S-nitroso-N-acetyl penicillamine inhibits spontaneous exit from metaphase-II arrest in rat eggs cultured in vitro.

Abstract

Present study was designed to investigate the in vitro effects of nitric oxide (NO) donor such as S-nitroso-N-acetyl penicillamine (SNAP) on spontaneous exit from metaphase-II arrest (SEM-II) in rat eggs cultured in vitro. Ovulated eggs were denuded and then exposed to various concentrations (0.0, 0.01, 0.1 and 1.0mM) of SNAP for 3h under in vitro culture conditions. The percentage of SEM-II, specific and total phosphorylated cyclin-dependent kinase-1 (Cdk1), cyclin B1 and anaphase promoting complex/cyclosome (APC/C) levels as well as Cdk1 activity were analyzed. The SEM-II was associated with a decrease of Thr-161 phosphorylated Cdk1 as well as cyclin B1 levels and increase of Thr-14/Tyr-15 phosphorylated Cdk1, APC/C levels and Cdk1 activity in aged eggs cultured in vitro. On the other hand, SNAP treatment prevented a decrease of Thr-161 phosphorylated Cdk1 as well as cyclin B1 levels and increase of Thr-14/Tyr-15 phosphorylated Cdk1, Cdk1 activity that finally prevented SEM-II in a concentration-dependent manner. However, APC/C level was not affected by SNAP during the course of treatment in vitro. Present data suggest that SNAP prevented SEM-II possibly by increasing high level of NO and thereby maturation promoting factor (MPF) stabilization in rat eggs cultured in vitro. Hence, SNAP could be used to prevent SEM-II that reduces reproductive outcome in several mammalian species.