Analysis of TIMP-2 and Vimentin Protein Expression and Epigenetic Reprogramming in Cloned Bovine Placentae

  • Kim, Hong-Rye (Research Center for Transgenic Cloned Pigs, Chungnam National University) ;
  • Han, Rong-Xun (Research Center for Transgenic Cloned Pigs, Chungnam National University) ;
  • Lee, Hye-Ran (Research Center for Transgenic Cloned Pigs, Chungnam National University) ;
  • Yoon, Jong-Taek (Genetic Engineering Institute, Hankyong National University) ;
  • Cheong, Hee-Tae (School of Veterinary Medicine, Kangwon National University) ;
  • Park, Chang-Sik (Research Center for Transgenic Cloned Pigs, Chungnam National University) ;
  • Jin, Dong-Il (Research Center for Transgenic Cloned Pigs, Chungnam National University)
  • Published : 2007.06.30

Abstract

The objective of this study was to analyzed pattern of proteins expression abnormally in cloned bovine placenta. TIMP-2 protein whose function is related to extracellular matrix degradation and tissue remodeling processes was one of differentially up-regulated proteins in SCNT placenta. And one of down-regulated protein in SCNT placenta was identified as vimentin protein that is presumed to stabilize the architecture of the cytoplasm. The expression patterns of these proteins were validated by Western blotting. To evaluate how regulatory loci. of TIMP-2 and vimentin genes was programmed reprogramming in cloned placenta. the status of DNA methylation in the promoter region of TIMP-2 and vimentin genes was analyzed by sodium Bisulfite mapping. The DNA methylation results showed that there was not difference in methylation pattern of TIMP-2 and vimentin loci between cloned and normal placenta. Histone H3 acetylation state of the nucleosome was analyzed in the cloned placental and normal placenta by Western blotting. A small portion of the protein lysates were subjected to Western blotting with the antibodies against anti acetyl-Histone H3. Overall histone H3 acetylation state of SCNT placenta was significantly higher than those of normal placenta cells. It is postulated that cloned placenta at the end of gestation seems to be unusual in function and morphology of placenta via improper expression of TIMP-2 and vimentin by abnormal acetylation states of cloned genome.

Keywords

References

  1. Byun Y, Chen F, Chang R, Trivedi M, Green KJ, Cryns VL (2001): Caspase cleavage of vimentin disrupts intermediate filaments and promotes apoptosis. Cell Death Differ 8:443-450 https://doi.org/10.1038/sj.cdd.4400840
  2. Cezar GG, Bartolomei MS, Forsberg EJ, First NL, Bishop MD, Eilertsen KJ (2003): Genome-wide epigenetic alterations in cloned bovine fetuses. Biol Reprod 68:1009-1014 https://doi.org/10.1095/biolreprod.102.010181
  3. Fry CJ, Peterson CL (2001): Chromatin remodeling enzymes: who's on first? Curr Biol 11:185-197 https://doi.org/10.1016/S0960-9822(01)00044-6
  4. Han YM, Kang YK, Koo DB, Lee KK (2003): Nuclear reprogramming of cloned embryos produced in vitro. Theriogenology 59:33-44 https://doi.org/10.1016/S0093-691X(02)01271-2
  5. Hashimoto M, Inoue S, Ogawa S, Conrad C, Muramatsu M, Shackelford D, Masliah E (1998): Rapid fragmentation of vimentin in human skin fibroblasts exposed to tamoxifen: a possible involvement of caspase-3. Biochem Biophys Res Commu 247:401-406 https://doi.org/10.1006/bbrc.1998.8799
  6. Hill JR, Burghardt RC, Jones K, Long CR, Looney CR, Shin T, Spencer TE, Thompson JA, Winger QA, Westhusin ME (2000): Evidence for placental abnormality as the major cause of mortality in first-trimester somatic cell cloned bovine fetuses. Biol Reprod 63:1787-1794 https://doi.org/10.1095/biolreprod63.6.1787
  7. Humpherys D, Eggan K, Akutsu H, Friedman A, Hochedlinger K, Yanagimachi R, Lander ES, Golub TR, Jaenisch R (2002): Abnormal gene expression in cloned mice derived from embryonic stem cell and cumulus cell nuclei. Proc Natl Acad Sci USA 99: 12889-12894
  8. Jenuwein T, Allis CD (2001): Translating the histone code. Science 293:1074-1080 https://doi.org/10.1126/science.1063127
  9. Jones PA, Takai D (2001): The role of DNA methylation in mammalian epigenetics. Science 293: 1068-1070 https://doi.org/10.1126/science.1063852
  10. Kang YK, Park JS, Koo DB, Choi YH, Kim SU, Lee KK, Han YM (2002): Limited demethylation leaves mosaic-type methylation states in cloned bovine preimplantation embryos. EMBO 21:1092-1100 https://doi.org/10.1093/emboj/21.5.1092
  11. Kim HR, Kang JK, Yoon JT, Seong HH, Jung JK, Lee HM, Sik Park C, Jin DI (2005): Protein profiles of bovine placenta derived from somatic cell nuclear transfer. Proteomics 5:4264-4273 https://doi.org/10.1002/pmic.200401297
  12. McGraw S, Robert C, Massicotte L, Sirard MA (2003): Quantification of histone acetyltransferase and histone deacetylase transcripts during early bovine embryo development. Biol Reprod 68:383-389 https://doi.org/10.1095/biolreprod.102.005991
  13. Morishima N (1999): Changes in nuclear morphology during apoptosis correlate with vimentin cleavage by different caspases located either upstream or downstream of Bcl-2 action. Genes Cells 4:401-414 https://doi.org/10.1046/j.1365-2443.1999.00270.x
  14. Oswald J, Engemann S, Lane N, Mayer W, Olek A, Fundele R, Dean W, Reik W, Walter J (2000): Active demethylation of the paternal genome in the mouse zygote. Curr Biol 10:475-478 https://doi.org/10.1016/S0960-9822(00)00448-6
  15. Prasad SC, Thraves PJ, Kuettel MR, Srinivasarao GY, Dritschilo A, Soldatenkov VA (1998): Apoptosis-associated proteolysis of vimentin in human prostate epithelial tumor cells. Biochem Biophys Res Commun 249:332-338 https://doi.org/10.1006/bbrc.1998.9137
  16. Reik W, Santos F, Dean W (2003): Mammalian epigenomics: reprogramming the genome for development and therapy. Theriogenology 59:21-32 https://doi.org/10.1016/S0093-691X(02)01269-4
  17. Suemizu H, Aiba K, Yoshikawa T, Sharov AA, Shimozawa N, Tamaoki N, Ko MS (2003): Expression profiling of placentomegaly associated with nuclear transplantation of mouse ES cells. Dev Biol 253:36-53 https://doi.org/10.1006/dbio.2002.0870
  18. Van Engeland M, Kuijpers HJ, Ramaekers FC, Reutelingsperger CP, Schutte B (1997): Plasma membrane alterations and cytoskeletal changes in apoptosis. Exp Cell Res 235:421-430 https://doi.org/10.1006/excr.1997.3738
  19. Wakayama T, Yanagimachi R (2001): Mouse cloning with nuclear donor cells of different age and type. Mol. Reprod 58:376-383 https://doi.org/10.1002/1098-2795(20010401)58:4<376::AID-MRD4>3.0.CO;2-L
  20. Walter I, Boos A (2001): Matrix metalloproteinases (MMP-2 and MMP-9) and tissue inhibitor-2 of matrix metalloproteinases (TIMP-2) in the placenta and interplacental uterine wall in normal cows and in cattle with retention of fetal membranes. Placenta 22:473-483 https://doi.org/10.1053/plac.2001.0633
  21. Williams WF, Margolis MJ, Manspeaker J, Douglass LW, Davidson JP (1987): Peripartum changes in the bovine placenta related to fetal membrane retention. Theriogenology 28:213-223 https://doi.org/10.1016/0093-691X(87)90268-8
  22. Young LE, Sinclair KD, Wilmut I (1998): Large offspring syndrome in cattle and sheep. Rev Reprod 3:155-163 https://doi.org/10.1530/ror.0.0030155