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Previous studies have shown that melatonin can mitigate cryopreservation-induced mitochondrial dysfunction in oocytes; however, the underlying molecular mechanism remains unclear. The objective of the present study was to investigate whether melatonin can improve the mitochondrial function during  in vitro  maturation of vitrified-warmed mouse germinal vesicle (GV) oocytes by modulating phosphorylation of dynamin related protein 1 (Drp1). Vitrification/warming procedures resulted in the following: (1) After cryopreservation of mouse GV oocytes, the phosphorylation level of Drp1 at Ser616 (p-Drp1 Ser616) in metaphase II (MII) oocytes was increased ( P  <  0.05 ). Furthermore, the rates of  in vitro  maturation, cleavage and blastocyst formation after parthenogenetic activation were decreased ( P  <  0.05 ). (2) In MII oocytes, the expression levels of translocase of the mitochondrial outer membrane 20 (TOMM20), mitochondrial membrane potential (MMP), adenosine triphosphate (ATP) content, and mRNA levels of mitochondrial biogenesis-related genes ( Sirt1, Pgc-1 α , Tfam ) were all decreased ( P  <  0.05 ), and (3) Reactive oxygen species (ROS) level, early apoptosis level, Cytochrome C release and mRNA levels of pro-apoptotic related genes ( Bax, Caspase9, Caspase3 ) in MII oocytes were all increased ( P  <  0.05 ). The results of this study further revealed that negative impacts of GV oocyte cryopreservation were mitigated by supplementation of warming and  in vitro  maturation media with 10 −7 mol /L melatonin or 2 x 10 −5 mol/L Mdivi-1 (Drp1 inhibitor). Therefore, we concluded that 10 −7 mol/L melatonin improved mitochondrial function, reduced oxidative stress and inhibited apoptosis by regulating phosphorylation of Drp1, thereby enhancing  in vitro  development of vitrified-warmed mouse GV oocytes.

Melatonin Promotes in vitro Development of Vitrified-Warmed Mouse GV Oocytes, Potentially by Modulating Phosphorylation of Drp1