enough reserve to respond to further stimuli.

• Gospodarowicz D
• Biological activity in vivo and in vitro of pituitary and brain fibroblast growth factor.
• Symp Fundam Cancer Res. 1984; 37: 109-34

• Thomas KA, Rios-Candelore M, Fitzpatrick S
• Purification and characterization of acidic fibroblast growth factor from bovine brain.
• Proc Natl Acad Sci U S A. 1984; 81: 357-61
• Display abstract

• Acidic brain fibroblast growth factor has been purified a minimum of 35,000-fold to apparent homogeneity by a combination of differential salt precipitation, ion exchange chromatography, gel filtration, isoelectric focusing, and hydrophobic chromatography on a C4 reversed-phase HPLC column. Two microheterogeneous forms of the molecule are obtained with apparent molecular masses of 16,600 and 16,800 daltons. The mitogen is highly active with half-maximal stimulation of BALB/c 3T3 fibroblasts at about 40 pg/ml in an assay using incorporation of [methyl-3H]thymidine into DNA.

• Dohlman JG, Payan DG, Goetzl EJ
• Generation of a unique fibroblast-activating factor by human monocytes.
• Immunology. 1984; 52: 577-84
• Display abstract

• Purified human monocytes incubated with phytohemagglutinin (PHA), bacterial lipopolysaccharide (LPS), or serum-opsonized zymosan particles (OZ) generate human dermal fibroblast-activating activity, as assessed by increased fibroblast incorporation of [3H]-thymidine. A maximum concentration of fibroblast-activating activity was attained within 4 hr with OZ, whereas similar maximum levels required 12 hr with LPS and PHA. Sonicates of unstimulated monocytes had only minimal activity and the protein synthesis inhibitor cycloheximide suppressed significantly the appearance of fibroblast-activating activity, suggesting that the factors are generated prior to release. Filtration of supernates from OZ-stimulated monocytes on Sephadex G-75 yielded polydisperse fibroblast-activating activities, of which the major factors exhibited a mol. wt. of approximately 60,000 and 10,000. The supernates from PHA-stimulated monocytes had one predominant factor, termed fibroblast-activating factor of monocytes (FAF-M), with an apparent mol. wt. of 38,000 and a minor activity with a mol. wt. of 10,000. FAF-M was composed of two principles with isoelectric points of 5.1-5.2 and 4.0-4.2 and was free of interleukin-1, as determined by the absence of thymocyte-activating activity. FAF-M and other fibroblast-activating factors may contribute to wound healing and fibrosis in lesions characterized by mononuclear phagocyte infiltrates.

• Picco R, Durand L
• [Cultures of endothelial cells from human cornea--growth factors. Preliminary note]
• Bull Soc Ophtalmol Fr. 1984; 84: 181-5

• Mayer P, Lam C
• Porcine alveolar macrophages, isolation, morphological and functional characteristics.
• Zentralbl Veterinarmed A. 1984; 31: 59-71

• Katoh Y, Takayama S
• Characterization of the growth responses of hamster fibroblasts and chondrogenic cells to fibroblast growth factor.
• Exp Cell Res. 1984; 150: 131-40
• Display abstract

• The effects of fibroblast growth factor (FGF) on hamster dermal fibroblasts and chondrogenic cells, both of mesodermal origin, were compared with special reference to growth stimulation and morphological changes in monolayer cultures, and colony formation in semisolid medium. FGF (10 to 200 ng/ml) caused appreciable cell proliferation of dermal fibroblasts but not of chondrogenic cells, while FGF (50-200 ng/ml) caused very marked dose-dependent morphological changes in monolayer cultures and colony formation in semisolid medium of both fibroblasts and chondrogenic cells. It is suggested that FGF is the same type of growth factor as the transforming growth factor(s) because, like the latter, it induces drastic morphological changes of normal mesodermal cells in monolayer cultures and their colony formation in semisolid medium.

• Kobayashi Y
• [Effects of the supernatants of the peripheral mononuclear cells activated by Con A-Sepharose on fibroblasts derived from the patients with PSS]
• Nippon Hifuka Gakkai Zasshi. 1984; 94: 599-609

• Potter SR, Bienenstock J, Lee P, Wilkinson S, Buchanan WW
• Clinical associations of fibroblast growth promoting factor in scleroderma.
• J Rheumatol. 1984; 11: 43-7
• Display abstract

• Normal adult human dermal fibroblasts were cultured in the presence of sera from 142 subjects. Results of a fibroblast proliferation assay revealed that 16 of 42 patients with scleroderma, 1 of 25 with rheumatoid arthritis, 3 of 10 with systemic lupus erythematosus, 2 of 3 with mixed connective tissue disease and 0 of 42 normal controls had values outside the normal range. The activity of fibroblast growth promoting factor (FGPF) in the scleroderma group correlated with the skin involvement but not with involvement of any other organ system. The levels of FGPF were higher in the first 2 years of disease duration than at any other time. Our data suggest that fibroblast activation may be a key process in the pathogenesis of scleroderma.

• Kagawa S
• [In-vitro cancer cell growth regulation by fibroblasts, with special reference to the relationship with fibroblast aging]
• Nippon Hinyokika Gakkai Zasshi. 1984; 75: 1718-21

• Wahl SM
• Lymphocyte and macrophage derived fibroblast growth factors.
• Prog Clin Biol Res. 1984; 154: 143-54

• Gospodarowicz D, Cheng J, Lui GM, Baird A, Bohlent P
• Isolation of brain fibroblast growth factor by heparin-Sepharose affinity chromatography: identity with pituitary fibroblast growth factor.
• Proc Natl Acad Sci U S A. 1984; 81: 6963-7
• Display abstract

• Brain and pituitary fibroblast growth factors (FGF) have been purified to apparent homogeneity from crude tissue extracts by a three-step procedure, including salt precipitation, ion-exchange chromatography, and heparin-Sepharose affinity chromatography. Brain and pituitary FGF have similar amino acid compositions and are indistinguishable with respect to molecular weight (16,000 by polyacrylamide gel electrophoresis), retention behavior in reversed-phase high-performance liquid chromatography, and recognition by antibodies directed against the amino-terminal sequence of pituitary FGF. Brain FGF preparations purified by heparin-Sepharose contain, in addition to the major FGF molecular species, at least two additional forms of the growth factor, which appear to be very similar by all the above criteria, except for retention in high-performance liquid chromatography.

• Post M, Floros J, Smith BT
• Inhibition of lung maturation by monoclonal antibodies against fibroblast-pneumonocyte factor.
• Nature. 1984; 308: 284-6
• Display abstract

• Fetal lung maturation, especially the onset of surfactant formation by alveolar type II cells, seems to be regulated by endogenous fetal glucocorticoids. Recent studies suggest that glucocorticoids do not act directly on the type II cell but rather on the lung mesenchyme. In response to glucocorticoids, the mesenchyme produces and secretes a polypeptide, fibroblast-pneumonocyte factor, which in turn stimulates surfactant synthesis by the alveolar type II cell. We report here the generation of hybridomas secreting monoclonal antibodies to rat lung fibroblast-pneumonocyte factor. Two monoclonal antibodies studied in detail reduced the cortisol-stimulated synthesis of saturated phosphatidylcholine in organotypic cultures of fetal rat lung cells and blocked the stimulatory effect of fibroblast-pneumonocyte factor in type II cells from these cultures. When embryonic chicks were injected on day 15 of incubation with either monoclonal antibody, they showed on days 20 and 21 biochemical evidence of delayed lung maturation as compared with controls. These effects were organospecific. Our observations support a physiological role for fibroblast-pneumonocyte factor in prenatal lung maturation.

• Westall FC, Rubin R, Gospodarowicz D
• Brain-derived fibroblast growth factor: a study of its inactivation.
• Life Sci. 1983; 33: 2425-9
• Display abstract

• Brain fibroblast growth factor has been identified as a component of myelin basic protein. Its activity is destroyed when treated with a variety of solvents including dilute acid, organic solvents and solutions of guanidine hydrochloride. These conditions do not alter the encephalitogenic properties of myelin basic protein.

• Wahl SM, Gately CL
• Modulation of fibroblast growth by a lymphokine of human T cell continuous T cell line origin.
• J Immunol. 1983; 130: 1226-30
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• Human peripheral blood T lymphocytes activated by mitogens or specific antigen produce a mediator that stimulates proliferation in a quiescent population of dermal fibroblasts. This lymphocyte product, which may have implications in the fibrosis associated with chronic cell-mediated inflammatory lesions, is not elaborated by unstimulated lymphocytes. Characterization of the fibroblast mitogenic factor differentiated it from a monocyte-derived factor with similar activity. The T cell factor appears to be a protein with an apparent m.w. of 40,000 and an isoelectric point between 5.0 and 5.5. This fibroblast growth factor can be produced in the relative absence of monocytes, and further evidence that the factor is a T cell product stems from the ability of a human T cell line to generate such a factor. Comparison of the biochemical characteristics of the normal T cell product with those of the T cell line revealed them to be indistinguishable. Thus, the T cell line can be used as a source of fibroblast-activating factor (FAF) for more detailed analysis of its structure and function. Further characterization of this lymphokine may contribute to understanding the mechanisms that mediate fibrosis in normal tissue repair as well as in the pathophysiologic fibrotic response associated with certain chronic inflammatory diseases.

• Jennings SD, Ham RG
• Clonal growth of primary cultures of human hyaline chondrocytes in a defined medium.
• Cell Biol Int Rep. 1983; 7: 149-59
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• Rapid clonal growth of primary cultures of human costal chondrocytes in a defined medium has been achieved. The basal nutrient medium used for such growth is MCDB 104. It is prepared without linoleic acid and supplemented with 1 microgram/ml insulin, 100 ng/ml fibroblast growth factor, 1.0 x 10(-7) M dexamethasone, and 5 micrograms/ml mixed lipids, presented to the cells in the form of liposomes. The lipid supplement contains soybean lecithin, cholesterol, sphingomyelin, vitamin E, and vitamin E acetate. No expression of cartilage-like differentiation occurs in the defined medium. However, colonies grown for several days in the defined medium and then grown for an additional period of time in medium F12 supplemented with fetal bovine serum and chicken embryo extract synthesize large amounts of refractile matrix that is stained intensely by acidified alcian green, thus demonstrating that the cells growing in the defined medium are capable of expressing cartilage matrix in a permissive environment. Good clonal growth and expression of differentiation can also be obtained by inoculating primary cultures of human chondrocytes directly into the F12-serum-embryo extract medium.

• Berry M et al.
• Deposition of scar tissue in the central nervous system.
• Acta Neurochir Suppl (Wien). 1983; 32: 31-53
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• Standard parasagittal lesions were placed stereotactically in the cerebral hemispheres of neonatal and adult rats in order to compare scarring in the immature and mature animal. Lesions were examined by light and electron-microscopy and immunofluorescence to study the astrocyte reaction, collagen deposition, and the formation of the basement membrane of the glia limitans. Normal mature scarring characterized by the deposition of collagen, astrocyte end-feet alignment over a glia limitans, and the permanent presence of mesodermal cells (fibroblasts and macrophages) in the core of the lesion, does not occur in wounds before 8-10 days post-partum (dpp). Instead there is no deposition of collagen, and only a transitory astrocyte response occurs with the formation of an interrupted glia limitans. These latter features disappear with time so that the wound is ultimately obliterated by the growth of axons and dendrites through the lesion. Mature scarring is attained over 8-12 dpp when increasing amounts of collagen are deposited and a continuous permanent glia limitans is formed. The acquisition of the mature response to injury from 8-12 dpp may be correlated with the presence of increasing titres of a fibroblast growth factor (FGF), derived from autolytic digestion of injured brain tissue. We have investigated FGF activity using a 3 T 3 fibroblast tissue culture assay to detect mitogenic activity in brain extracts from rats lesioned at different ages and from leukodystrophic mice which have no myelin. Our results show that high titres of FGF are present in the developing brain long before myelination commences, and that normal levels of FGF are found in the brains of leukodystrophic mice which have no myelin. Scarring in brain lesions in these mutants is quite normal.

• Togari A, Baker D, Dickens G, Guroff G
• The neurite-promoting effect of fibroblast growth factor on PC12 cells.
• Biochem Biophys Res Commun. 1983; 114: 1189-93
• Display abstract

• Treatment of PC12 cells with fibroblast growth factor(s) from either brain or pituitary caused neurite outgrowth comparable to that produced by nerve growth factor. The neurite outgrowth was preceded by a substantial rise in the activity of ornithine decarboxylase.

• Gospodarowicz D, Cheng J, Lirette M
• Bovine brain and pituitary fibroblast growth factors: comparison of their abilities to support the proliferation of human and bovine vascular endothelial cells.
• J Cell Biol. 1983; 97: 1677-85
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• The mitogenic effects of brain and pituitary fibroblast growth factors (FGF) on vascular endothelial cells derived from either human umbilical vein or bovine aortic arch have been compared. Both brain and pituitary FGF are mitogenic for low density human umbilical endothelial (HUE) cell cultures maintained on either fibronectin- or laminin-coated dishes or on biomatrices produced by cultured cells such as bovine corneal endothelial cells or the teratocarcinoma cell line PF-HR-9. Pituitary FGF triggered the proliferation of HUE cells at concentrations as low as 0.25 ng/ml, with a half-maximal response at 0.55 ng/ml and optimal effect at 2.5 to 5 ng/ml. It was 50,000-fold more potent than commercial preparations of endothelial cell growth factor and 40 times more potent than commercial preparations of pituitary FGF. Similar results were observed when the effect of pituitary FGF was tested on low density cultures of adult bovine aortic endothelial cells. When the activity of brain and pituitary FGF on low density HUE cell cultures was compared, both mitogens were active. To confirm the presence in brain extract of both acidic and neutral, as well as of basic mitogen, for HUE cells, brain tissues were extracted at acidic (4.5), neutral (7.2), and basic (8.5) pH. The three types of extracts were equally potent in supporting the proliferation of either HUE or adult bovine aortic endothelial cells. When the various extracts were absorbed at pH 6.0 on a carboxymethyl Sephadex C-50 column, the neutral and basic extracts had an activity after adsorption similar to that of unadsorbed extracts. In contrast, extracts prepared at pH 4.5 lost 90-95% of their activity which was recovered in the adsorbed fraction containing FGF.

• Lemmon SK, Bradshaw RA
• Purification and partial characterization of bovine pituitary fibroblast growth factor.
• J Cell Biochem. 1983; 21: 195-208
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• A purification procedure and partial characterization of bovine pituitary fibroblast growth factor (FGF) are described. The steps of the published methods [3,4] which yield inhomogeneous material, were retained, with modifications. The final isolation, with an additional purification of approximately 20-fold, was achieved by electrophoresis in polyacrylamide gels at acid pH. The mitogenic peptide has a molecular weight of 14,500--15,000 as determined on SDS gels, chromatographs as a monomer in nondenaturing conditions, and is active at the picomolar level in effecting the incorporation of 3H-thymidine in Balb/c 3T3 cells. A preliminary amino acid composition is presented.

• Wahl SM
• Immunoregulation of fibroblast growth and function.
• Lymphokine Res. 1983; 2: 139-44

• Westall FC
• Cancer basic protein: a cellular growth factor in cancer cells?
• J Theor Biol. 1983; 103: 473-80
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• Cancer basic protein possesses amazingly similar, but not identical, properties to myelin basic protein. Because of these similarities a theory is presented that cancer basic protein is a cellular growth factor and is intricately involved in the abnormal growth processes occurring in cancer cells.

• Lemaire I, Rola-Pleszczynski M, Begin R
• Asbestos exposure enhances the release of fibroblast growth factor by sheep alveolar macrophages.
• J Reticuloendothel Soc. 1983; 33: 275-85
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• Interaction between free airway cells (FAC) and lung fibroblasts was studied in a sheep model of asbestosis. Three groups of six sheep each received, respectively, by repeated intratracheal instillations, saline (control), 328 mg (low dose), and 2282 mg (high dose) of UICC chrysotile B asbestos. Sixteen months after the first instillation, FAC obtained by segmental bronchoalveolar lavage (BAL) of sheep in each group were incubated for various intervals, and the effect of their culture supernatants (FAC-SN) on human embryonic lung fibroblast proliferation was determined. FAC-SN from control animals stimulated thymidine (3H-TdR) incorporation by lung fibroblasts two- to threefold compared to untreated cultures. Maximal stimulation was observed at 48 hr and was correlated with a significant increase of the fibroblast population at 72 hr. FAC population from control sheep consisted primarily of macrophages (79%) and lymphocytes (15%), and separation of these two cell populations indicated that only macrophages produced the fibroblast-stimulating activity. Production occurred within 1 hr of incubation and was maximal between 2 and 4 hr. This activity was nondialyzable and stable at 56 degrees C for 30 min, but was destroyed at 80 degrees C and low pH. Moreover, FAC-SN from sheep exposed to asbestos stimulated 3H-TdR incorporation by fibroblasts five- to sixfold compared to two- to threefold for control FAC-SN. This activity may modulate fibrogenesis and may be involved in the eventual fibrogenic response to asbestos.

• Otto AM, Jimenez de Asua L
• Microtubule-disrupting agents can independently affect the prereplicative period and the entry into S phase stimulated by prostaglandin F2 alpha and fibroblastic growth factor.
• J Cell Physiol. 1983; 115: 15-22

• Pawlikowski M, Jakubowski L, Kaluzewski B
• The antimitotic effect of bromocriptine on human fibroblasts.
• J Neural Transm. 1983; 56: 91-5
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• The effect of bromocriptine on tritiated thymidine incorporation into nuclei of human fibroblasts has been investigated. A dose--dependent decrease of the labelling index has been found. This observation suggests that the antimitotic action of bromocriptine is a more general phenomenon not restricted to the adenohypophysial cells.

• Watanabe R
• [The roles of cartilage-derived factor (CDF) and bone-derived growth factor (BDGF) on skeletal growth]
• Osaka Daigaku Shigaku Zasshi. 1982; 27: 186-201

• Bremer EG, Hakomori S
• GM3 ganglioside induces hamster fibroblast growth inhibition in chemically-defined medium: ganglioside may regulate growth factor receptor function.
• Biochem Biophys Res Commun. 1982; 106: 711-8

• Gambarini AG, Armelin HA
• Purification and partial characterization of an acidic fibroblast growth factor from bovine pituitary.
• J Biol Chem. 1982; 257: 9692-7
• Display abstract

• Isoelectric focusing has allowed us to fractionate pituitary extracts into basic (pI 8-9) and acidic (pI 4-5) fibroblast growth factor. The acidic fibroblast growth factor (a) is stable upon refocusing, (b) migrates as an acidic protein in urea-containing gel electrophoresis; (c) is not cell-specific, being active with fibroblasts, adrenal, and glial cells, and (d) is a heterogeneous protein fraction with active components of different pI values. The component of pI 4.7, purified to or near homogeneity by isoelectric focusing shows a single peak of activity (Mr = 12,000) in gel chromatography and a single protein band of apparent Mr = 15,000 in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Maximal restimulation of DNA synthesis initiation on serum-deprived 3T3 fibroblasts is achieved at 1-2 ng/ml; activity with rat glial cells (C6-3D) is less pronounced than with 3T3 fibroblasts.

• Wyler DJ, Rosenwasser LJ
• Fibroblast stimulation in schistosomiasis. II. Functional and biochemical characteristics of egg granuloma-derived fibroblast-stimulating factor.
• J Immunol. 1982; 129: 1706-10
• Display abstract

• The pathogenetic mechanisms that underlie hepatic fibrosis in schistosomiasis are unknown, but may be under the regulation of molecules secreted by the hepatic granulomas that encase the helminth eggs. Previous studies in mice demonstrated that isolated Schistosoma mansoni egg granulomas can elaborate in vitro substances that stimulate fibroblast proliferation and collagen synthesis. The present study provides the initial characterization of the granuloma-derived factor(s). Serum-free cell culture supernatants from isolated granulomas contained activity that stimulated the uptake of (3H)-thymidine by quiescent human dermal fibroblasts. This activity was present in fractions (Sephadex G-200 chromatography) with estimated m.w. of 30,000 to 40,000. Activity eluted with linear salt gradients from ion exchange columns (DEAE Sephadex) with 0.4 to 0.7 and 1.5 to 1.8 M NaCl (pH 8.4). Activity was present in fractions with approximate pI of 6 to 6.5 and 4, prepared by flat-bed isoelectrofocusing of crude supernatant in granular gel. Activity that stimulated thymocyte proliferation in an interleukin 1 (IL 1) assay was present in crude granuloma supernatants as well as in the partially purified fractions that contained fibroblast-stimulating activity. We conclude that the granuloma-derived fibroblast-stimulating factor has biologic activity similar to IL 1 but on the basis of m.w. and pI determination is distinct from IL 1.

• Tseng SC, Savion N, Stern R, Gospodarowicz D
• Fibroblast growth factor modulates synthesis of collagen in cultured vascular endothelial cells.
• Eur J Biochem. 1982; 122: 355-60
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• Vascular endothelial cells derived from adult bovine aortic arch can be grown in two ways, either in the presence or absence of fibroblast growth factor. The types of collagen produced by cultures under these two conditions have been compared. In the presence of fibroblast growth factor, cells grow in an orderly fashion, express their normal phenotype and synthesize primarily type III collagen plus collagens types IV and V at a ratio of 10:1:3. Cultures grown in the absence of the factor lose their orderly pattern of growth, lose polarity and normal phenotypic expression. They devote twice the proportion of total protein-synthesizing capacity to collagen, and now synthesize type I in addition to the other collagen types. The ratio of collagen types I:III:IV:V is approximately 30:70:1:13. The kinds of type V collagen chains expressed are also altered. Fibroblast growth factor appears to modulate collagen synthesis, the major component of the extracellular matrix, and indirectly modulates the phenotypic expression of cultured vascular endothelial cells. In atherosclerosis, type I collagen is found in association with the intimal layer. The disorderly growth and the abnormal production of type I collagen by these vascular endothelial cells cultured in the absence of fibroblast growth factor is a model for a number of pathological situations including atherosclerotic plaque formation.

• Omar RA, Lanks KW
• Enhanced transformation of mouse embryo cells by SV40 virus following treatment with fibroblast growth factor.
• Eur J Cancer Clin Oncol. 1982; 18: 605-6

• Westall FC, Seil FJ, Woodward WR, Gospodarowicz D
• Brain fibroblast growth factors do not stimulate myelination or remyelination in tissue culture.
• J Neurol Sci. 1981; 52: 239-43
• Display abstract

• Myelin basic protein fragments that stimulated proliferation of fibroblasts in vitro did not enchance myelin formation or remyelination in nervous system tissue cultures. Similar fragments are present during demyelination in multiple sclerosis, but the lack of myelin stimulating activity in vitro suggests that they may not play a role in remyelination in vivo.

• Carlone RL, Ganagarajah M, Rathbone MP
• Bovine pituitary fibroblast growth factor has neurotrophic activity on newt limb regenerates and skeletal muscles in vitro.
• Exp Cell Res. 1981; 132: 15-21

• Pruss RM, Bartlett PF, Gavrilovic J, Lisak RP, Rattray S
• Mitogens for glial cells: a comparison of the response of cultured astrocytes, oligodendrocytes and Schwann cells.
• Brain Res. 1981; 254: 19-35
• Display abstract

• We have identified two growth factors for cultured rat astrocytes: fibroblast growth factor, a peptide derived from either whole bovine brain, or pituitaries, and a growth factor in extracts of bovine pituitary which was previously identified as a Schwann cell mitogen. Oligodendrocytes in primary cultures derived from neonatal rat central nervous system divide only rarely if at all. These growth factors did not stimulate primary oligodendrocytes to divide. Occasionally cells found in suspension in long-term cultures of the central nervous system were enriched for cells which were identified as oligodendrocytes by the presence of galactocerebroside on their surface and myelin basic protein in their cytoplasm. When provided with a monolayer of irradiated 3T3 cells, these oligodendrocytes were able to spread out and extend elaborate branched processes typical of oligodendrocytes in the primary cultures. Unlike their counterparts in the primary cultures, these suspension-derived oligodendrocytes are capable of cell division as demonstrated by the uptake of [3H]thymidine and autoradiography.

• Neff NT, Bourret L, Miao P, Dice JF
• Degradation of proteins microinjected into IMR-90 human diploid fibroblasts.
• J Cell Biol. 1981; 91: 184-94
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• Erythrocyte ghosts loaded with 125I-labeled proteins were fused with confluent monolayers of IMR-90 fibroblasts using polyethylene glycol. Erythrocyte-mediated microinjection of 125I-proteins did not seriously perturb the metabolism of the recipient fibroblasts as assessed by measurements of rates of protein synthesis, rates of protein degradation, or rates of cellular growth after addition of fresh serum. A mixture of cytosolic proteins was degraded after microinjection according to expected characteristics established for catabolism of endogenous cytosolic proteins. Furthermore, withdrawal of serum, insulin, fibroblast growth factor, and dexamethasone from the culture medium increased the degradative rates of microinjected cytosolic proteins, and catabolism of long-lived proteins was preferentially enhanced with little or no effect on degradation of short-lived proteins. Six specific polypeptides were degraded after microinjection with markedly different half-lives ranging from 20 to 320 h. Degradative rates of certain purified proteins (but not others) were also increased in the absence of serum, insulin, fibroblast growth factor, and dexamethasone. The results suggest that erythrocyte-mediated microinjection is a valid approach for analysis of intracellular protein degradation. However, one potential limitation is that some microinjected proteins are structurally altered by the procedures required for labeling proteins to high specific radioactivities. Of the four purified proteins examined in this regard, only ribonuclease A consistently showed unaltered enzymatic activity and unaltered susceptibility to proteolytic attack in vitro after iodination.

• Gambarini AG, Armelin HA
• Pituitary fibroblast growth factors: partial purification and characterization.
• Braz J Med Biol Res. 1981; 14: 19-27
• Display abstract

• 1. Isoelectric focusing of fractions derived from pH 7.0 extracts of bovine pituitary glands has been used to separate acidic and basic fibroblast growth factor (FGD) activity. FGF activity was assayed by measuring the initiation of DNA synthesis in Swiss mouse 3T3 fibroblasts. 2. Acidic and basic FGF activity present in crude extracts were not separated by chromatography on CM-Sephadex presumably because of association between acidic FGF and basic proteins. 3. Isoelectric focusing was used to separate acidic (pl 4-5) and basic (pI 8-9)FGF. The activity of acidic FGF was stable in urea and beta-mercaptoethanol, whereas basic FGF was inactivated. The activity of acidic FGF precipitated at pH 4.5 and was recovered when the precipitate was solubilized at pH 8, whereas basic FGF was soluble and stable at pH 4.5. 4. Acidic FGF at 10 ng/ml was as active as 1% fetal calf serum for the initiation of DNA synthesis. This potency is comparable with homogeneous preparations of basic FGF described in the literature.

• Kellett JG, Tanaka T, Rowe JM, Shiu RP, Friesen HG
• The characterization of growth factor activity in human brain.
• J Biol Chem. 1981; 256: 54-8
• Display abstract

• The purification of fibroblast growth factor from bovine brain has been reported (Gospodarowicz, D., Bialecki, H., and Greenberg, G. (1978) J. Biol. Chem. 253, 3736-3743). Westall et al. (Westall, F. C., Lennon, V. A., and Gospodarowicz, D. (1978) Proc. Natl. Acad. Sci. U. S. A. 75, 4675-4678) showed that bovine brain fibroblast growth factor was composed of three fragments derived by limited proteolysis from myelin basic protein. In the present study using similar purification methods, we isolated a fraction enriched in growth factor activity from human brain. The mitogenic activity could not be resolved from myelin basic protein by chromatographic procedures but, upon isoelectric focusing, the mitogen and myelin basic protein were readily dissociated. At least two potent growth factors (pI values 7.2 to 7.4 and 8.1 to 8.6) were identified. Studies of a relatively crude basic extract of human brain suggested that the brain may contain a number of growth factors.

• Webber RJ, Fox RR, Sokoloff L
• In vitro culture of rabbit growth plate chondrocytes. 2. Chondrodystrophic mutants.
• Growth. 1981; 45: 269-78
• Display abstract

• The epiphyseal plates of newborn normal (Cd/?), dwarf (Dw/Dw) and achondroplastic (ac/ac) rabbits were grown in organ culture for 48 hours. The 3H-thymidine labeling index of proliferating zone chondrocytes from both mutants was below that of normals. The generation time of Dw/Dw animals was longer than that of normals (greater than 24 vs. 17 hours respectively). Inasmuch as the medium was supplemented with 10% fetal calf serum, which presumably contains somatomedin, these data suggest that the Dw/Dw mutant cannot be considered simply a model for pituitary dwarfism. Pituitary fibroblast growth factor (FGF), 50ng/ml, decreased the labeling index of chondrocytes from both Dw/Dw and ac/ac animals. Thus neither a lack of FGF nor an end-organ non-responsiveness to this factor seems to play a role in the pathogenesis of either growth disturbance.

• Roncari DA
• Hormonal influences on the replication and maturation of adipocyte precursors.
• Int J Obes. 1981; 5: 547-52
• Display abstract

• The establishment of a propagating cell culture system of adipocyte precursors is facilitating the elucidation of the regulation of fat cell development by hormones and growth factors. 17 beta-Estradiol significantly enhances the replication of cultured human omental adipocyte precursors. Androgens do not influence their growth. These findings suggest that estrogens account, at least partly, for the changes that occur in certain adipose tissue regions at puberty. While the physiological role of fibroblast growth factor is unknown, the bovine polypeptide increases the replicative rate of skin fibroblasts to a greater extent than that of adipocyte precursors. On the other hand, partially purified, novel bovine pituitary polypeptides promote the replication of human adipocyte precursors to an appreciably greater extent than fibroblast growth factor. We are further characterizing these putative 'adipocyte growth factors'. Porcine pancreatic polypeptide significantly inhibits the multiplication of human omental adipocyte precursors. These findings may account for the scarcity of pancreatic fat tissue, and suggest that pancreatic polypeptide may control the growth of adipose tissue through its inhibitory effect.

• Franco WP, Bracey AW, Franco KL, Hufnagel CA, Glogar DH, Kloner RA
• Fibroblastic growth factor and infarct size.
• Ann Intern Med. 1980; 93: 637-8

• Gospodarowicz D, Mescher AL
• Fibroblast growth factor and the control of vertebrate regeneration and repair.
• Ann N Y Acad Sci. 1980; 339: 151-74

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