Paradoxical effects of inhibition of Δ14-reductase and Δ7-reductase on porcine oocyte maturation and subsequent embryo development after parthenogenetic activation

Follicular fluid-derived meiosis-activating sterol (FF-MAS), an intermediate in the cholesterol biosynthesis pathway, plays a crucial role in the meiotic resumption of mammalian oocytes. Maintaining a high concentration of FF-MAS in vitro is challenging; therefore, AY9944 A-7, an inhibitor of z14-reductase [which converts FF-MAS to testis meiosis-activating sterol (T-MAS)] and z7-reductase (which converts T-MAS to cholesterol), has been used to enhance oocyte maturation. This study examined the effects of various concentrations (0, 10, 20, and 40 mu M) of AY9944 A-7 on porcine oocyte maturation and subsequent embryo development. Results indicate that treatment with 10 and 20 mu M AY9944 A-7 during in vitro maturation (IVM) enhanced oocyte nuclear maturation, with 10 mu M significantly increasing the transcript expression of oocyte maturation-related genes. However, blastocyst formation rates significantly decreased in oocytes treated with AY9944 A-7 concentrations above 10 mu M. To explore these unexpected findings, the study evaluated the effects of AY9944 A-7 on lipid content in oocytes and the sonic hedgehog (SHH) signaling pathway in subsequent parthenogenetic embryos. A concentration-dependent decrease in oocyte lipid content was observed following AY9944 A-7 treatment. Additionally, transcripts of SHH signaling pathway genes were detected in preimplantation-stage parthenogenetic embryos, with reduced expression in the 10 mu M AY9944 A-7-treated group. Taken together, AY9944 A-7 supplementation during porcine IVM enhanced oocyte maturation by accumulating FF-MAS, but subsequent embryo development was impaired due to cholesterol deficiency, potentially mediated by SHH signaling downregulation.