Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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Dynamic DNA Methylation Change of Dnmt1o 5'-Terminal Region during Preimplantation Development of Cloned Pig

돼지 체세포 복제란 초기발달 과정 중 Dnmt1o 상류 영역의 다이내믹한 DNA 메틸화 변화

  • 고응규 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 김성우 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 조상래 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 도윤정 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 김재환 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 김상우 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 김현 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 박재홍 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 박수봉 (농촌진흥청 국립축산과학원 가축유전자원시험장)
  • Received : 2012.02.26
  • Accepted : 2012.03.26
  • Published : 2012.03.31
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Abstract

DNA methyltransferase 1 (Dnmt1) gene contains three different isoform transcripts, Dnmt1s, Dnmt1o, and Dnmt1p, are produced by alternative usage of multiple first exons. Dnmt1o is specific to oocytes and preimplantation embryos, whereas Dnmt1s is expressed in somatic cells. Here we determined that porcine Dnmt1o gene had differentially methylated regions (DMRs) in 5'-flanking region, while those were not found in the Dnmt1s promoter region. The methylation patterns of the porcine Dnmt1o/Dnmt1s DMRs were investigated using bisulfite sequencing and pyrosequencing analysis through all preimplantation stages from one cell to blastocyst stage in in vivo or somatic cell nuclear transfer (SCNT). The Dnmt1o DMRs contained 8 CpG sites, which located in -640 bp to -30 bp upstream region from transcription start site of the Dnmt1o gene. The methylation status of 5 CpGs within the Dnmt1o DMRs were distinctively different at each stage from one-cell to blastocyst stage in the $in$ $vivo$ or SCNT, respectively. 55.62% methylation degree of the Dnmt1o DMRs in the $in$ $vivo$ was increased up to 84.38% in the SCNT embryo, moreover, $de$ $novo$ methylation and demethylation occurred during development of porcine embryos from the one-cell stage to the blastocyst stage. However, the DNA methylation states at CpG sites in the Dnmt1s promoter regions were hypomethylated, and dramatically not changed through one-cell to blastocyst stage in the $in$ $vivo$ or SCNT embryos. In the present study, we demonstrated that the DMRs in the promoter region of the porcine Dnmt1o was well conserved, contributing to establishment and maintenance of genome-wide patterns of DNA methylation in early embryonic development.

Keywords

References

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