Pre-IVM treatment with C-type natriuretic peptide in the presence of cysteamine enhances bovine oocytes antioxidant defense ability and developmental competence in vitro

Abstract

Background: The use of C-type natriuretic peptide (CNP) combined with cysteamine during pre-in vitro maturation (IVM) can help establish an effective pre-IVM system. Aims: This study was designed to investigate the effects of CNP combined with cysteamine on meiotic maturation and developmental competence of bovine oocytes in vitro. Methods: Oocytes were cultured in pre-IVM medium supplemented with 100 nM CNP for 6 h in the absence or presence of 100 μM cysteamine, followed by standard IVM for 24 h. Oocytes matured by standard IVM for 24 h (no pre-IVM) were used as the control. The percentage of oocytes arrested at germinal vesicle (GV) stage in each treatment group was examined at 0 h of IVM, and the nuclear maturation rate was assessed by evaluating the percentages of oocytes that reached metaphase II (M II) stage at 24 h of IVM. After 24 h of IVM, oocytes were subjected to in vitro fertilization (IVF) and in vitro embryo culture (IVC). Cleavage rates were assessed 48 h post-insemination. Blastocyst production rates were recorded on day 8 after IVF. In addition, intra-oocyte glutathione (GSH) and reactive oxygen species (ROS) content for each treatment were tested at 0 h and 24 h of IVM. Results: No differences were observed between the CNP pre-IVM treatment and control groups in the rate of bovine oocytes maturing to the M II stage, cleavage and blastocysts production rates after IVF, and intra-oocyte GSH levels. Notably, the presence of cysteamine during pre-IVM culture with CNP significantly improved the rate of embryos developed to the blastocyst stage after in vitro maturation and fertilization, moreover, it increased the levels of GSH and reduced the levels of ROS in bovine oocytes. Conclusion: The improvement to IVM bovine oocyte developmental competence through pre-IVM with CNP combined with cysteamine may be associated with an increased antioxidant defense. Therefore, such an approach may be a good option for establishing a pre-IVM system.