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. 1991 Sep 2;114(6):1261–1273. doi: 10.1083/jcb.114.6.1261

Acidic fibroblast growth factor in the developing rat embryo

PMCID: PMC2289139  PMID: 1716635

Abstract

Compared to basic fibroblast growth factor (bFGF), a widely distributed, broad spectrum mitogen and mesoderm inducer, acidic fibroblast growth factor (aFGF) is reported to have an essentially neural distribution and to be undetectable in the early embryo. In the present investigation, we used immunoblotting and immunochemistry to assess the cellular and tissue distributions of aFGF and bFGF in 11-20- d rat embryos. Immunoblotting of crude and heparin-bound embryo extracts revealed faint bands at the expected 17-18-kD and predominant bands at an apparent molecular mass of 26 to 28-kD (despite reducing conditions) using multiple specific antibodies for aFGF and bFGF. Pretreatment with 8 M urea yielded 18-20-kD aFGF and bFGF and some 24- 26-kD bFGF. Immunoreactivity for both aFGF and bFGF was positive and similar in the cytoplasm, nuclei, and extracellular matrix of cells of neuroectodermal and mesodermal origin, while it was negative in endoderm-derived cells. The distribution of immunoreactive aFGF and bFGF also showed changes during development that were associated with the process of cellular and tissue differentiation. For example, intensity and extent of immunoreactivity for both peptides progressively increased in the middle layer of the spinal cord with increasing differentiation of the neural cells. The immunostaining patterns were very similar for aFGF and bFGF for each organ and at each stage. In conclusion, high molecular mass forms of immunoreactive aFGF and bFGF are present in the rat embryo. Acidic FGF and bFGF are both widely distributed in tissues of neuroectodermal and mesodermal origin, and their distribution was very similar.

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Selected References

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  1. Abraham J. A., Whang J. L., Tumolo A., Mergia A., Friedman J., Gospodarowicz D., Fiddes J. C. Human basic fibroblast growth factor: nucleotide sequence and genomic organization. EMBO J. 1986 Oct;5(10):2523–2528. doi: 10.1002/j.1460-2075.1986.tb04530.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson K. J., Dam D., Lee S., Cotman C. W. Basic fibroblast growth factor prevents death of lesioned cholinergic neurons in vivo. Nature. 1988 Mar 24;332(6162):360–361. doi: 10.1038/332360a0. [DOI] [PubMed] [Google Scholar]
  3. 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 Dec 31;24(27):7855–7860. doi: 10.1021/bi00348a001. [DOI] [PubMed] [Google Scholar]
  4. Baird A., Walicke P. A. Fibroblast growth factors. Br Med Bull. 1989 Apr;45(2):438–452. doi: 10.1093/oxfordjournals.bmb.a072333. [DOI] [PubMed] [Google Scholar]
  5. Burgess W. H., Maciag T. The heparin-binding (fibroblast) growth factor family of proteins. Annu Rev Biochem. 1989;58:575–606. doi: 10.1146/annurev.bi.58.070189.003043. [DOI] [PubMed] [Google Scholar]
  6. Canalis E., Lorenzo J., Burgess W. H., Maciag T. Effects of endothelial cell growth factor on bone remodelling in vitro. J Clin Invest. 1987 Jan;79(1):52–58. doi: 10.1172/JCI112807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Caruelle D., Groux-Muscatelli B., Gaudric A., Sestier C., Coscas G., Caruelle J. P., Barritault D. Immunological study of acidic fibroblast growth factor (aFGF) distribution in the eye. J Cell Biochem. 1989 Feb;39(2):117–128. doi: 10.1002/jcb.240390204. [DOI] [PubMed] [Google Scholar]
  8. Casscells W., Speir E., Sasse J., Klagsbrun M., Allen P., Lee M., Calvo B., Chiba M., Haggroth L., Folkman J. Isolation, characterization, and localization of heparin-binding growth factors in the heart. J Clin Invest. 1990 Feb;85(2):433–441. doi: 10.1172/JCI114456. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Clegg C. H., Linkhart T. A., Olwin B. B., Hauschka S. D. Growth factor control of skeletal muscle differentiation: commitment to terminal differentiation occurs in G1 phase and is repressed by fibroblast growth factor. J Cell Biol. 1987 Aug;105(2):949–956. doi: 10.1083/jcb.105.2.949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Davis J. B., Stroobant P. Platelet-derived growth factors and fibroblast growth factors are mitogens for rat Schwann cells. J Cell Biol. 1990 Apr;110(4):1353–1360. doi: 10.1083/jcb.110.4.1353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Delli Bovi P., Curatola A. M., Kern F. G., Greco A., Ittmann M., Basilico C. An oncogene isolated by transfection of Kaposi's sarcoma DNA encodes a growth factor that is a member of the FGF family. Cell. 1987 Aug 28;50(5):729–737. doi: 10.1016/0092-8674(87)90331-x. [DOI] [PubMed] [Google Scholar]
  12. Dickson C., Peters G. Potential oncogene product related to growth factors. 1987 Apr 30-May 6Nature. 326(6116):833–833. doi: 10.1038/326833a0. [DOI] [PubMed] [Google Scholar]
  13. Dreyer D., Lagrange A., Grothe C., Unsicker K. Basic fibroblast growth factor prevents ontogenetic neuron death in vivo. Neurosci Lett. 1989 Apr 24;99(1-2):35–38. doi: 10.1016/0304-3940(89)90260-7. [DOI] [PubMed] [Google Scholar]
  14. Emoto N., Gonzalez A. M., Walicke P. A., Wada E., Simmons D. M., Shimasaki S., Baird A. Basic fibroblast growth factor (FGF) in the central nervous system: identification of specific loci of basic FGF expression in the rat brain. Growth Factors. 1989;2(1):21–29. doi: 10.3109/08977198909069078. [DOI] [PubMed] [Google Scholar]
  15. Esch F., Ueno N., Baird A., Hill F., Denoroy L., Ling N., Gospodarowicz D., Guillemin R. Primary structure of bovine brain acidic fibroblast growth factor (FGF). Biochem Biophys Res Commun. 1985 Dec 17;133(2):554–562. doi: 10.1016/0006-291x(85)90942-8. [DOI] [PubMed] [Google Scholar]
  16. Ferrari G., Minozzi M. C., Toffano G., Leon A., Skaper S. D. Basic fibroblast growth factor promotes the survival and development of mesencephalic neurons in culture. Dev Biol. 1989 May;133(1):140–147. doi: 10.1016/0012-1606(89)90305-9. [DOI] [PubMed] [Google Scholar]
  17. Finch P. W., Rubin J. S., Miki T., Ron D., Aaronson S. A. Human KGF is FGF-related with properties of a paracrine effector of epithelial cell growth. Science. 1989 Aug 18;245(4919):752–755. doi: 10.1126/science.2475908. [DOI] [PubMed] [Google Scholar]
  18. Florkiewicz R. Z., Sommer A. Human basic fibroblast growth factor gene encodes four polypeptides: three initiate translation from non-AUG codons. Proc Natl Acad Sci U S A. 1989 Jun;86(11):3978–3981. doi: 10.1073/pnas.86.11.3978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Folkman J., Klagsbrun M. Angiogenic factors. Science. 1987 Jan 23;235(4787):442–447. doi: 10.1126/science.2432664. [DOI] [PubMed] [Google Scholar]
  20. Folkman J., Klagsbrun M., Sasse J., Wadzinski M., Ingber D., Vlodavsky I. A heparin-binding angiogenic protein--basic fibroblast growth factor--is stored within basement membrane. Am J Pathol. 1988 Feb;130(2):393–400. [PMC free article] [PubMed] [Google Scholar]
  21. Gautschi-Sova P., Jiang Z. P., Fràter-Schröder M., Böhlen P. Acidic fibroblast growth factor is present in nonneural tissue: isolation and chemical characterization from bovine kidney. Biochemistry. 1987 Sep 8;26(18):5844–5847. doi: 10.1021/bi00392a039. [DOI] [PubMed] [Google Scholar]
  22. Globus R. K., Patterson-Buckendahl P., Gospodarowicz D. Regulation of bovine bone cell proliferation by fibroblast growth factor and transforming growth factor beta. Endocrinology. 1988 Jul;123(1):98–105. doi: 10.1210/endo-123-1-98. [DOI] [PubMed] [Google Scholar]
  23. Gonzalez A. M., Buscaglia M., Ong M., Baird A. Distribution of basic fibroblast growth factor in the 18-day rat fetus: localization in the basement membranes of diverse tissues. J Cell Biol. 1990 Mar;110(3):753–765. doi: 10.1083/jcb.110.3.753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Gospodarowicz D. Fibroblast growth factor. Crit Rev Oncog. 1989;1(1):1–26. [PubMed] [Google Scholar]
  25. Hanneken A., Lutty G. A., McLeod D. S., Robey F., Harvey A. K., Hjelmeland L. M. Localization of basic fibroblast growth factor to the developing capillaries of the bovine retina. J Cell Physiol. 1989 Jan;138(1):115–120. doi: 10.1002/jcp.1041380116. [DOI] [PubMed] [Google Scholar]
  26. Hauschka P. V., Mavrakos A. E., Iafrati M. D., Doleman S. E., Klagsbrun M. Growth factors in bone matrix. Isolation of multiple types by affinity chromatography on heparin-Sepharose. J Biol Chem. 1986 Sep 25;261(27):12665–12674. [PubMed] [Google Scholar]
  27. Houck K. A., Zarnegar R., Muga S. J., Michalopoulos G. K. Acidic fibroblast growth factor (HBGF-1) stimulates DNA synthesis in primary rat hepatocyte cultures. J Cell Physiol. 1990 Apr;143(1):129–132. doi: 10.1002/jcp.1041430117. [DOI] [PubMed] [Google Scholar]
  28. Hébert J. M., Basilico C., Goldfarb M., Haub O., Martin G. R. Isolation of cDNAs encoding four mouse FGF family members and characterization of their expression patterns during embryogenesis. Dev Biol. 1990 Apr;138(2):454–463. doi: 10.1016/0012-1606(90)90211-z. [DOI] [PubMed] [Google Scholar]
  29. Iberg N., Rogelj S., Fanning P., Klagsbrun M. Purification of 18- and 22-kDa forms of basic fibroblast growth factor from rat cells transformed by the ras oncogene. J Biol Chem. 1989 Nov 25;264(33):19951–19955. [PubMed] [Google Scholar]
  30. Joseph-Silverstein J., Consigli S. A., Lyser K. M., Ver Pault C. Basic fibroblast growth factor in the chick embryo: immunolocalization to striated muscle cells and their precursors. J Cell Biol. 1989 Jun;108(6):2459–2466. doi: 10.1083/jcb.108.6.2459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Kimelman D., Abraham J. A., Haaparanta T., Palisi T. M., Kirschner M. W. The presence of fibroblast growth factor in the frog egg: its role as a natural mesoderm inducer. Science. 1988 Nov 18;242(4881):1053–1056. doi: 10.1126/science.3194757. [DOI] [PubMed] [Google Scholar]
  32. Kimelman D., Kirschner M. W. An antisense mRNA directs the covalent modification of the transcript encoding fibroblast growth factor in Xenopus oocytes. Cell. 1989 Nov 17;59(4):687–696. doi: 10.1016/0092-8674(89)90015-9. [DOI] [PubMed] [Google Scholar]
  33. Kimelman D., Kirschner M. Synergistic induction of mesoderm by FGF and TGF-beta and the identification of an mRNA coding for FGF in the early Xenopus embryo. Cell. 1987 Dec 4;51(5):869–877. doi: 10.1016/0092-8674(87)90110-3. [DOI] [PubMed] [Google Scholar]
  34. Lipton S. A., Wagner J. A., Madison R. D., D'Amore P. A. Acidic fibroblast growth factor enhances regeneration of processes by postnatal mammalian retinal ganglion cells in culture. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2388–2392. doi: 10.1073/pnas.85.7.2388. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Liu L. M., Nicoll C. S. Evidence for a role of basic fibroblast growth factor in rat embryonic growth and differentiation. Endocrinology. 1988 Oct;123(4):2027–2031. doi: 10.1210/endo-123-4-2027. [DOI] [PubMed] [Google Scholar]
  36. Marics I., Adelaide J., Raybaud F., Mattei M. G., Coulier F., Planche J., de Lapeyriere O., Birnbaum D. Characterization of the HST-related FGF.6 gene, a new member of the fibroblast growth factor gene family. Oncogene. 1989 Mar;4(3):335–340. [PubMed] [Google Scholar]
  37. Mascarelli F., Raulais D., Counis M. F., Courtois Y. Characterization of acidic and basic fibroblast growth factors in brain, retina and vitreous chick embryo. Biochem Biophys Res Commun. 1987 Jul 31;146(2):478–486. doi: 10.1016/0006-291x(87)90554-7. [DOI] [PubMed] [Google Scholar]
  38. Morrison R. S., Sharma A., de Vellis J., Bradshaw R. A. Basic fibroblast growth factor supports the survival of cerebral cortical neurons in primary culture. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7537–7541. doi: 10.1073/pnas.83.19.7537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Noji S., Matsuo T., Koyama E., Yamaai T., Nohno T., Matsuo N., Taniguchi S. Expression pattern of acidic and basic fibroblast growth factor genes in adult rat eyes. Biochem Biophys Res Commun. 1990 Apr 16;168(1):343–349. doi: 10.1016/0006-291x(90)91714-4. [DOI] [PubMed] [Google Scholar]
  40. Olwin B. B., Hauschka S. D. Fibroblast growth factor receptor levels decrease during chick embryogenesis. J Cell Biol. 1990 Feb;110(2):503–509. doi: 10.1083/jcb.110.2.503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Presta M., Statuto M., Rusnati M., Dell'Era P., Ragnotti G. Characterization of a Mr 25,000 basic fibroblast growth factor form in adult, regenerating, and fetal rat liver. Biochem Biophys Res Commun. 1989 Nov 15;164(3):1182–1189. doi: 10.1016/0006-291x(89)91794-4. [DOI] [PubMed] [Google Scholar]
  42. Renko M., Quarto N., Morimoto T., Rifkin D. B. Nuclear and cytoplasmic localization of different basic fibroblast growth factor species. J Cell Physiol. 1990 Jul;144(1):108–114. doi: 10.1002/jcp.1041440114. [DOI] [PubMed] [Google Scholar]
  43. Risau W., Ekblom P. Production of a heparin-binding angiogenesis factor by the embryonic kidney. J Cell Biol. 1986 Sep;103(3):1101–1107. doi: 10.1083/jcb.103.3.1101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Risau W., Gautschi-Sova P., Böhlen P. Endothelial cell growth factors in embryonic and adult chick brain are related to human acidic fibroblast growth factor. EMBO J. 1988 Apr;7(4):959–962. doi: 10.1002/j.1460-2075.1988.tb02901.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Sasse J., Sullivan R., Klagsbrun M. Purification of cartilage-derived growth factors. Methods Enzymol. 1987;146:320–325. doi: 10.1016/s0076-6879(87)46033-3. [DOI] [PubMed] [Google Scholar]
  46. Saxén L. Embryonic induction. Clin Obstet Gynecol. 1975 Dec;18(4):149–175. doi: 10.1097/00003081-197512000-00013. [DOI] [PubMed] [Google Scholar]
  47. Schweigerer L., Malerstein B., Neufeld G., Gospodarowicz D. Basic fibroblast growth factor is synthesized in cultured retinal pigment epithelial cells. Biochem Biophys Res Commun. 1987 Mar 30;143(3):934–940. doi: 10.1016/0006-291x(87)90340-8. [DOI] [PubMed] [Google Scholar]
  48. Seed J., Olwin B. B., Hauschka S. D. Fibroblast growth factor levels in the whole embryo and limb bud during chick development. Dev Biol. 1988 Jul;128(1):50–57. doi: 10.1016/0012-1606(88)90265-5. [DOI] [PubMed] [Google Scholar]
  49. Senior R. M., Huang S. S., Griffin G. L., Huang J. S. Brain-derived growth factor is a chemoattractant for fibroblasts and astroglial cells. Biochem Biophys Res Commun. 1986 Nov 26;141(1):67–72. doi: 10.1016/s0006-291x(86)80335-7. [DOI] [PubMed] [Google Scholar]
  50. Seno M., Iwane M., Sasada R., Moriya N., Kurokawa T., Igarashi K. Monoclonal antibodies against human basic fibroblast growth factor. Hybridoma. 1989 Apr;8(2):209–221. doi: 10.1089/hyb.1989.8.209. [DOI] [PubMed] [Google Scholar]
  51. Slack J. M., Darlington B. G., Heath J. K., Godsave S. F. Mesoderm induction in early Xenopus embryos by heparin-binding growth factors. Nature. 1987 Mar 12;326(6109):197–200. doi: 10.1038/326197a0. [DOI] [PubMed] [Google Scholar]
  52. Sommer A., Moscatelli D., Rifkin D. B. An amino-terminally extended and post-translationally modified form of a 25kD basic fibroblast growth factor. Biochem Biophys Res Commun. 1989 May 15;160(3):1267–1274. doi: 10.1016/s0006-291x(89)80140-8. [DOI] [PubMed] [Google Scholar]
  53. Taira M., Yoshida T., Miyagawa K., Sakamoto H., Terada M., Sugimura T. cDNA sequence of human transforming gene hst and identification of the coding sequence required for transforming activity. Proc Natl Acad Sci U S A. 1987 May;84(9):2980–2984. doi: 10.1073/pnas.84.9.2980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Thomas K. A., Rios-Candelore M., Giménez-Gallego G., DiSalvo J., Bennett C., Rodkey J., Fitzpatrick S. Pure brain-derived acidic fibroblast growth factor is a potent angiogenic vascular endothelial cell mitogen with sequence homology to interleukin 1. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6409–6413. doi: 10.1073/pnas.82.19.6409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Wagner J. A., D'Amore P. A. Neurite outgrowth induced by an endothelial cell mitogen isolated from retina. J Cell Biol. 1986 Oct;103(4):1363–1367. doi: 10.1083/jcb.103.4.1363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Whitman M., Melton D. A. Growth factors in early embryogenesis. Annu Rev Cell Biol. 1989;5:93–117. doi: 10.1146/annurev.cb.05.110189.000521. [DOI] [PubMed] [Google Scholar]
  57. Zhan X., Bates B., Hu X. G., Goldfarb M. The human FGF-5 oncogene encodes a novel protein related to fibroblast growth factors. Mol Cell Biol. 1988 Aug;8(8):3487–3495. doi: 10.1128/mcb.8.8.3487. [DOI] [PMC free article] [PubMed] [Google Scholar]

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