Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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DNA Methylation Change of Repeats Sequences in Pig SCNT Embryos Produced under Different Osmolarity Culture Conditions

삼투압 배양 조건에 따른 돼지 체세포 복제 배반포에서 Repeats 영역의 DNA 메틸화 변화

  • Ko, Yeoung-Gyu (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Im, Gi-Sun (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Park, Mi-Rung (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Woo, Jae-Seok (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Yang, Byoung-Chul (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Hwang, Seong-Soo (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Lee, Hwi-Cheul (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Lee, Poong-Yeon (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Cho, Chang-Yeon (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Choi, Sun-Ho (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Yoo, Young-Hee (Animal Genetic Resources Station, National Institute of Animal Science, RDA)
  • 고응규 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 임기순 (농촌진흥청 국립축산과학원 동물바이오공학과) ;
  • 박미령 (농촌진흥청 국립축산과학원 동물바이오공학과) ;
  • 우제석 (농촌진흥청 국립축산과학원 동물바이오공학과) ;
  • 양병철 (농촌진흥청 국립축산과학원 동물바이오공학과) ;
  • 황성수 (농촌진흥청 국립축산과학원 동물바이오공학과) ;
  • 이휘철 (농촌진흥청 국립축산과학원 동물바이오공학과) ;
  • 이풍연 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 조창연 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 최순호 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 유용희 (농촌진흥청 국립축산과학원 가축유전자원시험장)
  • Received : 2010.08.30
  • Accepted : 2010.09.15
  • Published : 2010.09.30
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Abstract

Osmolarity of culture media is one of the most important factors affecting in vitro development. This study was conducted to investigate the DNA methylation status of Pre-1 and satellite sequence in pig nuclear transfer (pNT) embryos produced under different osmolarity culture conditions. Control group of pNT embryos was cultured in PZM-3 for six days. Other two treatment groups of pNT embryos were cultured in modified PZM-3 with 138 mM NaG or 0.05M sucrose (mPZM-3, 320 mOsmol) for two days, and then cultured in PZM-3 (270 mOsmol) for four days. Previous our studies have reported that pNT embryos cultured in both hypertonic media showed significantly higher blastocyst formation rate than that of control. The DNA methylation status of the satellite sequences in blastocyst was characterized using bisulfite-sequencing technology. The satellite region had a similar methylation pattern of in vivo blastocyst among two culture groups excepting the control group. Each level of methylation is that the satellite DNA moderately methylated (43.10% of PZM-3; 56.12% of NaCl; 55.06% of sucrose; 60.00% of in vivo embryos). As a result of the sequence of PRE-1, CpG methylation pattern was similar to three groups, including in vivo group. In case of the satellite DNA region, the osmolarity conditions were affected CpG DNA methylation status while PRE-1 sequence was not affected CpG DNA methylation in pNT blastocyst stage. These results indicate that the modification of osmolarity in a culture media may influence to spatially change of DNA methylation of repetitive sequence for pNT embryo development.

Keywords

References

  1. 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
  2. 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.
  3. Hwang IS, Park MR, Moon HJ, Shim JH, Kim DH, Yang Be, Ko YG, Yang BS, Cheong HT, Im GS (2007): Osmolarity at early culture stage affects development and expression of apoptosis related genes (Bax-$\alpha$ and Bcl-xl) in preimplantation porcine NT embryos. Mol Reprod Dev 75:464-471.
  4. Im GS, Yang BS, Lai L, Liu Z, Hao Y, Prather RS (2005): Fragmentation and development of preimplantation porcine embryos derived by parthenogenetic activation and nuclear transfer. Mol Reprod Dev 71:159-165. https://doi.org/10.1002/mrd.20258
  5. Kafri T, Gao X, Razin A (1993): Mechanistic aspects of genome-wide demethylation in the preimplantation mouse embryo. Proc Natl Acad Sci USA 90:10558-10562. https://doi.org/10.1073/pnas.90.22.10558
  6. Kang YK, Koo DB, Park JS, Choi YH, Kim HN, Chang WK, Lee KK, Han YM (2001): Typical demethylation events in cloned pig embryos. J Biol Chem 276:39980-39984. https://doi.org/10.1074/jbc.M106516200
  7. Kim SH, Kang YK, Koo DB, Kang MJ, Moon SJ, Lee KK, Han YM (2004): Differential DNA methylation reprogramming of various repetitive sequences in mouse preimplantation embryos. Biochemical and Biophysical Research Communications 324:58-63. https://doi.org/10.1016/j.bbrc.2004.09.023
  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. Kremenskoy M, Kremenska Y, Suzuki M, Imai K, Takahashi S, Hashizume K, Yagi S, 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
  10. Ohgane J, Wakayama T, Kogo Y, Senda S, Hattori N, Tanaka S, Yanagimachi R, Shiota K (2001): DNA methylation variation in cloned mice. Genesis 30:45-50. https://doi.org/10.1002/gene.1031
  11. Park MR, Hwang IS, Shim JH, Moon HJ, Kim DH, Ko YK, Seong HH, Im GS (2008): Effect of osmolarity of culture medium on imprinting and apoptotic gene expression in miniature pig nuclear transfer embryos. Reprod Dev Biol 32(3):183-191.
  12. Reik W, Dean W, Walter J (2001): Epigenetic reprogramming in mammalian development. Science 293:1089-1093. https://doi.org/10.1126/science.1063443
  13. Santos F, Hendrich B, Reik W, Dean W (2002): Dynamic reprogramming of DNA methylation in the early mouse embryo. Dev Biol 241:172-Z182. 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
  15. Shi W, Haaf T (2002): Aberrant methylation patterns at the two-cell stage as an indicator of early developmental failure. Mol Reprod Dev 63:329-334. https://doi.org/10.1002/mrd.90016
  16. Yagi S, Hirabayashi K, Sato S, Li W, Takahashi Y, Hirakawa T, Wu G, Hattori N, Hattori N, Ohgane J (2008) DNA methylation profile of tissue-dependent and differentially methylated regions (T-DMRs) in mouse promoter regions demonstrating tissue-specific gene expression. Genome Res 18:1969-1978. https://doi.org/10.1101/gr.074070.107