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In vivo inhibition of programmed cell death by local administration of FGF-2 and FGF-4 in the interdigital areas of the embryonic chick leg bud

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Abstract

The formation of the digits in amniote vertebrates is accompanied by a massive degeneration process that accounts for the disappearance of the interdigital mesenchyme. The establishment of these areas of interdigital cell death (INZs) is concomitant with the flattening of the apical ectodermal ridge (AER), but a possible causal relationship between these processes has not been demonstrated. Recent studies have shown that the function of the AER can be substituted for by implantation of beads bearing either FGF-2 or FGF-4 into the apical mesoderm of the early limb bud. According to these observations, if the onset of INZs is triggered by the cessation of the AER function, local administration of FGFs to the interdigital tissue prior to cell death should delay or inhibit interdigit degeneration. In the present study we have confirmed this prediction. Implanting Affi-gel blue or heparin beads pre-absorbed with either FGF-2 or FGF-4 into the interdigital tissue of the chick leg bud in the stages prior to cell death stimulates cell proliferation and causes the formation of webbed digits. Vital staining with neutral red confirmed an intense temporal inhibition of interdigital cell death after FGF treatment. This inhibition of interdigital cell death was not accompanied by modifications in the pattern of expression of Msx-1 or Msx-2 genes, which in normal development display a domain of expression in the interdigital tissue preceding the onset of degeneration.

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References

  • Coelho CND, Sumoy L, Rodgers BJ, Davidson DR, Hill R, Upholt WB, Kosher RA (1991a) Expression of the chicken homeobox-containing gene GHox-8 during embryonic chick limb development. Mech Dev 34:143–154

    Google Scholar 

  • Coelho CND, Krabbenhoft KM, Upholt WB, Fallon JF, Kosher RA (1991b) Altered expression of the chicken homeobox-containing genes GHox-7 and GHox-8 in the limb buds of limbless mutant chick embryos. Development 113:1487–1493

    Google Scholar 

  • Cohn MJ, Izpisúa-Belmonte JC, Abud H, Heath JK, Tickle C (1995) Fibroblast growth factors induce additional limb development from the flank of chick embryos. Cell 80:739–746

    Google Scholar 

  • Crossley PH, Martin GR (1995) The mouse Fgf8 gene encodes a family of polypeptides and is expressed in regions that direct outgrowth and patterning in the developing embryo. Development 121:439–451

    Google Scholar 

  • Dealy CN, Kosher RA (1995) Studies on insulin-like growth factor-I and insulin in chick limb morphogenesis. Dev Dyn 202:67–79

    Google Scholar 

  • Fallon JF, Kelley RO (1977) Ultrastructural analysis of the apical ectodermal ridge during vertebrate limb morphogenesis. II. Gap junctions as distinctive ridge structures common to birds and mammals. J Embryol Exp Morphol 41:223–232

    Google Scholar 

  • Fallon JF, Lopez A, Ros MA, Savage MP, Olwin BB, Simandl BK (1994) FGF-2: apical ridge growth signal for chick limb development. Science 264:104–107

    Google Scholar 

  • Francis PH, Richardson MK, Brickell PM, Tickle C (1994) Bone morphogenetic proteins and a signalling pathway that controls patterning in the developing chick limb. Development 120: 209–218

    Google Scholar 

  • Gañan Y, Macias D, Hurle JM (1994) Pattern regulation in the chick autopodium at advanced stages of embryonic development. Dev Dyn 199:64–72

    Google Scholar 

  • Hamburger V, Hamilton H (1951) A series of normal stages in the development of the chick embryo. J Morphol 88:49–92

    Google Scholar 

  • Hill RE, Jones PF, Rees AR, Sime CM, Justice MJ, Copeland NG, Jenkins NA, Graham E, Davidson DR (1989) A new family of mouse homeobox-containing genes: Molecular structure, chromosomal location, and developmental expression of Hox 7.1. Genes Dev 3:26–37

    Google Scholar 

  • Hinchliffe JR, Ede DA (1973) Cell death and development of limb form and skeletal pattern in normal and wingless (ws) chick embryos. J Embryol Exp Morphol 30:753–772

    Google Scholar 

  • Hurle JM (1988) Cell death in developing systems. Methods Achiev Exp Pathol 13:55–86

    Google Scholar 

  • Hurle JM, Gañan Y (1986) Interdigital tissue chondrogenesis induced by surgical removal of the ectoderm in the embryonic chick leg bud. J Embryol Exp Morphol 94:231–244

    Google Scholar 

  • Hurle JM, Gañan Y (1987) Formation of extra digits induced by surgical removal of the apical ectodermal ridge of the chick embryo leg bud in the stages previous to the onset of interdigital cell death. Anat Embryol 176:393–399

    Google Scholar 

  • Hurle JM, Gañan Y, Macias D (1989) Experimental analysis of the in vivo chondrogenic potential of the interdigital mesenchyme of the chick leg bud subjected to local ectodermal removal. Dev Biol 132:368–374

    Google Scholar 

  • Hurle JM, Macias D, Gañan Y, Ros MA, Fernandez-Teran MA (1991) The interdigital spaces of the chick leg bud as a model for analysing limb morphogenesis and cell differentiation. In: Hinchliffe JR, Hurle JM, Summerbell D (eds) Developmental patterning of the vertebrate limb. Plenum Press, New York, pp 249–260

    Google Scholar 

  • Lee KKH, Li FCH, Yung WT, Kung JLS, Ng JL, Cheah KSE (1994) Influence of digits, ectoderm, and retinoic acid on chondrogenesis by mouse interdigital mesoderm in culture. Dev Dyn 201:297–309

    Google Scholar 

  • Lyons KM, Pelton RW, Hogan LM (1990) Organogenesis and pattern formation in the mouse: RNA distribution patterns suggest a role for bone morphogenetic protein-2A (BMP-2A). Development 109:833–844

    Google Scholar 

  • MacCabe JA, Blaylock RL, Latimer JL, Pharris LJ (1991) Fibroblast growth factor and culture in monolayer rescue mesoderm cells destined to die in th developing avian wing. J Exp Zool 257:208–213

    Google Scholar 

  • Macias D, Gañan Y, Hurle JM (1992) Interdigital chondrogenesis and extra digit formation in the duck leg bud subjected to local ectoderm removal. Anat Embryol 186:27–32

    Google Scholar 

  • Mima T, Ohuchi H, Noji S, Mikawa T (1995) FGF can induce outgrowth of somatic mesoderm both inside and outside of limb-forming regions. Dev Biol 167:617–620

    Google Scholar 

  • Mori C, Nakamura N, Kimura S, Irie H, Takigawa T, Shiota K (1995) Programmed cell death in the interdigital tissue of the fetal mouse limb is apoptosis with DNA fragmentation. Anat Rec 242:103–110

    Google Scholar 

  • Myokai F, Washio N, Asahara Y, Yamaai T, Tanda N, Ishikawa T, Aoki S, Kurihara H, Murayama Y, Saito T, Matsumoto K, Nakamura T, Noji S, Nohno T (1995) Expression of hepatocyte growth factor gene during chick limb development. Dev Dyn 202:80–90

    Google Scholar 

  • Niswander L, Martin GR (1992) Fgf-4 expression during gastrulation, myogenesis, limb and tooth development in the mouse. Development 114:755–768

    Google Scholar 

  • Niswander L, Tickle C, Vogel A, Booth I, Martin G (1993) FGF-4 replaces the apical ectodermal ridge and directs outgrowth and patterning of the limb. Cell 75:1–20

    Google Scholar 

  • Niswander L, Jeffrey S, Martin G, Tickle C (1994) A positive feedback loop coordinates growth and patterning in the vertebrate limb. Nature 371:609–612

    Google Scholar 

  • Oppenheim RW, Schwartz LM, Shatz CJ (1992) Neuronal death, a tradition of dying. J Neurobiol 23:1111–1115

    Google Scholar 

  • Oppenheim RW, Houenou LJ, Johnson JE, Lin L-FH, Li L, Lo AC, Newsome AL, Prevette DM, Wang S (1995) Developing motor neurons rescued from programmed and axotomy-induced cell death by GDNF. Nature 373:344–346

    Google Scholar 

  • Orr-Urtreger A, Givol D, Yayon A, Yarden Y, Lonai P (1991) Developmental expression of two murine fibroblast growth factor receptors, flg and bek. Development 113:1419–1434

    Google Scholar 

  • Pautou M-P (1978) Cessation de l'activité de la crête apicale ectodermique au cours de la morphogenèse de l'autopode chez l'embryon de poulet. Analyse histologique. Arch Biol (Bruxelles) 89:27–66

    Google Scholar 

  • Raynaud A, Kan P, Bouche G, Duprat A-M (1995) Fibroblast growth factor (FGF-2) and delayed involution of the posterior limbs of the slow-worm embryo (Anguis fragilis, L). CR Acad Sci III 318:573–578

    Google Scholar 

  • Robert B, Lyons G, Simandl BK, Kuroiwa A, Buckingham M (1991) The apical ectodermal ridge regulates Hox-7 and Hox-8 gene expression in developing chick limb buds. Genes Dev 5:2363–2374

    Google Scholar 

  • Ros MA, Lyons G, Kosher RA, Upholt WB, Coelho CND, Fallon JF (1992) Apical ridge dependent and independent mesodermal domains of GHox-7 and GHox-8 expression in chick limb buds. Development 116:811–818

    Google Scholar 

  • Ros MA, Macias D, Fallon JF, Hurle JM (1994) Formation of extra digits in the interdigital spaces of the chick leg bud is not preceded by changes in the expression of Msx and Hoxd genes. Anat Embryol 190:375–382

    Google Scholar 

  • Rowe RA, Fallon JF (1982) The proximodistal determination of skeletal parts in the developing chick leg. J Embryol Exp Morphol 68:1–7

    Google Scholar 

  • Rowe DA, Cairns JM, Fallon JF (1982) Spatial and temporal patterns of cell death in limb bud mesoderm after apical ectodermal ridge removal. Dev Biol 93:83–91

    Google Scholar 

  • Ruberte E, Friederich V, Morris-Kay G, Chambon P (1992) Differential distribution patterns of CRABP I and CRAPB II transcripts during mouse embryogenesis. Development 115: 973–987

    Google Scholar 

  • Ruit KG, Elliot JL, Osborne PA, Yan Q, Snider WD (1992) Selective dependence of mammalian dorsal root ganglion neurons on nerve growth factor during embryonic development. Neuron 8:573–587

    Google Scholar 

  • Saunders JW Jr (1948) The proximo-distal sequence of origin of limb parts of the chick wing and the role of ectoderm. J Exp Zool 108:363–404

    Google Scholar 

  • Savage MP, Fallon JF (1995) FGF-2 mRNA and its antisense message are expressed in a developmentally specific manner in the chick limb bud and mesonephros. Dev Dyn 202:343–353

    Google Scholar 

  • Savage MP, Hart CE, Riley BB, Sasse J, Olwin BB, Fallon JF (1993) Distribution of FGF-2 suggests it has a role in chick limb bud growth. Dev Dyn 198:159–170

    Google Scholar 

  • Solursh M (1984) Ecotderm as a determinant of early tissue pattern in the limb bud. Cell Differ 15:17–24

    Google Scholar 

  • Streck RD, Wood TL, Hsu M-S, Pintar JE (1992) Insulin-like growth factor I and II and insulin-like growth factor binding protein-2 RNAs are expressed in adjacent tissues within rat embryonic and fetal limbs. Dev Biol 151:586–596

    Google Scholar 

  • Summerbell D (1974) A quantitative analysis of the effect of excision of the AER from the chick limb bud. J Embryol Exp Morphol 32:651–660

    Google Scholar 

  • Suzuki HR, Padanilam BJ, Vitale E, Ramirez F, Solursh M (1991) Repeating developmental expression of GHox-7, a novel homeobox-containing gene in the chick limb morphogenesis. Dev Biol 148:375–388

    Google Scholar 

  • Suzuki HR, Sakamoto H, Yoshida T, Sugimura T, Terada M, Solursh M (1992) Localization of Hst1 transcripts to the apical ectodermal ridge in the mouse embryo. Dev Biol 150:219–222

    Google Scholar 

  • Tickle C, Eichele G (1994) Vertebrate limb development. Annu Rev Cell Biol 10:121–152

    Google Scholar 

  • Toné S, Tanaka S, Kato Y (1988) The cell cycle and cell population kinetics in the programmed cell death in the limb-buds of normal and 5-bromodeoxyuridine-treated chick embryos. Dev Growth Differ 30:261–270

    Google Scholar 

  • Wilkie AOM, Slaney SF, Oldridge M, Poole MD, Ashworth GJ, Hockley AD, Hayward RD, David DJ, Pulley LJ, Rutland P, Malcolm S, Winter RM, Reardon W (1995a) Apert syndrome results from localized mutations of FGFR2 and is allelic with Crouzon syndrome. Nature Genetics 9:165–172

    Google Scholar 

  • Wilkie AOM, Morris-Kay GM, Jones GM, Heath JK (1995b) Functions of fibroblast growth factors and their receptors. Curr Biol 5:500–507

    Google Scholar 

  • Yang Y, Niswander L (1995) Interaction between the signalling molecules WNT7a and SHH during vertebrate limb development: dorsal signals regulate anteroposterior patterning. Cell 80:939–947

    Google Scholar 

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Authors and Affiliations

  1. Departamento de Ciencias Morfológicas, Universidad de Extremadura, E-06071, Badajoz, Spain

    Domingo Macias & Yolanda Gañan

  2. Departamento de Anatomía y Biología Celular, Facultad de Medicina, Universidad de Cantabria, Polígono de Cazoña, E-39011, Santander, Spain

    Maria A. Ros & Juan M. Hurle

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  1. Domingo Macias
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  2. Yolanda Gañan
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  3. Maria A. Ros
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  4. Juan M. Hurle
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Macias, D., Gañan, Y., Ros, M.A. et al. In vivo inhibition of programmed cell death by local administration of FGF-2 and FGF-4 in the interdigital areas of the embryonic chick leg bud. Anat Embryol 193, 533–541 (1996). https://doi.org/10.1007/BF00187925

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