Advertisement

Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Cyclic nucleotide metabolism during amphibian forelimb regeneration

II. The protein kinases

  • 28 Accesses

  • 4 Citations

Summary

The hypothesis that cAMP mediates neural and endocrine influences on limb regeneration was examined by studying the protein kinases in regenerating limb tissues. Since these enzymes are the vehicles through which cAMP acts intracellularly, an understanding of changes in their concentrations and behaviors during regeneration can be instrumental in elucidating the role of cAMP in this process. Mean activities oscillated throughout regeneration with maximal activities being observed during the mid-late bud stage. The phosphorylation of histone, added to the assay, varied with the stage of regeneration-greatest activity occurring during the early bud stage and very weak activity during the palette and early digital stages. Histone actually appeared to inhibit endogenous phosphorylation during dedifferentiation. In addition, cAMP demonstrated different degrees of enhancement of histone phosphorylation during regeneration-producing its greatest effect at the palette stage and having the least effect at the early bud stage. The results of this study suggest that changes in the absolute amounts of protein kinase are probably not significant in the regulation of regeneration. In addition, the variable acceptability of histone as an exogenous substrate and the variations in the cAMP effects on phosphorylation suggest that physiological changes are occurring in which cAMP might play a significant role. In particular, these data suggest that cAMP might be instrumental in influencing events associated with differentiation and morphogenesis.

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

References

  1. Babich GL, Foret JE (1973) Effects of dibutyryl cyclic AMP and related compounds on newt limb regeneration blastemas in vitro. II.14C-leucine incorporation. Oncology 28:88–95

  2. Berridge, MJ (1975) Control of cell division: a unifying hypothesis. J Cyclic Nucl Res 1:305–320

  3. Carroll JM, Sicard RE (1980) Cyclic nucleotide metabolism during amphibian forelimb regeneration. I. The cyclic AMP phosphodiesterases. Wilhelm Roux's Arch 189:107–110

  4. Danehy DF, Sicard RE (1978) Regional differences in soluble proteins of regenerating limbs of the adult newt,Notophthalmus viridescens. J Cell Biol 79:337a

  5. Dearlove GE, Stocum DL (1974) Denervation-induced changes in soluble protein content during forelimb regeneration in the adult newt,Notophthalmus viridescens. J Exp Zool 190:317–328

  6. Foret JE (1973) Stimulation and retardation of limb regeneration by adenosine-3′,5′-monophosphate and relate compunds. Oncology 27:153–159

  7. Foret JE, Babich, GL (1973) Effects of dibutyryl cyclic AMP and related compounds on newt limb regeneration blastemas in vitro. I.3H-thymidine incorporation. Oncology 28:83–88

  8. Iten LE, Bryant SV (1973) Forelimb regeneration from different levels of amputation in the newt,Notophthalmus viridescens: length, rate, stage. Wilhelm Roux's Arch 173:263–282

  9. Jabaily J, Rall TW, Singer M (1975) Assay of cyclic 3′,5′-monophosphate in the regenerating forelimb of the newt,Triturus. J Morphol 147:373–384

  10. Kosher RA, Savage MP (1980) Studies on the possible role of cyclic AMP in limb morphogenesis and differentiation. J Embryol Exp Morphol 56:91–105

  11. Krebs E (1972) Protein kinases. Curr Topics Cell Regulat 5:99–133

  12. Lee P, Jungmann R (1975) Ontogeny of cyclic AMP-dependent protein phosphokinase during hepatic development of the rat. Biochim Biophys Acta 399:265–276

  13. Liversage RA, Rathbone MP, McLaughlin HMG (1977) Changes in cyclic GMP levels during forelimb regeneration in adultNotophthalmus viridescens. J Exp Zool 200:169–175

  14. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurements with the folin-phenol reagent. J Biol Chem 193:265–275

  15. Rathbone MP, Petri J, Choo AF, Logan DM, Carlone RL, Foret JE (1980) Noradrenaline and cyclic AMP-independent growth stimulation in newt limb blastemata. Nature 283:387–388

  16. Robison GA, Butcher RW, Sutherland EW (1971) Cyclic AMP. Academic Press, New York

  17. Sicard RE (1975a) Cyclic AMP and neuroendocrine influence upon forelimb regeneration in the adult newt,Notophthalmus viridescens. Oncology 32:190–195

  18. Sicard, RE (1975b) The effects of hypophysectomy upon the endogenous levels of cyclic AMP during forelimb regeneration of adult newts (Notophthalmus viridescens). Wilhelm Roux's Arch 177:159–162

  19. Solursh M, Ahrens PB, Reiter RS (1978) A tissue culture analysis of the steps in limb chondrogenesis. In Vitro 14:51–61

  20. Taban C, Cathieni M, Schorderet M (1978) Cyclic AMP and noradrenaline sensitivity fluctuations in regenerating newt tissue. Nature 271:470–472

  21. Traugh JA, Ashby CD, Walsh DA (1974) Criteria for the classification of protein kinases. In: Colowick SP, Kaplan NO (eds) Methods in Enzymology vol XXXVIII, Part C. Academic Press, New York, pp 290–299

Download references

Author information

Correspondence to Raymond E. Sicard.

Additional information

Portions of this work constitute part of the thesis submitted by T.M.L. in partial fulfillment of the requirements for the M.S. degree in Biology at Boston College

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Laz, T.M., Sicard, R.E. Cyclic nucleotide metabolism during amphibian forelimb regeneration. Wilhelm Roux' Archiv 191, 163–168 (1982). https://doi.org/10.1007/BF00848331

Download citation

Key words

  • Notophthalmus viridescens
  • Regeneration
  • Cyclic nucleotides
  • cAMP
  • Protein kinases