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Production of Growth Factors by the Blastema During Limb Regeneration of Urodeles (Amphibia)

  • Chapter
Recent Trends in Regeneration Research

Part of the book series: NATO ASI Series ((NSSA,volume 172))

Summary

The blastema of regenerating limbs of Amphibia contains neurotrophic factor(s) for spinal nerves and mitogenic factor(s) for blastema cells. The neurotrophic activity of blastemata was shown by co-culture of spinal cord fragments with blastemata by medium conditioned by blastemata or by blastema extracts; only the mesenchymal component of blastemata exhibited neurotrophic activity which appeared to be related to the proliferation rate of blastema cells. The mitogenic activity of blastemata was shown by using blastema extracts on cultured blastema cells; both the epidermal and mesenchymal components of blastemata were mitogenic for these cells. The nature of blastema growth factors has been approached by testing mitogenic activity of heparin on blastema cells, extraction of heparin-binding growth factors from blastemata and immunolocalization of aFGF. Our results show that both epidermal cap and blastema mesenchyme contain aFGF. This growth factor is probably involved in blastema cell proliferation during regeneration but not in the neurotrophic activity of blastemata.

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Abbreviations

aFGF:

acidic fibroblast growth factor

bFGF:

basic fibroblast growth factor

ECGF:

endothelial cell growth factor

EDGFII:

eye derived growth factor II

EGF:

epidermal growth factor

GGF:

glial growth factor

PDGF:

platelet derived growth factor

TGF:

transforming growth factor

HPLH:

high pressure liquid chromatography

GAG:

Glycosaminoglycan.

References

  • Albert, P., and Boilly, B., 1986, Evolution in vitro de cellules de blastèmes de régénération de membre d1Äxolotl: influence de l’insuline et d’extraits nerveux sur la prolifération cellulaire, Biol. Cell., 58:251–262.

    Article  PubMed  CAS  Google Scholar 

  • Albert, P., Boilly, B., Courty, J., and Barritault, D., 1987, Stimulation in cell culture of mesenchymal cells of newt limb blastemas by EDGFI or II (basic or acidic FGF), Cell. Diff., 21:63–68.

    Article  CAS  Google Scholar 

  • Baird, A., Esch, F., Mormede, P., Ueno, N., Ling, N., Bolhen, P., Ying, S.Y., Wehrenberg, W.B., and Guillemin, R., 1986, Molecular characterization of fibroblast growth factor: distribution and biological activities in various tissues, Recent Prog. Horm. Res., 42:177–205.

    Google Scholar 

  • Balk, S.D., Riley, T.M., Gunther, H.S., and Morisi, A., 1985, Heparin-treated, v-myc- transformed chicken heart mesenchymal cells assume a normal morphology but are hypersensitive to epidermal growth factor (EGF) and brain fibroblast growth factor (b FGF); cells transformed by the v-Ha-ras oncogene are refractory to FGF and b FGF but are hypersensitive to insulin-like growth factors, Proc. Natl. Acad. Sci. USA., 82:5781–5785.

    Article  PubMed  CAS  Google Scholar 

  • Bauduin, B., 1986, Influence neurotropedes blastèmes de regeneration du membre antérieur sur la moëlle épinière du triton, Pleurodeles waltlii. Etude in vitro. Thèse, Université de Lille.

    Google Scholar 

  • Bauduin, B., Lassalle, B., and Boilly, B., The effect of regenerating blastema on the cultured spinal cord of the newt, Pleurodeles waltlii. Differentiation (submitted) .

    Google Scholar 

  • Bell, K.M., 1986, The preliminary characterization of mitogens secreted by embryonic chick wing bud tissues in vitro, J. Embrvol. exp. Morph., 93:257–265.

    CAS  Google Scholar 

  • Boilly, B., and Albert, P., 1988a, Control of blastema cell proliferation during axolotl limb regeneration: in vitro cell culture study, in: Monogr. devl. Biol., 21:1–8, Karger, Basel.

    CAS  Google Scholar 

  • Boilly, B., and Albert, P., 1988b, Blastema cell proliferation in vitro: effects of limb amputation on the mitotic activity of spinal cord extracts, Biol. Cell., 62:183–188.

    PubMed  CAS  Google Scholar 

  • Boilly, B., and Bauduin, B., 1988, Production in vitro by spinal cord of growth factor(s) acting on newt limb regeneration: influence of regeneration of the nerve fibers, Dev. Brain Res., 38:155–160.

    Article  CAS  Google Scholar 

  • Boilly, B., Oudkhir, M., and Lassalle, B., 1985, Control of the blastemal cell cycle by the peripheral nervous system during newt limb regeneration: continuous labeling analysis, Biol. Cell., 55:107–112.

    Article  PubMed  CAS  Google Scholar 

  • Bottenstein, J.E., Skaper, S.D., Varon, S.S., and Sato, G.H., 1980, Selective survival of neurons from chick embryo sensory ganglion dissociated utilizing serum-free supplemented medium, Exp. Cell. Res., 125:183–190.

    Article  PubMed  CAS  Google Scholar 

  • Brockes, J.P., 1984, Mitogenic growth factors and nerve dependence of limb regeneration, Science, 255:1280–1287.

    Article  Google Scholar 

  • Brockes, J.P., and Kintner, C.B., 1986, Glial growth and nerve-dependent proliferation in the regeneration in the blastema of Urodele Amphibian, Cell., 45:301–306.

    Article  PubMed  CAS  Google Scholar 

  • Burnett, A.L., Kary, C.E., and Lagorio, A.M., 1971, Induction of growth in newt and frog limbs after perfusion with extracts from newt blastemas, Nature, 234:98–99.

    Article  PubMed  CAS  Google Scholar 

  • Carlone, R.L., and Mescher, A., 1985, Trophic factors from nerves; In: “Regulation of Vertebrate Limb Regeneration” (R.E. Sicard, ed.), pp. 93–105. Oxford Univ. Press.

    Google Scholar 

  • Castellot, Jr., J.J., Addonizio, M.L., Rosenberg, R., and Karnovsky, M.J., 1981, Cultured endothelial cells produce a heparin like inhibitor of smooth muscle cell growth, J. Cell. Biol., 90:372–379.

    Article  PubMed  CAS  Google Scholar 

  • Clowes, A.W., and Karnovsky, M.J., 1977, Suppression by heparin of smooth muscle cell proliferation in injured arteries, Nature, 265:625–626.

    Article  PubMed  CAS  Google Scholar 

  • Courty, J., Loret, C., Moenner, M., Chevallier, B., Lagente, O., Courtois, Y., and Barritault, D., 1985, Bovine retina contains three growth factors activities with different affinity to heparin: eye derived growth factor I, II, III, Biochimie, 67:265–269.

    Article  PubMed  CAS  Google Scholar 

  • Deck, J.D., and Futch, C.B., 1969, The effects of infused materials upon the regeneration of newt limbs: I. Blastemal extracts in denervated limb stumps, Develop. Biol., 20:332–348.

    Article  PubMed  CAS  Google Scholar 

  • Deck, J.D., and Dent, J.N., 1970, The effects of infused materials upon the regeneration of newt limbs: III Blastemal extracts and alkaline phosphatase in irradiated limb stumps, Anat. Rec., 168:525–536.

    Article  PubMed  CAS  Google Scholar 

  • Gautschi-Sova, P., Jiang, Z.P., Schroder, M., and Bohlen, P., 1987, Acidic fibroblast growth factor is present in non-neural tissue: isolation and chemical characterization from bovine kidney, Biochemistry, 26:5844–5847.

    Article  PubMed  CAS  Google Scholar 

  • Globus, M., Vethamany-Globus, S., and Lee, Y.C.I., 1980, Effect of apical epidermal cup on mitotic cycle and cartilage differentiation in regeneration blastema in the newt Notophthalmus viridescens. Develop. Biol., 75:358–372.

    Article  PubMed  CAS  Google Scholar 

  • Gospodarowicz, D., Manoglia, S., Cheng, J., and Fujii, D.K., 1986, Effect of fibroblast growth factor and lipoproteins on the proliferation of endothelial cells derived from bovine adrenal cortex, brain cortex and corpus luteum capillaries, J. Cell. Physiol., 127:121–136.

    Article  PubMed  CAS  Google Scholar 

  • Goss, R.J., 1956, The regenerative responses of amputated limbs to delayed insertion into the body cavity, Anat. Rec., 126:283–297.

    Article  PubMed  CAS  Google Scholar 

  • Greene, L.A., and Tischler, A.S., 1976, Establishment of a noradrenergic clone line of rat adrenal pheochromocytoma cells which respond to nerve growth factor, Proc. Natl. Acad. Sci. USA, 73:2424–2428.

    Article  PubMed  CAS  Google Scholar 

  • Guyton, J.R., Rosenberg, R.D., Clowes, A.W., and Karnovsky, M.J., 1980, Inhibition of rat arterial smooth muscle cell proliferation by heparin. In vivo studies with anticoagulant and nonanticoagulant heparin, Circ. Res., 46:625–634.

    PubMed  CAS  Google Scholar 

  • Hamerman, D., Taylor, S., Kirschenbaum, I., Klagsbrun, M., Rainer, E.W., Ross, R., and Thomas, K.A., 1987, Growth factors with heparin binding affinity in human synovial fluid, Proc. Soc. Exp. Biol. Med.. 186:384–389.

    PubMed  CAS  Google Scholar 

  • Hauschka, P.V., Mavrakas, M.E., Iafrati, M.D., Doleman, S.E., and Klagsbrun, M., 1986, Growth factors in bone matrix: isolation of multiple types by affinity chromotography on heparin-sepharose, J. Biol. Chem., 261:12665–12674.

    PubMed  CAS  Google Scholar 

  • Hideaki, A., and Hiroyuki, I., 1988, A gradient of responsiveness to the growth-promoting activity of ZPA (zone of polarizing activity) in the chick limb bud. Develop. Biol.. 128:136–141.

    Article  Google Scholar 

  • Hoover, R.L., Rosenberg, R.D., Haering, W., and Karnovsky, M.J., 1980, Inhibition of rat arterial smooth muscle cell proliferation by heparin, Circ. Res., 47:578–583.

    PubMed  CAS  Google Scholar 

  • Inamura, T., and Mitsui, Y., 1987, Heparan sulfate and heparin as a potentiator or a suppressor of growth of normal and transformed vascular endothelial cells, Exp. Cell. Res., 172:92–100.

    Article  Google Scholar 

  • Kardami, E., Spector, D., and Srohman, R.C., 1988, Heparin inhibits skeletal muscle growth in vitro. Develop. Biol., 126:19–29.

    Article  PubMed  CAS  Google Scholar 

  • Lheureux, E., 1983, Replacement of irradiated epidermis by migration of non irradiated epidermis in the newt limb: the necessity of healthy epidermis for regeneration, J. Embryol. exp. Morph., 76:217–234.

    PubMed  CAS  Google Scholar 

  • Lobb, R.R., Harper, J.W., and Fett, J.W., 1986, Purification of heparin binding growth factors, Anal. Biochem., 154:1–14.

    Article  PubMed  CAS  Google Scholar 

  • Maciag, T., Mehlman, T., Friesel, R., and Schreiber, A.B., 1984, Heparin binds endothelial cell growth factor, the principal endothelial cell mitogen in bovine brain, Science. 225:932–934.

    Article  PubMed  CAS  Google Scholar 

  • Majack, R.A. , Cook, S.C., and Bornstein, P., 1986, Control of smooth muscle growth by composants of the extracellular matrix: autocrine role for thrombospondin, Proc. Natl. Acad. Sci. USA. 83:9050–9054.

    Article  PubMed  CAS  Google Scholar 

  • Malinin, T., and Deck, J.D., 1958, The effects of implantation of embryonic and tadpole tissues into adult frog limbs. I. Regeneration after amputation, J. Exp. Zool., 139:307–328.

    Article  PubMed  CAS  Google Scholar 

  • Mc Lachlan, J.C., Macintyne, J., Hume, D.D., and Smith, J., 1988, Direct demonstration of production of transforming growth factor activity by embryonic chick tissue, Experientia., 44:351–352.

    Article  CAS  Google Scholar 

  • Moscatelli, D., Presta, M., Joseph-Silverstein, J., and Rifkin, D.B., 1986, Both normal and tumor cells produce basic fibroblast growth factor, J. Cell Physiol.. 129:273–276.

    Article  PubMed  CAS  Google Scholar 

  • Neufeld, G., Gospodarowicz, D., Dodge, L., and Fujii, D.K., 1987, Heparin modulation of the neurotropic effects of acidic and basic fibroblast growth factors and nerve growth factors on PC12, J. Cell. Physiol.. 131:131–140.

    Article  PubMed  CAS  Google Scholar 

  • Orlidge, A., and D’Amore, P., 1986, Cell specific effects of glycosaminoglycans on the attachment and proliferation of vascular wall components, Microvasc. Res.. 31:41–53.

    Article  PubMed  CAS  Google Scholar 

  • Polezhaev, L.V., 1959, Restoration of regenerative ability of the extremities of axolotls after irradiation with roentgen rays, Dokl. Akad. Nauk. SSSR., 127:713–716.

    Google Scholar 

  • Polezhaev, L.V., and Ermakova, N.I., 1960, Restoration of regenerative capacity of the extremities of axolotls depressed by roentgen radiation, Dokl. Akad. Nauk. SSSR., 131:209–212.

    Google Scholar 

  • Polezhaev, L.V., and Faworina, W.N., 1935, Uber die Rolle des Epithels in den Anfanglichen Entwicklungsstadien einer Regenerationsanlage der Extremität beim Axolotl, Wilhelm Roux Arch., 133:701–727.

    Article  Google Scholar 

  • Polezhaev, L.V., Teplits, N.A., and Ermakova, N.I., 1961, Restoration of the regenerative property of extremities in axolotls inhibited by X-ray irradiation, by means of proteins, nucleic acids and lyophilized tissues, Dokl.Akad. Nauk. SSSR, 138:477–480.

    CAS  Google Scholar 

  • Polezhaev, L.V., Teplits, N.A., and Tuchkova, S.Ya., 1964, Restoration of regenerational capacity of extremities of axolotls after it has been suppressed by X-rays with the aid of nuclear acids, Dokl. Akad. Nauk. SSSR., 159:682–685.

    PubMed  CAS  Google Scholar 

  • Polezhaev, L.V., and Tuchkova, S.Ya, 1967, Restoration of regenerative power of axolotl limbs inhibited by X-ray irradiation with tissue heterografting, Dokl. Akad. Nauk. SSSR., 180:754–757.

    Google Scholar 

  • Reilly, C.F., Fritze, L., and Rosenberg, R.D., 1986, Heparin inhibition of smooth muscle cell proliferation: a cellular site of action, J. Cell. Physiol., 129:11–19.

    Article  PubMed  CAS  Google Scholar 

  • Revardel, J.L., and Chapron, C., 1975, Influence de la vascularization sur la regeneration des membres chez les larves d’ Urodèles. Nouvelle interprétation du rôle du système nerveux. C.R. Acad. Sc., 280:1409–1411.

    Google Scholar 

  • Richmond, M.J., and Pollack, E.D., 1983, Regulation of tadppole spinal nerve fiber growth by the regenerating limb blastema in tissue culture, J. Exp. Zool., 225:233–242.

    Article  PubMed  CAS  Google Scholar 

  • Rosenbaum, J., Tobelem, G., Molho, P., Barzy, T., and Caen, J.P., 1986, Modulation of endothelial cells growth induced by heparin, Cell. Biol. Int. Rep., 10:437–446.

    Article  PubMed  CAS  Google Scholar 

  • Rydel, R.E., and Greene, L.A., 1987, Acidic and basic fibroblast growth factors promote stable neurite outgrowth and neuronal differentiation in cultures of PC12 cells, J. Neurosc., 7:3639–3653.

    CAS  Google Scholar 

  • Schreiber, A.B., Kenney, J., Kowalski, J., Thomas, K.A., Gimenez-Gallego, G., Rios-Candelore, M., Di salvo, J., Barritault, D., Courty, J., Courtois, Y., Moenner, M., Loret, C., Burgess, W.H., Mehlman, T., Friesel, R., Johnson, W., and Maciag, T., 1985y, A unique family of endothelial cell polypeptide mitogens: endothelial cell growth factor, brain-derived acidic fibroblast growth factor and eye-derived growth factor II, J. Cell. Biol., 101:1623–1626.

    Article  PubMed  CAS  Google Scholar 

  • Schreiber, A.B., Kenney, J., Kowalski, J., Friesel, R., Mehlman, T., and Maciag, T., 1985, The interaction of endothelial cell growth factor with heparin: characterization by receptor and antibody recognition, Proc. Natl. Acad. Sci.USA., 82:6138–6142.

    Article  PubMed  CAS  Google Scholar 

  • Schubert, D., Ling, N., and Baird, A., 1987, Multiple influence of a heparin-binding growth factor on neuronal development, J. Cell Biol., 104:635–643.

    Article  PubMed  CAS  Google Scholar 

  • Schweigerer, L., Neufeld, G., Friedman, J., Abraham, J.A., and Fidder, J.C., 1987, Capillary endothelial cells express basic fibroblast growth factor, a mitogen that promotes their own growth, Nature, 325:257–259.

    Article  PubMed  CAS  Google Scholar 

  • Seed, J., and Hauschka, S.D., 1988, Clonal analysis of vertebrate myogenesis. Fibroblast growth factor (FGF)-dependent and FGF-independent muscle colony types during chick wing development, Develop. Biol., 128:40–49.

    Article  PubMed  CAS  Google Scholar 

  • Shing, Y., Folkman, J., Sullivan, R., Butterfield, C., Murray, J., and Klagsbrun, P., 1984, Heparin affinity: purification of a tumor-derived capillary endothelial cell growth factor, Science. 223:1296–1299.

    Article  PubMed  CAS  Google Scholar 

  • Singer, M., 1974, Neurotrophic control of limb regeneration in the newt, Ann. NY Acad. Sci., 228:308–322.

    Article  PubMed  CAS  Google Scholar 

  • Singer, M., 1978, On the nature of neurotrophic phenomenon in urodele limb regeneration, Am. Zool., 18:829–841.

    CAS  Google Scholar 

  • Smith, St D., and Crawford, G.L., 1969, Initiation of regeneration in adult Rana pipiens limbs by injection of homologous liver nuclear R.N.P., Oncogenesis, 23:299–307.

    CAS  Google Scholar 

  • Smith, A.R., and Wolpert, L., 1975, Nerves and angiogenesis in amphibian limb regeneration, Nature, 257:224–225.

    Article  PubMed  CAS  Google Scholar 

  • Stocum, D.L., and Dearlove, G., 1972, Epidermal-mesodermal interaction during morphogenesis of the limb regeneration blastema in larval salamander, J. Exp. Zool., 181:49–62.

    Article  Google Scholar 

  • Thornton, C.S., Mueller, S.N., and Levine, E.M., 1983, Human endothelial cells: use of heparin in cloning and longterm serial propagation, Science, 222:623–625.

    Article  PubMed  CAS  Google Scholar 

  • Todd, T.J., 1823, On the process of reproduction of the members of the aquatic salamander, Quart. J. Sci. Lit. Arts, 16:84–96.

    Google Scholar 

  • Togari, A., Baker, D., Dickens, G., and Guroff, G., 1983. The neurite promoting effect of fibroblast growth factor on PC12 cells, Biochem. Biophys. Res. Com., 114:1189–1193.

    Article  PubMed  CAS  Google Scholar 

  • Togari, A., Dickens, G., Kuzuya, H., and Guroff, G., 1985, The effect of fibroblast growth factor on PC12 cells, J. Neurose., 5:307–316.

    CAS  Google Scholar 

  • Uhlrich, S., Lagente, O., Choay, J., Courtois, Y., and Lenfant M. , 1986a, Structure activity relationships in heparin: stimulation of non vascular cells by a synthetic heparin pentasaccharide in cooperation with human acidic fibroblast growth factor, Biochem. Biophys. Res. Comm.. 139:728–732.

    Article  PubMed  CAS  Google Scholar 

  • Uhlrich, S., Lagente, O., Lenfant, M., and Courtois, Y., 1986b, Effect of heparin on the stimulation of non vascular cells by human acid and basic FGF, Biochem. Biophys. Res. Comm., 137:1205–1213.

    Article  PubMed  CAS  Google Scholar 

  • Wagner, J.A., and D’ Amore, P.A., 1986, Neurite outgrowth induced by an endothelial cell mitogen isolated from retina, J. Cell. Biol., 103:1363–1367.

    Article  PubMed  CAS  Google Scholar 

  • Wallace, H., 1981, “Vertebrate limb regeneration”, John Wiley & Sons, Chichester.

    Google Scholar 

  • Winkles, J.A., Friesel, R., Burgess, W.H., Howk, R., Mehlman, T., Weinstein, R., and Maciag, T., 1987, Human vascular smooth muscle cells both express and respond to hepar-in-binding growth factor I (endothelial cell growth factor), Proc. Natl. Acad. Sci. USA., 84:7124–7128.

    Article  PubMed  CAS  Google Scholar 

  • Wright, T.C., Johnstone, T.V., Castellot, J.J., and Karnovsky, M.J., 1985, Inhibition of rat cervical epithelial cell growth by heparin and its reversal by EGF, J. Cell. Physiol.. 125:499–506.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Laboratoire de Biologie des Facteurs de Croissance, Université de Lille, France

    B. Boilly

  2. Department of Biological Chemistry and Developmental Biology Center, University of California, Irvine, USA

    B. Boilly

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  1. B. Boilly
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Editors and Affiliations

  1. University of Athens, Athens, Greece

    V. Kiortsis  & S. Koussoulakos  & 

  2. University of Birmingham, Birmingham, UK

    H. Wallace

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© 1989 Plenum Press, New York

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Boilly, B. (1989). Production of Growth Factors by the Blastema During Limb Regeneration of Urodeles (Amphibia). In: Kiortsis, V., Koussoulakos, S., Wallace, H. (eds) Recent Trends in Regeneration Research. NATO ASI Series, vol 172. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-9057-2_8

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  • DOI: https://doi.org/10.1007/978-1-4684-9057-2_8

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