Biochemical Pathways Involved in the Respecification of Pattern by Retinoic Acid

  • Malcolm Maden
  • Denis Summerbell
Part of the NATO ASI Series book series (NSSA, volume 172)


We describe here experiments designed to further our understanding of the ways in which cells of the developing chick limb bud and the regenerating axolotl limb detect and respond to retinoids during the process of pattern respecification. The binding proteins for retinoic acid (CRABP) and retinol (CRBP) have been identified and quantified in the chick and axolotl. In an attempt to determine the role of CRABP the potency (ability to respecify pattern) of a range of retinoid analogues was correlated with their binding affinities to CRABP. Analogues that were inactive did not bind to CRABP and with the exception of one analogue, arotinoid, the converse was also true. Immunolocalisation of CRABP in the chick limb bud showed that it was concentrated in the progress zone at the tip of the limb and distributed in a graded form across the anteroposterior axis with the high point on the anterior side. Immunolocalisation of CRBP showed that it was restricted solely to the posterior side of the limb bud, the converse of CRABP. The relationship between these distributions and the endogenous gradient of retinoic acid is discussed. Finally, experiments on retinoic acid-induced changes in proteins, particularly protein phosphorylation are described and it is suggested that these may be casual in the respecification of pattern.


Retinoic Acid Retinyl Palmitate Anteroposterior Axis Noic Acid Chick Limb 


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  1. Cohen, P., 1985, The role of protein phosphorylation in the hormonal control of enzyme activity. Eur. J. Biochem., 151:439–448.PubMedCrossRefGoogle Scholar
  2. Chytil, F. and Ong, D.A., 1984, Cellular retinol-binding proteins in: “The Retinoids”, M.B. Sporn, A.B. Roberts, S. Goodman, eds., Academic Press, Orlando.Google Scholar
  3. Eichele, G., Tickle, C. and Alberts, B.W., 1985, Studies on the mechanism of retinoid-induced pattern duplications in the early chick limb bud: temporal and spatial aspects. J. Cell Biol., 101:1913–1920.PubMedCrossRefGoogle Scholar
  4. Fell, H.B. and Mellanby, E., 1952, The effect of hypervitaminosis A on embryonic limb-bones cultivated in vitro. J. Physiol., 116:320–349.PubMedGoogle Scholar
  5. Fell, H.B. and Mellanby, E., 1953, Metaplasia produced in cultures of chick ectoderm by high Vitamin A. J. Physiol., 119:470–488.PubMedGoogle Scholar
  6. Giguere, V., Ong, E.S., Segui, P. and Evans, R.M., 1987, Identification of a receptor for the morphogen retinoic acid. Nature, 330:624–629.PubMedCrossRefGoogle Scholar
  7. Horton, W.E., Yamada, Y. and Hassell, J.R., 1987, Retinole acid rapidly reduces cartilage matrix synthesis by altering gene transcription in chondrocytes. Devel. Biol., 123:508–516.CrossRefGoogle Scholar
  8. Keeble, S. and Maden, M., 1986, Retinoic acid-binding protein in the axolotl: distribution in mature tissues and time of appearance during limb regeneration. Devel. Biol., 117:435–441.CrossRefGoogle Scholar
  9. Kim, W.S. and Stocum, D.L., 1986, Retinoic acid modifies positional memory in the anteroposterior axis of regenerating limbs. Devel. Biol., 114:170–179.CrossRefGoogle Scholar
  10. Lotan, R., 1980, Effects of Vitamin A and its analogues (retinoids) on normal and neoplastic cells. Biochim. Biophvs. Acta. 605:33–91.Google Scholar
  11. Maden, M., 1982, Vitamin A and pattern formation in the regenerating limb. Nature, 295:672–675.PubMedCrossRefGoogle Scholar
  12. Maden, M., 1983a, The effect of vitamin A on limb regeneration in Rana temporaria. Devel. Biol., 98:409–416.Google Scholar
  13. Maden, M., 1983b, The effect of vitamin A on the regenerating axolotl limb. J. Embrvol. exp. Morph., 77:273–295.Google Scholar
  14. Maden, M., 1984, Does vitamin A act on pattern formation via the epidermis or the mesoderm? J. exp. Zool., 230:387–392.CrossRefGoogle Scholar
  15. Maden, M. and Keeble, S., 1987, The role of cartilage and fibronectin during respecification of pattern induced in the regenerating amphibian limb by retinoic acid. Differentiation, 36:175–184.PubMedCrossRefGoogle Scholar
  16. Maden, M. and Summerbell, D., 1986, Retinoic acid-binding protein in the chick limb bud: identification at various developmental stages and binding affinities of various retinoids. J. Embrvol. exp. Morph., 97:239–250.Google Scholar
  17. Maden, M., Keeble, S. and Cox, R.A., 1985, The characteristics of local application of retinoic acid to the regenerating axolotl limb. Roux’s Arch. Dev. Biol., 194:228–235.Google Scholar
  18. Niazi, I.A. and Saxena, S., 1978, Abnormal hind limb regeneration in tadpoles of the toad, Bufo andersoni, exposed to excess vitamin A. Folia Biol. (Krakow), 26:3–8.Google Scholar
  19. Petkovich, M., Brand, N.J., Krust, A. and Chambon, P., 1987, A human retinoic acid receptor which belongs to the family of nuclear receptors. Nature, 330:444–450.PubMedCrossRefGoogle Scholar
  20. Porter, S.B., Ong, D.E., Chytil, F. and Orgebin-Crist, M-C., 1985, Localization of cellular retinol-binding protein and cellular retinoic acid-binding protein in the rat testis and epididymis. J. Androl., 6:197–212.PubMedGoogle Scholar
  21. Roberts, A.B., and Sporn, M., 1984, Cellular biology and biochemistry of the retinoids, in: “The Retinoids”, M.B. Sporn, A.B. Roberts, S. Goodman, eds., Academic Press, Orlando.Google Scholar
  22. Scadding, S.R., Vitamin A inhibits tail regeneration. Canad. J. Zool., 65:457–459.CrossRefGoogle Scholar
  23. Sharma, K.K. and Anton, H.J., 1986, Biochemical and ultrastructural studies of vitamin A induced proximalisation of limb regeneration in axolotl, in: “Progress in Developmental Biology”, H.C. Slavkin ed., Alan R. Liss, New York.Google Scholar
  24. Slack, J.M.W., 1983, Regional differences of protein synthesis in the limb regeneration blastema of the axolotl. Prog. in Clin. Biol. Res., 110A:557–563.Google Scholar
  25. Summerbell, D., 1983, The effect of local application of retinoic acid to the anterior margin of the developing chick limb bud. J. Embrvol. exp. Morph., 78:269–289.Google Scholar
  26. Summerbell, D. and Harvey, F., 1983, Vitamin A and the control of pattern in developing limbs. Prog. Clin. Biol, Res., 110A:109–118.Google Scholar
  27. Takase, S., Ong, D.E. and Chytil, F., 1986, Transfer of reti-noic acid from its complex with cellular retinoic acid-binding protein to the nucleus. Arch. Biochem. Biophvs.. 247:328–334.CrossRefGoogle Scholar
  28. Thaller, C. and Eichele, G., 1987, Identification and spatial distribution of retinoids in the developing chick limb bud. Natureu, 327:625–628.CrossRefGoogle Scholar
  29. Tickle, C., Alberts, B., Wolpert, L. and Lee, J., 1982, Local application of retinoic acid to the limb bud mimics the action of the polarizing region. Nature, 296:564–566.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1989

Authors and Affiliations

  • Malcolm Maden
    • 1
  • Denis Summerbell
    • 1
  1. 1.Limb Development GroupNational Institute for Medical ResearchMill Hill, LondonUK

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