Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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DNA Methylation Change of Dnmt1o and Dnmt1s 5'-Region in the Early Porcine Embryo

돼지 초기수정란에서 Dnmt1o와 Dnmt1s 상류 영역의 DNA 메틸화 변화

  • Kim, Hyun-Mi (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Kim, Sung-Woo (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Cho, Sang-Rae (Subtropical Genetics Resources Station, National Institute of Animal Science, RDA) ;
  • Kim, Hyun (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Park, Jae-Hong (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Cho, Jae-Hyeon (Institute of Life Science, College of Veterinary Medicine, Gyeongsang National University) ;
  • Yang, Boh-Suk (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Ko, Yeoung-Gyu (Animal Genetic Resources Station, National Institute of Animal Science, RDA)
  • 김현미 (농촌진흥청 국립축산과학원 동물바이오공학과) ;
  • 김성우 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 조성래 (농촌진흥청 국립축산과학원 난지축산시험장) ;
  • 김현 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 박재홍 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 조재현 (경상대학교 수의과대학) ;
  • 양보석 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 고응규 (농촌진흥청 국립축산과학원 가축유전자원시험장)
  • Received : 2011.08.18
  • Accepted : 2011.08.25
  • Published : 2011.09.30
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Abstract

In the present study, we identified differentially methylated region (DMR) upstream of Dnmt1o and Dnmt1s gene in early porcine embryos. Porcine Dnmt1o had at least one DMR which was located between -530 bp to -30 bp upstream from transcription start site of the Dnmt1o gene. DNA methylation analyses of Dnmt1o revealed the DMR to be hypomethylated in oocytes, whereas it was highly methylated in sperm. Moreover, the DMR upstream of Dnmt1o was gradually hypermethylated from oocytes to two cells and dramatically changed in the methylation pattern from four cells to BL stages in an in vivo. In an IVF, the methylation status in the DMR upstream of Dnmt1o was hypermethylated from one cell to eight cells, but demethylated at the Morula and BL stages, indicating that the DNA methylation pattern in the Dnmt1o upstream ultimately changed from stage to stage before the implantation. Next, to elucidate whether DNA methylation status of Dnmt1s upstream is stage-by-stage changed in during porcine early development, we analyzed the dynamics of the DNA methylation status of the Dnmt1s locus in germ cell, or one cell to BL cells. The Dnmt1s upstream was highly methylated in one and eight cells, while less methylated in two, four, morula, and BL cells. Taken together, our data demonstrated that DNA methylation and demethylation events in upstream of Dnmt1o/Dnmt1s during early porcine embryos dramatically occurred, and this change may contribute to the maintenance of genomewide DNA methylation in early embryonic development.

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, Meehan, RR (2004): The effect of interspecific oocytes on demethylation of sperm DNA. Proc Natl Acad Sci USA 101:7636-7640. https://doi.org/10.1073/pnas.0400730101
  2. Bestor TH (2000): The DNA methyltransferases of mammals. Hum Mol Genet 9:2395-2402. https://doi.org/10.1093/hmg/9.16.2395
  3. Chen T, Li E (2004): Structure and function of eukaryotic DNA methyltransferases. Curr Top Dev Biol 60:55-89.
  4. Clark SJ, Harrison J, Paul CL, Frommer M (1994): High sensitivity mapping of methylated cytosines. Nucleic Acids Res 22:2990-2997. https://doi.org/10.1093/nar/22.15.2990
  5. Dean W, Santos F, Stojkovic M, Zakhartchenko V, Walter J, Wolf E, Reik W (2001): Conservation of methylation reprogramming in mammalian development: aberrant reprogramming in cloned embryos. Proc Natl Acad Sci USA 98:13734-13738. https://doi.org/10.1073/pnas.241522698
  6. Fulka H, Mrazek M, Tepla O, Fulka JJr (2004): DNA methylation pattern in human zygotes and developing embryos. Reproduction 128:703-708. https://doi.org/10.1530/rep.1.00217
  7. Jeong YS, Dh KB, Park JS, Kim JS, Kang YK (2009): Cytoplasmic localization of oocyte-specific variant of porcine DNA methyltransferase-1 during early development. Dev Dyna 238:1666-1673. https://doi.org/10.1002/dvdy.21975
  8. Ko YG, Nishino K, Hattori N, Arai Y, Tanaka S, Shiota K (2005): Stage-by-stage change in DNA methylation status of Dnmt1 locus during mouse early development. J Biol Chem 280:9627-9634.
  9. Li E, Bestor TH, Jaenisch R (1992): Targeted mutation of the DNA methyltransferase gene results in embryonic lethality. Cell 69:915-26. https://doi.org/10.1016/0092-8674(92)90611-F
  10. Mertineit C, Yoder JA, Taketo T, Laird DW, Trasler JM, Bestor TH (1998): Sex-specific exons control DNA methyltransferase in mammalian germ cells. Development 125:889-897.
  11. Reik W, Dean W, Walter J (2001): Epigenetic reprogramming in mammalian development. Science 293: 1089-1093. https://doi.org/10.1126/science.1063443
  12. Rougier N, Bourc'his D, Gomes DM, Niveleau A, Plachot M, Paldi A, Viegas-Pequignot E (1998): Chromosome methylation patterns during mammalian preimplantation development. Genes Dev 12:2108- 2113. https://doi.org/10.1101/gad.12.14.2108
  13. Santos F, Hendrich B, Reik W, Dean W (2002): Dynamic reprogramming of DNA methylation in the early mouse embryo. Dev Biol 241:172-82. https://doi.org/10.1006/dbio.2001.0501
  14. Shiota K, Kogo Y, Ohgane J, Imamura T, Urano A, Nishino K, Tanaka S, 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