Autophagy inhibition of immature oocytes during vitrification-warming and in vitro mature activates apoptosis via caspase-9 and-12 pathway

Objective: The aim of this study is to determine the role of autophagy in the immature oocytes during vitrification-warming and in vitro maturation (IVM); the correlations among autophagy, apoptosis, and the activities of caspase in the immature oocytes during vitrification-warming and IVM were also explored. Study design: Immature oocytes from mice were vitrified-warmed and IVM. An autophagy inhibitor (3-methyladenine) was supplemented in cryopreservation solutions and warming solutions. The expression of beclin-1 (an autophagy marker), caspase-3, -8, -9, and -12 were measured. Moreover, the viability of vitrified-warmed immature oocytes and their subsequent developmental competence were measured. Results: The levels of beclin-1 expression in both mRNA and protein in oocytes experienced vitrification warming and IVM were significantly higher than that in fresh immature oocytes experienced IVM. The levels of caspase-3, -9, -12 expression in both mRNA and protein in oocytes vitrified with 3-methyladenine were significantly higher than that vitrified without 3-methyladenine. However, the differences in the caspase-8 expression in both mRNA and protein between the oocytes vitrified with 3-methyladenine and that vitrified without 3-methyladenine were not significant. Conclusion: Immature oocyte cryopreservation exhibits autophagic activation. Autophagy inhibition of the immature oocytes during vitrification-warming and IVM activates apoptosis via caspase-9 and -12 pathway. (C) 2017 Published by Elsevier Ireland Ltd.