Animal cells and systems

The Korean Society for Integrative Biology (한국통합생물학회)
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Protein Synthesis Pattern Analysis in the Regenerating Salamander Limb

  • Ju, Bong-Gun (Department of Life Science, College of Sciences, Sogang University) ;
  • Kim, Won-Sun (Department of Life Science, College of Sciences, Sogang University)
  • Published : 2000.06.01
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Abstract

Retinoic acid (RA) evokes pattern duplication in the regenerating salamander limb. Interestingly, it also enhances dedifferentiation in the regenerate by the morphological, histological and biochemical criteria. To examine whether there is any correlation between the RA-evoked pattern duplication and de novo protein synthetic profile in the regenerating salamander limb, especially during dedifferentiation, we analyzed stage-specific protein synthesis pattern in the normal and RA-treated regenerating limbs by metabolic labeling followed by two-dimensional gel electrophoresis. In the regenerating limbs without RA treatment, a few hundred kinds of proteins were found to be synthesized at the stage of wound healing and the total number of protein synthesized increased greatly as regeneration proceeded. The same trend was also observed in the RA-treated regenerating limbs. Interestingly, some protein spots were noted to be either newly synthesized or highly expressed by the RA treatment especially at the stage of dedifferentiation. The results shows that the enhancement of dedifferentiation state after the RA treatment correlates well with the protein synthesis profile, and suggest that those proteins are important for the RA-evoked pattern duplication in the regenerating limbs of salamander.

Keywords

References

  1. Bodemer CW; and Everett NB (1959) Localization of newly synthesizes proteins in regenerating newt limb as determined by redioautogeraphic loca;ization of injected methionine-$^{35}S$. Dev Biol 1: 327-342 https://doi.org/10.1016/0012-1606(59)90032-6
  2. Bonner WM and Laskey RA (1974) A film detection method for tritium-labeled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem 86: 83-88 https://doi.org/10.1111/j.1432-1033.1974.tb03599.x
  3. Carlone RL, Boulianne RP, Vijh KM, Kahn H, and Fraser GAP (1993) Retinoic acid stimulates the synthesis of a novel heat shock protein in the regenerating forelimb of the newt. Biochem Cell Biol 71: 43-50
  4. Carlson BM (1967) The histolgy of inhibition of limb regeneration in the newt, Triturus, by actinomycin D. J Morphol 122: 246-264 https://doi.org/10.1002/jmor.1051220306
  5. Dukiet B; and Niwelinski J (1960) Localization of ${\beta}$-giucuronidase in the regenerating forelimb of adult, Triturus alpestris. Folia Biol 8: 291-298
  6. Giguere V, Ong ES, Evans RM, and Tabin CJ (1989) Spatial and temporal expression of the retinoic acid receptor in the regenerating amphibian limb. Nature 337: 566-569 https://doi.org/10.1038/337566a0
  7. Grillo JC, Lapiere CM, Dresden MH, and Gross J (1968) Collagenolytic activity in regenerating forelimbs of the newt (Triturus viridescens). Dev Biol 17: 571-583 https://doi.org/10.1016/0012-1606(68)90006-7
  8. Ju BG and Kim WS (1994) Pattern duplication asic treatmwnt in the regenerating limbs of korean salamander larvae,Hynobius Leechii, correlates well with the extent of dedifferentiation. Dev Dyn 199: 253-267 https://doi.org/10.1002/aja.1001990402
  9. Ju BG and Kim WS (1998) Upregulation of cathepsin D expression in the dedifferentiating salamander limb regenerates and enhancement of its expression by retinoic acid. Wound Rep Reg 6: 349-357 https://doi.org/10.1046/j.1524-475X.1998.60410.x
  10. Kim WS and Stocum DL (1986) Retinoic asid modifies positional memory in the anteroposterior axis of regenerating axolotl limbs. Dev Biol 114: 170-179 https://doi.org/10.1016/0012-1606(86)90393-3
  11. Lee EH and Kim WS (1996) Dedifferentiation state specific increase of trypsin- and chymotrypsin-like protease activities during urodele limb regeneration and their enhancement by retinoic acid treatment. Korean J Zool 39: 65-74
  12. Ludolph DC, Cameron JA, and Stocum DL (1990) The effect of retinoic acid on the positional memory in the dorsoventral axis of the regenerating axolotl limbs. Dev Biol 140: 41-52 https://doi.org/10.1016/0012-1606(90)90051-J
  13. Maden M (1982) Vitamin A and pattern formation in the regenerating limb. Nature 295: 72-75 https://doi.org/10.1038/295672a0
  14. Maden M, Keeble S, and Cox RA (1985) The characteristics of local application of retinoic acid to the regenerating axolotl limb. Roux's Arch Dev Biol 194: 228-235 https://doi.org/10.1007/BF00848251
  15. Mohun TJ, Tilly R, and Slack JMW (1980) Cell commitment and gene expression in the axolotl embryo. Cell 22: 9-15 https://doi.org/10.1016/0092-8674(80)90149-X
  16. Niazi IA, Pescitelli MJ, and Stocum DL (1985) Stage-dependent effects of retinoic acid on regenerating urodele limbs. Roux's Arch. Dev Biol 194: 355-363 https://doi.org/10.1007/BF00877373
  17. Niazi IA and Saxena S (1978) Abnormal hind limb regeneration in tadpoles of the toad,Bufo andedersoni, exposed to excess vitamin A. Folia Biol 26: 3-11
  18. O'Farrell PH (1975) High resolution two-dimensional electrophoresis of proteins. J Biol Chem 250: 4007-4021
  19. Park IS and Kim WS (1998) Modulation of gelatinase activity correlates with the dedifferentiation profile of regenerating salamander Limb. Mol Cells 9: 119-126
  20. Ragsdale CW, Gates PB, Hill DS, and Brockes JP (1992) Delta retinoic acid receptor isoform ${\\delta}_1$is distinguisfed by its N-terminal sequence and abundance in the limb regeneration blastema. Mech Dev 40: 99-112 https://doi.org/10.1016/0925-4773(93)90091-B
  21. Ragsdale CW, Petikovich M, Gate PB, Chambon P, and Brocks JP (1989) Identification of novel retinoic acid receptor in regenerative tissues of the newt. Nature 341: 654-657 https://doi.org/10.1038/341654a0
  22. Ramagli LS, Rodriguez LV (1985) Quantitation of microgram amounts of protein in two-dimensional polyasrylamidegel electrophoresis sample buffer. Electrophoresis 6: 559-563 https://doi.org/10.1002/elps.1150061109
  23. Rose SM (1970) Regeneration. Appletion-Century-Crofts, New York, pp 39-78
  24. Scadding SR and Maden M (1986) Comparison of the effects of vitamin A on limb development and regeneration in the axolotl, Ambystoma mexicanum. J Embryol Exp Morphol 91: 19-34
  25. Schmidt AJ (1968) Cellular Biology of Vertebrate Regeneration and Repair. The University of Chicago Press, Chicago, pp 17-117
  26. Schmidt AJ and Norman W (1965) The distribution of esterase in hte regenerating and non-regenerating tissues or the adult newt forelimb. Anat Rec 151: 474-475
  27. Schmidt AJ;Weidman T (1964) Dehydrogenase and aldolase in the regenerating forelimb of the adult newt, Diemistylus viridescens. J Exp Zool 155: 303-316 https://doi.org/10.1002/jez.1401550304
  28. Slack JMW (1982) Protein synthesis during limb regeneration in the axolotl. J Embryol Exp Morphol 70: 241-260
  29. Slattery GG and Schmidt AJ (1975) The effect of dimethyl suifoxide on forelimb regeneration of the adult newt, Triturus viridescens. Ann N Y Acad Sci 243: 257-268 https://doi.org/10.1111/j.1749-6632.1975.tb25364.x
  30. Stocum DL (1995) Wound Repair Regeneration and Artificial Tissues. R. G. Landes Company, Austin
  31. Thoms SD and Stocum DL (1984) Retinoic acid-induced pattern duplication in regenerating urdele limbs. Dev Biol 103: 319-328 https://doi.org/10.1016/0012-1606(84)90320-8