Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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DNA Methylation Change of Oct-4 Gene Promoter Region during Bovine Preimplantation Early Embryos

소 착상 전 초기수정란에서 Oct-4 유전자 Promoter 영역의 DNA 메틸화 변화

  • 고응규 (농촌진흥청 축산과학원 응용생명공학과) ;
  • 김종무 (충북대학교 축산학과) ;
  • 김동훈 (농촌진흥청 축산과학원 응용생명공학과) ;
  • 차병현 (농촌진흥청 축산과학원 응용생명공학과) ;
  • 황성수 (농촌진흥청 축산과학원 응용생명공학과) ;
  • 양병철 (농촌진흥청 축산과학원 응용생명공학과) ;
  • 임기순 (농촌진흥청 축산과학원 응용생명공학과) ;
  • 김명직 (농촌진흥청 축산과학원 응용생명공학과) ;
  • 민관식 (한경대학교 생물환경정보통신전문대학원) ;
  • 성환후 (농촌진흥청 축산과학원 응용생명공학과)
  • Published : 2008.03.31
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Abstract

DNA methylation is involved in tissue-specific gene control and essential for normal embryo development Octamer-binding transcription factor 4 (Oct-4) is one of the most important transcription factors for early differentiation. This study was performed whether the bovine Oct-4 is tissue specific or developmental dependent epigenetic mark, we investigated transcripts and the methylation status of CpGs of 5'-promoter region of Oct-4 in bovine preimplantation embryos. Oct-4 transcripts were highly detected in morula and blastocyst, while they were present low levels in sperm and 2- to 8-cell stage embryos. These results suggest that de novo expression of Oct-4 initiates at morula stage of embryogenesis. Here we determined that there is a tissue-dependent differentially methylated region (T-DMR) in the 5'-promoter region of Oct-4. The methylation status of the Oct-4 T-DMR was distinctively different in the oocyte from that in the sperm and adult somatic tissues and changed from zygote to blastocyst stage, suggesting that active methylation and demethylation occur during preimplantation development. Based on these results, the 5'-promoter region of Oct-4 gene is target for DNA methylation and the methylation status changes variously during embryonic development in bovine.

DNA 메틸화는 조직특이적인 유전자 조절에 관여하고, 정상적인 배 발달에 필수적이다. POU5F1은 octamer-binding transcription factor 4 (Oct-4)를 encode하며, 초기 분화에 중요한 전사인자이다. 본 실험에서 소의 Oct-4가 조직특이적이고 발달의존적인 epigenetic 표지 인지를 검토하고자, 착상 전 수정란에서 Oct-4 전사산물과 상류 promoter 영역의 CpGs의 메틸화를 조사하였다. Oct-4 전사산물은 정자 그리고 2-cell에서 8-cell 수정란까지 낮은 수준으로 존재하지만, 상실배와 배반포에서 높게 검출되었다. 이러한 결과는 배 발달 과정의 상실배 단계에서 Oct-4의 de novo 발현이 시작됨을 의미한다. Oct-4 상류 promoter 영역에는 메틸화 가변 영역 (tissue-dependent differentially methylated region, T-DMR)이 존재한다. Oct-4 메틸화 가변 영역의 메틸화 상태는 정자, 성체 체조직과 난자에서 서로 다르고, 수정란으로부터 배반포 단계까지 변화하였는데, 이는 착상 전 초기 배 발달 과정에 active 메틸화와 탈메틸화가 일어남을 의미한다. 이상의 결과, Oct-4 유전자 상류 promoter 영역은 DNA 메틸화의 타깃이고, 그 메틸화 상태는 소 수정란 발달 동안에 다양하게 변화한다.

Keywords

References

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