Skip to main content
Log in

Extracellular matrix modifications in the interdigital spaces of the chick embryo leg bud during the formation of ectopic digits

  • Original Article
  • Published:
Anatomy and Embryology Aims and scope Submit manuscript

Abstract

In previous studies we have observed that the interdigital mesenchyme of the chick leg bud, in the stages preceding the onset of cell death, retains a significant regulatory potential, forming ectopic extra digits under a variety of surgical manipulations. Most evidence suggests that interdigital extra digits are caused by the abolition of local antichondrogenic effects operating in the interdigital spaces under normal conditions rather than by modifications of the signalling mechanisms accounting for the normal patterning of the digits in early stages of development. The interdigital spaces exhibit a complex scaffold of extracellular matrix with well-defined domains of spatial distribution of type I and type VI collagens, tenascin, fibronectin, laminin and elastic matrix components that have been proposed to play a role in the establishment of the non-chondrogenic fate of the interdigital tissue in situ. In an attempt to analyze this possible role of the interdigital extracellular matrix (ECM), in the present work we have studied changes in the pattern of ECM distribution associated with the formation of extra digits. Extra digits were induced by making a T-cut in the third interdigital space of the leg but of stage 29 HH chick embryos. Subsequent modifications of the ECM were detected immunohistochemically in whole-mount specimens using laser confocal microscopy. Our results reveal that in the first hours after the operation, changes in the ECM apparently related to the healing of the wound cause a significant reorganization of the normal ECM scaffold of the interdigit. In addition, chondrogenesis of the interdigital tissue is preceded by disappearance of elastin fibers in the interdigital mesenchyme subjacent to the wound and by an intense deposition of tenascin. Tenascin deposition and loss of the elastin fibrillar scaffold were also observed preceding chondrogenesis in fragments of interdigital tissue explanted to culture conditions. The significance of these observations in relation to the establishment of the skeletal elements of the autopodium is discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Bressan GM, Daga-Gordini D, Colombatti A, Castellani I, Marigo V, Volpin D (1993) Emilin, a component of elastic fibers preferentially located at the elastin-microfibrils interface. J Cell Biol 121:201–212

    Google Scholar 

  • Chuong C-M, Widelitz RB, Jiang T-X, Abbott UK, Lee Y-S, Chen H-M (1993) Roles of adhesion molecules NCAM and tenascin in limb skeletogenesis: analysis with antibody perturbation, exogenous gene expression, talpid 2mutants and activin stimulation. In: Fallon JF, Goetinck PF, Kelley RO, Stocum DL (eds) Limb development and regeneration, part B. Wiley-Liss, New York, pp 465–474

    Google Scholar 

  • Davis EC (1993) Stability of elastin in the developing mouse aorta: a quantitative radioautographic study. Histochemistry 100:17–26

    Google Scholar 

  • Downie SA, Newman SA (1994) Morphogenetic differences between fore and hind limb precartilage mesenchyme: relation to mechanisms of skeletal pattern formation. Dev Biol 162:195–208

    Google Scholar 

  • Ffrench-Constant C, Van De Water L, Dvorak HF, Hynes RO (1989) Reappearance of an embryonic pattern of fibronectin splicing during wound healing in the adult rat. J Cell Biol 109:903–914

    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 

  • Garcia-Martinez V, Macias D, Gañan Y, Garcia-Lobo JM, Francia MV, Fernandez-Teran MA, Hurle JM (1993) Internucleosomal DNA fragmentation and programmed cell death (apoptosis) in the interdigital tissue of the embryonic chick leg bud. J Cell Sci 106:201–208

    Google Scholar 

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

    Google Scholar 

  • Hinchliffe JR, Horder TJ (1993) Lessons from extradigits. In: Fallon JF, Goetinck PF, Kelley RO, Stocum DL (eds) Limb development and regeneration, part B. Wiley-Liss, New York, pp 113–126

    Google Scholar 

  • Holzenberger M, Lièvre CA, Robert L (1993) Tropoelastin gene expression in the developing vascular system of the chicken: an in situ hybridization study. Anat Embryol 188:481–492

    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 extradigits 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 (1989a) 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, Hinchliffe JR, Ros MA, Critchlow MA, Genis-Galvez JM (1989b) The extracellular matrix architecture relating to myotendinous pattern formation in the distal part of the developing chick limb: an ultrastructural, histochemical and immunocytochemical analysis. Cell Differ 27:103–120

    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 analyzing limb morphogenesis and cell differentiation. In: Hinchliffe JR, Hurle JM, Summerbell D (eds) Developmental patterning of the vertebrate limb. Plenum Press, New York, pp 249–259

    Google Scholar 

  • Hurle JM, Corson G, Daniels K, Reiter R, Sakai LY, Solursh M (1994) Elastin exhibits a distinctive temporal and spatial pattern of distribution in the developing chick limb in association with the establishment of the cartilaginous skeleton. J Cell Sci 107:2623–2634

    Google Scholar 

  • Johnson DJ, LaBourne J, Rabinovitch M, Keeley FW (1993) Relative efficiency of incorporation of newly snythesized elastin and collagen into aorta, pulmonary artery and pulmonary vein of growing pigs. Connect Tissue Res 29:213–221

    Google Scholar 

  • Lee KKH, Chan WY, Sze LY (1993) Histogenetic potential of rat hind-limb interdigital tissues prior to and during the onset of programmed cell death. Anat Rec 236:568–572

    Google Scholar 

  • Lee KHK, 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 

  • Linsenmayer TF, Hendrix MJC, Little CD (1979) Production and characterization of a monoclonal antibody to chicken type I collagen. Proc Nat Acad Sci USA 76:3703–3707

    Google Scholar 

  • Macias D, Gañan Y (1991) The role of the polarizing zone in the pattern of experimental chondrogenesis in the chick embryo interdigital space. Int J Dev Biol 35:63–67

    Google Scholar 

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

    Google Scholar 

  • Mackie EJ, Thesleff I, Chiquet-Ehrismann R (1987) Tenascin is associated with chondrogenesis and osteogenic differentiation in vivo and promotes chondrogenesis in vitro. J Cell Biol 105:2569–2579

    Google Scholar 

  • Mackie EJ, Halfter W, Liverani D (1988) Induction of tenascin in healing wounds. J Cell Biol 107:2757–2767

    Google Scholar 

  • McCullagh JJ, Wilson DJ (1993) Antero-posterior skeletal patterning is not dependent on continuity of the apical ectodermal ridge in the chick wing bud. Anat Embryol 188:371–379

    Google Scholar 

  • Quarto R, Dozin B, Bonaldo P, Candedda R, Colombatti A (1993) Type VI collagen expression is upregulated in the early events of chondrocyte differentiation. Development 117:245–251

    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 the msx and hoxd genes. Anat Embryol 190:375–382

    Google Scholar 

  • Rosenblom J, Abrams WR, Mecham R (1993) Extracellular matrix 4: the elastic fiber. FASEB J 7:1208–1218

    Google Scholar 

  • Sage H, Gray WR (1979) Studies on the evolution of elastin. I. Phylogenetic distribution. Comp Biochem Physiol [B] 64:313–327

    Google Scholar 

  • Sakai LY, Keene DR, Engvall E (1986) Fibrillin, a new 350-kD glycoprotein, is a component of extracellular microfibrils. J Cell Biol 103:2499–2509

    Google Scholar 

  • Solursh M, Singley CT, Reiter RS (1981) The influence of epithelia on cartilage and loose connective tissue formation by limb mesenchyme cultures. Dev Biol 86:471–482

    Google Scholar 

  • Whitby DJ, Longaker MT, Harrison MR, Adzick NS, Ferguson MWJ (1991) Rapid epithelialisation of fetal wounds is associated with the early deposition of tenascin. J Cell Sci 99:pp 583–586

    Google Scholar 

  • Wright E, Hargrave MR, Christiansen J, Cooper L, Kun J, Evan T, Gangadharan U, Greenfield A, Koopman P (1995) The Sry-related gene Sox9 is expressed during chondrogenesis in mouse embryos. Nature Genetics 9:15–20

    Google Scholar 

  • Zanetti N, Solursh M (1986) Epithelial effects on limb chondrogenesis involve extracellular matrix and cell shape. Dev Biol 113:110–118

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hurle, J.M., Colombatti, A. Extracellular matrix modifications in the interdigital spaces of the chick embryo leg bud during the formation of ectopic digits. Anat Embryol 193, 355–364 (1996). https://doi.org/10.1007/BF00186692

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00186692

Key words

Navigation