Abstract

The efficiency of in vitro embryo production remains low compared with that observed in vivo. Recent studies have independently shown that cyclic adenosine monophosphate (cAMP) modulation prior to in vitro maturation (IVM) supplementation improves oocyte developmental competence. In this context, special cAMP modulators have been applied during IVM as promising alternatives to improve this biotechnology. Accordingly, this study was conducted to evaluate the effects of treatment with cilostazol, a PDE3 inhibitor, during pre-IVM culture on oocyte meiotic maturation in yak. Immature yak cumulus-oocyte complexes (COCs) were treated in vitro without (control) or with 5μM cilostazol for 0, 2, or 4h prior to IVM. Results showed that the presence of cilostazol in pre-IVM medium significantly increased the percentages of oocytes at metaphase II stage compared with that in the control groups (P<0.05). Moreover, pre-IVM with cilostazol significantly enhanced intraoocyte cAMP and glutathione (GSH) levels at the pre-IVM or IVM phase relative to the no pre-IVM groups (P<0.05). After in vitro fertilization (IVF) and parthenogenetic activation (PA), the developmental competences of oocytes and embryo quality were improved significantly after pre-IVM with cilostazol compared with the control groups (P<0.05), given that the cleavage and blastocyst formation rates and the total number of blastocyst cells were increased. The presence of cilostazol also increased the levels of mRNA expression for adenylate cyclase 3 (ADCY3) and protein kinase 1 (PKA1), as well as decreased the abundance of phosphodiesterase 3A (PDE3A) in COCs and IVF blastocysts, compared with their control counterparts (P<0.05). The results demonstrated that the meiotic progression of immature yak oocytes could be reversibly affected by cAMP modulators. By contrast, treatment with cilostazol during pre-IVM positively affected the developmental competence of yak oocytes, probably by improving intraoocyte cAMP and GSH levels and regulating mRNA expression patterns. We concluded that appropriate treatment with cilostazol during pre-IVM would be beneficial for oocyte maturation in vitro.

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