Expression and DNA Methylation Change of Oct-4 in Cloned Bovine Blastocysts

체세포복제 소 배반포의 Oct-4 발현과 DNA 메틸화 변화

  • 차병현 (농촌진흥청 축산과학원 응용생명공학과) ;
  • 최정상 (농촌진흥청 축산과학원 응용생명공학과) ;
  • 황성수 (농촌진흥청 축산과학원 응용생명공학과) ;
  • 정학재 (농촌진흥청 축산과학원 응용생명공학과) ;
  • 임기순 (농촌진흥청 축산과학원 응용생명공학과) ;
  • 양병철 (농촌진흥청 축산과학원 응용생명공학과) ;
  • 김명직 (농촌진흥청 축산과학원 응용생명공학과) ;
  • 조재현 (경상대학교 수의학과) ;
  • 성환후 (농촌진흥청 축산과학원 응용생명공학과) ;
  • 고응규 (농촌진흥청 축산과학원 응용생명공학과)
  • Published : 2008.09.30

Abstract

DNA methylation is one of the reasons for poor survival of clone animals. The OCT-4 gene is essential for maintaining pluripotency of embryonic stem (ES) cells and early embryos. We previously reported that the 5'-promoter region of Oct-4 gene was a target of DNA methylation and the methylation status was changed variously during embryonic development in bovine. The study conducted to examine the expression and methylation pattern of tissue-dependent differentially methylated region (T-DMR) of Oct-4 gene in bovine somatic cell nuclear transfer (SCNT) and in vitro fertilization (IVF) blastocysts. The Oct-4 gene expression was evaluated by RT-PCR and fluorescence immunocytochemistry. The methylation pattern of T-DMR was analyzed using restriction mapping and bisulfite sequencing methods. The Oct-4 transcripts were highly expressed in IVF, while they were not expressed in SCNT. The Oct-4 protein was not detected or expressed at very low level in SCNT, the intensity of Oct-4 protein, however, was strong in IVF. On the other hand, the T-DMR of Oct-4 gene was hypermethylated in SCNT compared to that of IVF. These results suggested that expression and the failure of demethylation of Oct-4 gene was closely associated with incomplete development of SCNT embryos.

Keywords

References

  1. Beaujean N, Taylor JE, McGarry M, Gardner JO, Wilmut I, Loi P, Ptak G, Galli C, Lazzari G, Bird A, Young LE and Meehan RR. 2004. The effect of interspecific oocytes on demethylation of sperm DNA. Proc. Natl. Acad. Sci. USA 101:7636-7640
  2. Boiani M, Eckardt S, Scholer HR and McLaughlin KJ. 2002. Oct-4 distribution and level in mouse clones: consequences for pluripotency. Genes. Dev. 16:1209-1219 https://doi.org/10.1101/gad.966002
  3. Bourc'his D, Le Bourhis D, Patin D, Niveleau A, Comizzoli P, Renard JP and Viegas-Pequignot E. 2001. Delayed and incomplete reprogramming of chromosome methylation patterns in bovine cloned embryos. Curr. Biol. 11:1542-1546 https://doi.org/10.1016/S0960-9822(01)00480-8
  4. Hattori N, Nishino K, Ko YG, Hattori N, Ohgane J, Tanaka S and Shiota K. 2004. Epigenetic control of mouse Oct-4 gene expression in embryonic stem cells and trophoblast stem cells. J. Biol. Chem. 279:17063-17069 https://doi.org/10.1074/jbc.M309002200
  5. Hill JR, Winger QA, Long CR, Looney CR, Thompson JA and Westhusin ME. 2000. Development rates of male bovine nuclear transfer embryos derived from adult and fetal cells. Biol. Reprod. 62:1135-1140 https://doi.org/10.1095/biolreprod62.5.1135
  6. Im GS, Yang BS, Yang BC, Chang WK, Yi YJ and Park CS. 2001. Effect of cell cycle stage on the development of embryos produced by cumulus cell nuclear transfer in Hanwoo (Korea cattle). Asian-Ausu. J. Anim. Sci. 14:759-764 https://doi.org/10.5713/ajas.2001.759
  7. Kang YK, Koo DB, Park JS, Choi YH, Chung AS, Lee KK and Han YM. 2001. Aberrant methylation of donor genome in cloned bovine embryos. Nat. Genet. 28:173-177 https://doi.org/10.1038/88903
  8. Kato Y, Tani T, Sotomaru Y, Kurokawa K, Kato J, Doguchi H, Yasue H and Tsunoda Y. 1998. Eight calves cloned from somatic cells of a single adult. Science 282:2095-2098 https://doi.org/10.1126/science.282.5396.2095
  9. Kim JM, Ko YG, Seong HH, Chung HJ, Chang WK and Kim NH (2007): Alteration of DNA methylation in Oct-4 gene in mouse preimplantation embryos by the interference RNA. Reprod. Dev. Biol. 31:21-28
  10. Kirchhof N, Carnwath JW, Lemme E, Anastassiadis K, Scholer H and Niemann H. 2000. Expression pattern of Oct-4 in preimplantation embryos of different species. Biol. Reprod. 63:1698-1705 https://doi.org/10.1095/biolreprod63.6.1698
  11. Ko YG, Nishino K, Hattori N, Arai Y, Tanaka S and Shiota K. 2005. Stage-by-stage change in DNA methylation status of Dnmt1 locus during mouse early development. J. Biol. Chem. 280:9627-9634 https://doi.org/10.1074/jbc.M413822200
  12. Kremenskoy M, Kremenska Y, Suzuki M, Imai K, Takahashi S, Hashizume K, Yagi S and Shiota K. 2006. Epigenetic characterization of the CpG islands of bovine Leptin and POU5F1 genes in cloned bovine fetuses. J. Reprod. Dev. 52: 277-285 https://doi.org/10.1262/jrd.17100
  13. Kubota C, Yang X, Dinnyes A, Todoroki J, Yamakuchi H, Mizoshita K, Inohae S and Tabara N. 1998. In vitro and in vivo survival of frozen-thawed bovine oocytes after IVF, nuclear transfer, and parthenogenetic activation. Mol. Reprod. Dev. 51:281-286 https://doi.org/10.1002/(SICI)1098-2795(199811)51:3<281::AID-MRD7>3.0.CO;2-L
  14. Mizoshita K, Watanabe T, Hayashi H, Kubota C, Yamakuchi H, Todoroki J and Sugimoto Y. 2004. Quantitative trait loci analysis for growth and carcass traits in a half-sib family of purebred Japanese Black (Wagyu) cattle. J. Anim. Sci. 82:3415-3420
  15. Nordhoff V, Hubner K, Bauer A, Orlova I, Malapetsa A and Scholer HR. 2001. Comparative analysis of human, bovine, and murine Oct-4 upstream promoter sequences. Mamm. Genome 12:309-317 https://doi.org/10.1007/s003350010279
  16. Ohgane J, Wakayama T, Kogo Y, Senda S, Hattori N, Tanaka S, Yanagimachi R and Shiota K. 2001. DNA methylation variation in cloned mice. Genesis 30:45-50 https://doi.org/10.1002/gene.1031
  17. Ohgane J, Wakayama T, Senda S, Yamazaki Y, Inoue K, Ogura A, Marh J, Tanaka S, Yanagimachi R and Shiota K. 2004. The Sall3 locus is an epigenetic hotspot of aberrant DNA methylation associated with placentomegaly of cloned mice. Genes. Cells 9:253-260 https://doi.org/10.1111/j.1356-9597.2004.00720.x
  18. Okamoto K, Okazawa H, Okuda A, Sakai M, Muramatsu M and Hamada H. 1990. A novel octamer binding transcription factor is differentially expressed in mouse embryonic cells. Cell 60:461-472 https://doi.org/10.1016/0092-8674(90)90597-8
  19. Polejaeva IA, Chen SH, Vaught TD, Page RL, Mullins J, Ball S, Dai Y, Boone J, Walker S, Ayares DL, Colman A and Campbell KH. 2000. Cloned pigs produced by nuclear transfer from adult somatic cells. Nature 407:86-90 https://doi.org/10.1038/35024082
  20. Renard JP, Chastant S, Chesne P, Richard C, Marchal J, Cordonnier N, Chavatte P and Vignon X. 1999. Lymphoid hypoplasia and somatic cloning. Lancet 353:1489-1491 https://doi.org/10.1016/S0140-6736(98)12173-6
  21. Rhind SM, Taylor JE, De Sousa PA, King TJ, McGarry M and Wilmut I. 2003. Human cloning: can it be made safe? Nat. Rev. Genet. 4:855-864
  22. Rosner MH, Vigano MA, Ozato K, Timmons PM, Poirier F, Rigby PW and Staudt LM. 1990. A POU-domain transcription factor in early stem cells and germ cells of the mammalian embryo. Nature 345:686-692 https://doi.org/10.1038/345686a0
  23. Scholer HR, Ruppert S, Suzuki N, Chowdhury K and Gruss P. 1990. New type of POU domain in germ line-specific protein Oct-4. Nature 344:435-439 https://doi.org/10.1038/344435a0
  24. Shiota K, Kogo Y, Ohgane J, Imamura T, Urano A, Nishino K, Tanaka S and Hattori N. 2002. Epigenetic marks by DNA methylation specific to stem, germ and somatic cells in mice. Genes. Cells 7:961-969 https://doi.org/10.1046/j.1365-2443.2002.00574.x
  25. Spencer DS, Ross JW, Ashworth MD, Geisert RD and Rickords LF. 2006. Porcine conceptus Oct-4 mRNA expression during peri-implantation development. Reprod. Domest. Anim. 41:571-572 https://doi.org/10.1111/j.1439-0531.2006.00694.x
  26. Thomas L, Wallace JM, Aitken RP, Mercer JG, Trayhurn P and Hoggard N. 2001. Circulating leptin during ovine pregnancy in relation to maternal nutrition, body composition and pregnancy outcome. J. Endocrinol. 169:465-476 https://doi.org/10.1677/joe.0.1690465
  27. Tian XC, Xu J and Yang X. 2000. Normal telomere lengths found in cloned cattle. Nat. Genet. 26:272-273 https://doi.org/10.1038/81559
  28. van Eijk MJ, van Rooijen MA, Modina S, Scesi L, Folkers G, van Tol HT, Bevers MM, Fisher SR, Lewin HA, Rakacolli D, Galli C, de Vaureix C, Trounson AO, Mummery CL and Gandolfi F. 1999. Molecular cloning, genetic mapping, and developmental expression of bovine POU5F1. Biol. Reprod. 60:1093-1103 https://doi.org/10.1095/biolreprod60.5.1093
  29. Vigneault C, McGraw S, Massicotte L and Sirard MA. 2004. Transcription factor expression patterns in bovine in vitroderived embryos prior to maternal-zygotic transition. Biol. Reprod. 70:1701-1709 https://doi.org/10.1095/biolreprod.103.022970
  30. Wakayama T, Perry AC, Zuccotti M and Johnson KR, Yanagimachi R. 1998. Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei. Nature 394:369-374 https://doi.org/10.1038/28615
  31. Wells DN, Misica PM and Tervit HR. 1999. Production of cloned calves following nuclear transfer with cultured adult mural granulosa cells. Biol. Reprod. 60:996-1005 https://doi.org/10.1095/biolreprod60.4.996
  32. Wilmut I, Schnieke AE, McWhir J, Kind AJ and Campbell KH. 1997. Viable offspring derived from fetal and adult mammalian cells. Nature 385:810-813 https://doi.org/10.1038/385810a0
  33. Yeom YI, Fuhrmann G, Ovitt CE, Brehm A, Ohbo K, Gross M, Hubner K and Scholer HR. 1996. Germline regulatory element of Oct-4 specific for the totipotent cycle of embryonal cells. Development 122:881-894
  34. 고응규, 김종무, 김동훈, 차병현, 황성수, 양병철, 임기순, 김명직, 민관식, 성환후, 2008. 소 착상 전 초기수정란에서 Oct-4 유전자 Promoter 영역의 DNA 메탈화 변화. 한국가축번식학회지 32(1):33-38