初期G1期細胞によるウシ核移植胚におけるDNAのメチル化レベルの検討
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概要
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Although cloning by somatic cell nuclear transfer （NT） has achieved in various mammalian species, its efficiency has been extremely low. Recently, enhanced development to full ‒ term was obtained with NT embryos with bovine early G1 cells rather than NT embryos with serum starved （G0） cells. In this study, we examined genome ‒ wide DNA methylation of cloned bovine embryos with early G1 cells by immunostaining technique with an antibody to 5‒methyl cytosine. Somatic cell nuclear transfer embryos were produced by bovine enucleated matured oocytes electrofusing with bovine G0‒ or eG1 ‒ phase fibroblasts. Levels of DNA methylation of the NT embryos both with G0 of eG1 cells were compared to those of in ‒ vitro fertilized embryos at 8‒ cell stage. Our results indicated that regardless of the donor cell type, greater relative levels of DNA methylation were observed in NT embryos than those of IVF embryos. However, the relative levels of DNA methylation of eG1‒NT embryos were lower than those of G0‒NT embryos. Thus, eG1‒NT embryos were genome ‒ widely hypomethylated compared with G0‒NT embryos. These results suggest that a possible reason of the high success rate of eG1‒NT embryos may be due to their lower methylation levels than those of G0‒NT embryos at 8‒cell stage.