Reduced oxygen concentration during in vitro oocyte maturation alters global DNA methylation in the maternal pronucleus of subsequent zygotes in cattle.

Abstract

Preimplantation epigenetic reprogramming is sensitive to the environment of the gametes and the embryo. In vitro maturation (IVM) of bovine oocytes is a critical step of embryo in vitro production procedures and several factors influence its efficiency, including atmospheric oxygen tension. The possibility that the IVM environment can alter this process is tested by determining whether the global DNA methylation pattern (measured via immunofluorescent labeling of 5-methylcytosine [5meC]) in the parental pronuclei of bovine zygotes produced from cumulus-oocyte complexesmatured under low (5%) and atmospheric (~20%) oxygen tension. Normalized 5meC signals differed significantly between maternal and paternal pronuclei of oocytes matured in vitro at 5% oxygen (p ≤ 0.05). There was a significant difference of 5meC between maternal pronuclei of oocytes matured at 5% oxygen and 20% oxygen ( p ≤ 0.05). The relative methylation level (normalized fluorescence intensity of paternal pronucleus divided by the normalized fluorescence intensity of maternal pronucleus) subsequent to maturation in vitro at 5% and 20% oxygen was also significantly altered ( p ≤ 0.05). Our results show that the pattern of global DNA methylation in the maternal pronucleus of bovine zygotes is affected by maturing the oocytes under low oxygen tension which may have an impact on early embryonic development. These data may contribute to the understanding of possible effects of IVM conditions on pronucleus reprogramming.