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Lysosomal acid phosphatase mediates dedifferentiation in the regenerating salamander limb

  • Ju, Bong-Gun (Department of Life Science, Sogang University) ;
  • Kim, Won-Sun (Department of Life Science, Sogang University)
  • Received : 2010.03.07
  • Published : 2010.06.30

Abstract

In this study, monoclonal antibodies against lysosomal acid phosphatase (LAP) of a salamander, Hynobius leechii, were used to determine the spatial and temporal expression of the LAP in the regenerating limbs. The Western blot and immunohistochemical analysis in the limb regeneration revealed that LAP was highly expressed at the dedifferentiation stage, especially in the wound epidermis and dedifferentiating limb tissues such as muscle and cartilage. With RA treatment, the LAP expression became upregulated in terms of both level and duration in the wound epidermis, blastemal cell and dedifferentiating limb tissues. In addition, in situ activity staining of LAP showed a similar result to that of immunohistochemistry. Thus, the activity profile of LAP activity coincides well with the expression profile of LAP during the dedifferentiation period. Furthermore, to examine the effects of lysosomal enzymes including LAP on salamander limb regeneration, lysosome extract was microinjected into limb regenerates. Interestingly, when the lysosome extract was microinjected into limb regenerates with a low dose of RA($50\;{\mu}g/g$ body wt.), skeletal pattern duplication occurred frequently in the proximodistal and transverse axes. Therefore, lysosomal enzymes might cause the regenerative environment and RA plays dual roles in the modification of positional value as well as evocation of extensive dedifferentiation for pattern duplication. In conclusion, these results support the hypothesis that dedifferentiation is a crucial event in the process of limb regeneration and RA-evoked pattern duplication, and lysosomal enzymes may play important role(s) in this process.

Acknowledgement

Supported by : National Research Foundation of Korea (NRF)

References

  1. Alibardi L. 1998. Presence of acid phosphatase in the epidermis of the regenerating tail of the lizard (Podarcis muralis) and its possible role in the process of shedding and keratinization. J Zool. 246:379-390. https://doi.org/10.1111/j.1469-7998.1998.tb00170.x
  2. Bohlen P, Stein S, Dairman W, Udenfriend S. 1973. Fluorometric assay of proteins in the nanogram range. Arch Biochem Biophys. 155:213-220. https://doi.org/10.1016/S0003-9861(73)80023-2
  3. Boivin P, Galand C. 1986. The human red cell acid phosphatase is a phosphotyrosine protein phosphatase which dephosphorylates the membrane protein band 3. Biochem Biophys Res Commun. 134:557-564. https://doi.org/10.1016/S0006-291X(86)80456-9
  4. Bryant SV, Iten LE. 1974. The regulative ability of the limb regeneration blastema of Triturus viridescens. Roux's Arch Dev Biol. 174:90-101. https://doi.org/10.1007/BF00577059
  5. Burstone MS. 1958. Histochemical demonstration of acid phosphatase with naphthol AS-phosphates. J Natl Cancer Inst. 21 :523-539.
  6. Carlson BM. 1974. Factors controlling the initiation and cessation of early events in regenerative process. In: Sherbet GV, editor. Neoplasia and cell differentiation. Basel: Karger. p. 60-105.
  7. Chernoff J, Li HC. 1985. A major phosphotyrosyl-protein phosphatase from bovine heart is associated with a low-molecular weight acid phosphatase. Arch Biochem Biophys. 240:135-145. https://doi.org/10.1016/0003-9861(85)90016-5
  8. Coward SJ, Bennett CE, Hazlehurst BL. 1973. Lysosomes and lysosomal enzyme activity in the regenerating planarian: evidence in support of dedifferentiation. J Exp Zool. 189:133-146.
  9. Deck JD, Dent JN. 1970. The effects of infused material upon regeneration of newt limbs. Blastemal extracts and alkaline phosphatase in irradiated limb stumps. Anat Rec. 168:525-535. https://doi.org/10.1002/ar.1091680405
  10. De Duve C. 1983. Lysosomes revisited. Eur J Biochem. 137:391-397. https://doi.org/10.1111/j.1432-1033.1983.tb07841.x
  11. Dukiet B, Niwelinski J. 1960. Localization of $\beta$-glucuronidase in the regenerating forelimb of adult, Triturus alpestris. Folia Biol. 8:291-298.
  12. Globus M, Vethamany-Globus S. 1976. An in vitro analogue of early chick limb bud outgrowth. Differentiation. 6:91-96. https://doi.org/10.1111/j.1432-0436.1976.tb01474.x
  13. Grillo JC, Lapiere CM, Dresden MH, Gross J. 1968. Collagenolytic activity in regenerating forelimbs of the adult newt (Triturus viridescens). Dev Biol. 17:571-583. https://doi.org/10.1016/0012-1606(68)90006-7
  14. Holtzman E. 1989. Lysosomes. New York: Plenum Press. p.243-316.
  15. Janska H, Kubicz A, Szalewicz A, Harazna J. 1988. The high molecular weight and the low molecular weight acid phosphatases of the frog liver and their phosphotyrosine activity. Comp Biochem Physiol. 90B:173-178.
  16. Jones HE, Bowen ID. 1993. Acid phosphatase activity in the larval salivary glands of developing Drosophila melanogaster. Cell Biol Int. 17:305-315. https://doi.org/10.1006/cbir.1993.1066
  17. Ju BG, Kim WS. 1994. Pattern duplication by retinoic acid treatment 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
  18. Ju BG, Kim WS. 2004. Cloning of a cDNA Encoding cathepsin D from salamander, Hynobius leechii, and its expression in the limb regenerates. DNA Seq. 11:21-28.
  19. Ju BG, Park SY, Choi EY, Kim WS. 1996. Lysosomal acid phosphatase in regenerating salamander limbs studied with monoclonal antibodies. Korean J Zool. 39:426-436.
  20. Kim WS, Stocum DL. 1986. Effects of retinoic acid on regenerating normal and double half limbs of axolotls, Roux's Arch Dev Biol. 195:243-251. https://doi.org/10.1007/BF02438957
  21. Kumar S, Khanduja KL, Verma N, Verma SC, Avti PK, Pathak CM. 2008. ATRA promotes alpha tocopherol succinate-induced apoptosis in freshly isolated leukemic cells from chronic myeloid leukemic patients. Mol Cell Biochem. 307:109-119. https://doi.org/10.1007/s11010-007-9590-7
  22. Ling BY, Lyerla TA. 1976. Acid phosphatase activity in the development of the cement gland in Xenopus laevis. J Exp Zool. 195:191-198. https://doi.org/10.1002/jez.1401950203
  23. Lippincott-Schwart J, Fambrough DM. 1986. Lysosomal membrane dynamics: structure and itraorganellar movement of a major lysosomal membrane glycoprotein. J Cell Biol. 102:1593-1605. https://doi.org/10.1083/jcb.102.5.1593
  24. Mahapatra PK, Mohanty-Hejmadi P, Chainy GB. 2001. Changes in oxidative stress parameters and acid phosphatase activity in the pre-regressing and regressing tail of Indian jumping frog Polypedates maculatus (Anura, Rhacophoridae). Comp Biochem Physiol C Toxicol Pharmacol. 130:281-288. https://doi.org/10.1016/S1532-0456(01)00238-1
  25. Milaire J. 1997. A comparative review of the site of histoenzymatic dephosphorylating activities and of gene expression in developing limb buds of mammalian and avian species. Eur J Morpho. 35:143-169. https://doi.org/10.1076/ejom.35.3.143.13071
  26. Miller NR. Wolfe HJ. 1968. The nature and localization of acid phosphatase during the early phases of urodele limb regeneration. Dev Biol. 17:447-481. https://doi.org/10.1016/0012-1606(68)90074-2
  27. Neufeld DA. 1980. Partial blastema formation after amputation in adult mice. J Exp Zool. 212:31-36. https://doi.org/10.1002/jez.1402120105
  28. Oreffo ROC, Tetti A, Triffitt JT, Francis MJO, Carano A, Zallone AZ. 1988. Effect of vitamin A on bone resorption: evidence for direct stimulation of isolated chicken osteoclasts by retinol and retinoic acid. J Bone Mineral Res. 3:203-210.
  29. Pavlovic B, Brunati AM, Barbaric S, Pinna RA. 1985. Repressible acid phosphatase from yeast efficiently dephosphorylates in vivo some phosphorylated proteins and peptides. Biochem Biophys Res Commun. 129:350-357. https://doi.org/10.1016/0006-291X(85)90158-5
  30. Rabossi A, Stoka V, Puizdar V, Turk V, Quesada-Allue LA. 2004. Novel aspartyl proteinase associated to fat body histolysis during Ceratitis capitata early metamorphosis. Arch Insect Biochem Physiol. 57:51-67. https://doi.org/10.1002/arch.20011
  31. Robinson H. 1970. Acid phosphatase in the tail of Xenopus laevis during development and metamorphosis. J Exp Zool. 173:215-224. https://doi.org/10.1002/jez.1401730209
  32. Robinson H. 1972. An electrophoretic and biochemical analysis of acid phosphatase in the tail of Xenopus laevis during development and metamorphosis. J Exp Zool. 180:127-140. https://doi.org/10.1002/jez.1401800111
  33. Roles OA, Anderson OR, Lin NST, Shah DO, Trout ME. 1969. Vitamin A and membranes. Amer J Clin Nutr. 22:1020-1023. https://doi.org/10.1093/ajcn/22.8.1020
  34. Schmidt AJ. 1968. Cellular biology of vertebrate regeneration and repair. Chicago: University of Chicago Press. p. 17-117.
  35. Schmidt AJ, Norman W. 1965. The distribution of esterase in the regenerating and non-regenerating tissues or the adult newt forelimb. Anat Rec. 151:474-475.
  36. Schimidt AJ, Weidman T. 1964. Dehydrogenase and aldolase in the regenerating forelimb of the adult newt, Diemictylus viridescens. J Exp Zool. 155:303-316. https://doi.org/10.1002/jez.1401550304
  37. Stocum DL. 1979. Stages of forelimb regeneration in Ambystoma maculatum. J Exp Zool. 209:395-416. https://doi.org/10.1002/jez.1402090306
  38. Stocum DL. 1995. Wound repair, regeneration and artificial tissues. Austin: R.G. Landes. p. 51-223.
  39. Symons L, Jonas AJ. 1987. Isolation of highly purified rat liver lysosomal membranes using two Percoll gradients. Anal Biochem. 164:382-390.
  40. Takei Y, Higashira H, Yamamoto T, Hayashi K. 1997. Mitogenic activity toward human breast cancer cell line MCF-7 of two bFGFs purified from sera of breast cancer patients: co-operative role of cathepsin D. Breast Cancer Res Treat. 43:53-63. https://doi.org/10.1023/A:1005749925296
  41. Thoms SD, Stocum DL. 1984. Retinoic acid-induced pattern duplication in regenerating urodele limbs. Dev Biol. 103:319-328. https://doi.org/10.1016/0012-1606(84)90320-8
  42. Wang CC, Straight S, Hill DL. 1976. Destabilization of mouse liver lysosomes by vitamin A compounds and analogues. Biochem Pharmacol. 25:471-475. https://doi.org/10.1016/0006-2952(76)90351-8
  43. Weber R. 1957. On the biological function of cathepsin in tail tissues of Xenopus larvae. Exp Cell Res. 15:1-20.
  44. Weber R. 1963. Behavior and properties of acid hydrolases in the regressing tails of tadpoles during spontaneous and induced metamorphosis in vitro. In: de Renck AVR, Cameron MP, editors. Lysosomes. Boston: Little, Brown. p.282-310.
  45. Weiss C, Rosenbaum RM. 1968. Histochemical studies on cell death and histolysis during regeneration. I. Distribution of acid phosphomonoesterase activity in the normal, the regenerating and the resorbing forelimb of the larval spotted salamander, Amblystoma maculatum. J Morph. 122:203-230.