The effect of Tempo and MitoTEMPO on oocyte maturation and subsequent embryo development in bovine model

Reactive oxygen species (ROS) are one of the factors which reduces oocyte quality and viability of the in vitro produced embryos. Oocyte mitochondria are the major source of ROS production, hence, and the addition of mitochondrion-specific antioxidants could be suggested to minimize the damage caused by ROS during culture. MitoTEMPO, a targeted mitochondrial antioxidant, is formed by conjugating TEMPO to triphenylphosphonium and has an activity like that of superoxide dismutase. It can pass through lipid bilayers easily and accumulate selectively in mitochondria. The goal of this study was to investigate the effects of MitoTEMPO and its non-targeted form, TEMPO, on the developmental competence of bovine oocytes. Accordingly, oocytes were cultured in maturation medium supplemented with either five mM TEMPO (T5) or one μM MitoTEMPO (M1), or T5 + M1 (MT15), or without the antioxidants (C). Nuclear maturation to metaphase II (MII) stage, intracellular glutathione (GSH) content and ROS levels in matured oocytes were analyzed. In addition, cleavage after in vitro fertilization, and blastocyst rates, total cell number in blastocysts as well as the relative abundance of apoptosis-related genes (BAX and BCL2) in blastocysts were determined. Results revealed that the proportion of oocytes at the MII stage, embryos at the blastocyst stage and total cell number in blastocysts increased significantly in the M1 group compared to the C and T5 groups. The levels of intracellular GSH and ROS in oocytes decreased in the M1 group than in the C group (P < 0.05). The expression level of the pro-apoptotic gene (BAX) reduced in blastocysts from the M1 group in comparison to the C and T5 groups (P < 0.05). On the other hand, the expression level of anti-apoptotic gene (BCL2) in obtained blastocysts was not affected by TEMPO and MitoTEMPO. However, the ratio of BAX/BCL2 in blastocysts from the M1 and MT15 groups decreased significantly compared to the C group. These findings suggest that MitoTEMPO can mitigate the adverse effects of oxidative stress on the developmental competence of bovine oocytes.