Active loss of DNA methylation in two-cell stage goat embryos

Abstract

Early mammalian embryos are thought to gain nuclear totipotency through DNA methylation reprogramming (DMR). By this process, DNA methylation patterns acquired during gametogenesis that are unnecessary for zygotic development are erased. The DMR patterns of various mammalian species have been studied; however, they do not seem to have a conserved pattern. We examined early goat embryos to find conforming rules underlying mammalian DMR patterns. Immunocytochemical results showed that the overall level of DNA methylation was not greatly changed during the pronucleus stage. At the two-cell stage, active demethylation occurred and simultaneously affected both parental DNAs, resulting in a global loss of 5-methylcytosine. The level of DNA methylation was lowest in the four-cell stage, with increased de novo methylation during the eight-cell stage. Histone H3-lysine 9 was gradually trimethylated in the sperm-derived chromatin, continuing from the pronucleus stage through the two-cell stage. This goat DMR pattern is novel and distinct from the DMRs of other mammalian species. The more mammalian species we included for DMR analysis, the more multifarious patterns we obtained, adding an extra diversity each time to the known mammalian DMR patterns. Nevertheless, the evolutionary significance and developmental consequence of such diverse DMR patterns are currently unknown.