Skip to main content
Log in

Stability of positional identity of axolotl blastema cells in vitro

  • Original Articles
  • Published:
Roux's archives of developmental biology Aims and scope Submit manuscript

Summary

Previous grafting experiments have demonstrated that cells from non-contiguous positions within developing and regenerating limbs differ in a property referred to as positional identity. The goal of this study was to determine how long the positional identity of axolotl limb blastema cells is stable during culture in vitro. We have developed an assay for posterior positional properties such that blastema cells can be cultured and then grafted into anterior positions in host blastemas, to determine if they can stimulate supernumerary digit formation. We report that posterior blastema cells are able to maintain their positional identities for at least a week in culture. In addition, we observed that blastema cells are able to rapidly degrade collagenous substrates in vitro, a property that apparently distinguishes them from limb cells of other vertebrates. These results provide information regarding the time boundaries within which the positional properties of blastema cells can be studied and manipulated in vitro.

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

  • Bryant SV, Iten L (1974) The regulative ability of the limb regeneration blastema of Notophthalmus viridescens: Experiments in situ. Roux's Arch Dev Biol 174:90–101

    Google Scholar 

  • Bryant SV, Iten LE (1977) Intercalary and supernumerary regeneration in regenerating and mature limbs of Notophthalmus viridescens. J Exp Zool 202:1–16

    Google Scholar 

  • Bryant SV, French V, Bryant PJ (1981) Distal Regeneration and Symmetry. Science 212:993–1002

    Google Scholar 

  • Bryant SV, Gardiner DM, Muneoka K (1987) Limb development and regeneration. Am Zool 27:675–696

    Google Scholar 

  • Conn ME, Dearlove GE, Dresden MH (1979) Selection of a chemically defined medium for culturing adult newt forelimb regenerates. In Vitro 15:409–414

    Google Scholar 

  • Dollé P, Izpisua-Benmonte J-C, Falkenstein H, Renucci A, Duboule D (1989) Coordinate expression of the murine Hox-5 complex homoeobox-containing genes during limb pattern formation. Nature 342:767–772

    Google Scholar 

  • Dresden MH, Gross J (1970) The collagenolytic enzyme of the regenerating limb of the newt Triturus viridecens. Dev Biol 22:129–137

    Google Scholar 

  • Ferretti P, Brookes JP (1988) Culture of newt cells from different tissues and their expression of a regeneration-associated antigen. J Exp Zool 247:77–91

    Google Scholar 

  • Gardiner DM, Bryant SV (1989) Organization of positional information in the axolotl limb. J Exp Zool 251:47–55

    Google Scholar 

  • Gardiner DM, Gaudier C, Bryant SV (1992) Mouse limb bud cells respond to retinoic acid in vitro with reduced growth. J Exp Zool 263:406–413

    Google Scholar 

  • Gardiner DM, Muneoka K, Bryant SV (1986) The migration of dermal cells during blastema formation in axolotls. Dev Biol 118:488–493

    Google Scholar 

  • Grillo HC, Lapiere CM, Dresden MH, Gross J (1968) Collagenolytic activity in regenerating forelimbs of the adult newt (Triturus viridescens). Dev Biol 17:571–583

    Google Scholar 

  • Hayamizu TF, Bryant SV (1992) Retinoic acid respecifies limb bud cells in vitro. J Exp Zool: 263:423–429

    Google Scholar 

  • Hayamizu TF, Sessions SK, Wanck N, Bryant SV (1991) Effects of localized application of transforming growth factor β1 on developing chick limbs. Dev Biol 145:164–173

    Google Scholar 

  • Honig LS (1983) Polarizing activity of the avian limb examined on a cellular basis. In: Fallon JF, Caplan AI (eds) Limb development and regeneration. A R Liss, Inc, New York, pp 99–108

    Google Scholar 

  • Izpisúa-Belmonte J-C, Tickle C, Dollé P, Wolpert L, Duboule D (1991) Expression of the homebox Hox-4 genes and the specification of position in chick wing development. Nature 350: 585–589

    Google Scholar 

  • Jabaily JA, Blue P, Singer M (1982) The culturing of dissociated newt forelimb regenerate cells. J Exp Zool 219:67–73

    Google Scholar 

  • MacCabe AB, Gasseling MT, Saunders JW (1973) Spatiotemporal distribution of mechanisms that control outgrowth and anteroposterior polarization of the limb bud in the chick embryo. Mechanism Ageing Dev 2:1–12

    Google Scholar 

  • Muneoka K, Bryant SV (1984) Cellular contribution to supernumerary limbs in the axolotl, Ambystoma mexicanum. Dev Biol 105:166–178

    Google Scholar 

  • Muneoka K, Holler-Dinsmore G, Bryant SV (1986) Intrinsic control of regenerative loss in Xenopus laevis limbs. J Exp Zool 240:47–54

    Google Scholar 

  • Muneoka K, Holler-Dinsmore GV, Bryant SV (1985) A quantitative analysis of regeneration from chimaeric limb stumps in the axolotl. J Embryol Exp Morphol 90:1–12

    Google Scholar 

  • Nohno T, Noji S, Koyama E, Ohyama K, Myokai F, Kuroiwa A, Saito T, Taniguchi S (1991) Involvement of the Chox-4 chicken homeobox genes in determination of anteroposterior axial polarity during limb development. Cell 64:1197–1205

    Google Scholar 

  • Shi C, Muneoka K (1992) Position-specific growth of mouse limb bud cells in vitro. Dev Biol 151:9–17

    Google Scholar 

  • Tank PW, Carlson BM, Connelly TG (1976) A staging system for forelimb regeneration in the axolotl, Ambystoma mexicanum. J Morphol 150:117–128

    Google Scholar 

  • Tank PW, Connelly TG, Bookstein FL (1985) Cellular behavior in the anteroposterior axis of the regenerating forelimb of the axolotl, Ambystoma mexicanum. Dev Biol 109:215–223

    Google Scholar 

  • Tickle C (1981) The number of polarizing region cells required to specify additional digits in the developing chick wing. Nature 289:295–298

    Google Scholar 

  • Tickle C, Alberts B, Wolpert L, Lee J (1982) Local application of retinoic acid to the limb bond [sic] mimics the action of the polarizing region. Nature 296:564–566

    Google Scholar 

  • Tickle C, Summerbell D, Wolpert L (1975) Positional signalling and specification of digits in chick limb morphogenesis. Nature 254:199–202

    Google Scholar 

  • Yokouchi Y, Sasaki H, Kuroiwa A (1991) Homeobox gene expression correlated with the bifurcation process of limb cartilage development. Nature 353:443–445

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Correspondence to: S.V. Bryant

Rights and permissions

Reprints and permissions

About this article

Cite this article

Groell, A.L., Gardiner, D.M. & Bryant, S.V. Stability of positional identity of axolotl blastema cells in vitro. Roux's Arch Dev Biol 202, 170–175 (1993). https://doi.org/10.1007/BF00365307

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00365307

Key words

Navigation