earticle

영어

Inhibition of MEK1/2 and GSK-3(2i system) is useful for maintenance of navie state of embryonic stem cells in various species. However, the role of 2i system in porcine preimplantation stage embryos is not clear. In this study, we investigated the quality of blastocyst and pluripotent-related factors by 2i system and epigenetic modification status by 2i system during porcine early embryo development. Treatment of MEK1/2 inhibitor PD0325901 (4 uM), GSK-3 inhibitor CHIR99021 (0.3 uM) and 2i (PD0325901+ CHIR99021) did not enhanced the rate of blastocyst formation. However, the size of blastocyst was increased in 2i treated embryos. TUNEL assay showed that CHIR- 99021 and 2i treatment enhanced blastocyst quality. Furthermore, 2i treated blastocyst increased OCT4 and SOX2 positive cells and decreased GATA4 positive cells. Methylation of histone H3 lysin trime3 (h3k9me3) were reduced, and histone H3 deacetylaton (h3k9ac) were increased in 2i treated embryos. Furthermore, bi-sulfite sequencing results showed that 2i treatment groups demethylated in satellite I region. And, RNA methylatransferase protein METTL3 was increased while demethylase FTO was increased after treatment of 2i. In conclusion, our findings strongly suggest that 2i system can be useful method for increasing of blastocyst quality by affecting of ICM/TE formation by regulating epigenetic modification during early embryo development.