Effect of 2, 4-Dinitrophenol on reactive oxygen species production in mitochondria and improvement of sperm viability after freeze-thawing process of Rooster semen

Regarding the role of mitochondria in the quality of sperm motility and fertility, protection of mitochondrial function against ROS-induced damage can be an important step in improving roster sperm fertility. The mitochondria are the most important source of ROS and the unique properties of mitochondria prevent antioxidants from entering, which results in poor antioxidant performance in protecting ROS-induced damage. Therefore, the use of antioxidants that can pass through the mitochondrial membrane and inhibit ROS in this region can be a good way to reduce the damage caused by ROS and improve the sperm quality and fertility. The aim of the present study was to use mitochondrial mild uncoupling combinations to reduce the risk of ROS-induced injuries and increase the survival of sperm after freeze-thawing. For this purpose, 14 Ross breed rooster with 28 weeks of age were used. The semen was taken by massaging the abdomen-back procedure. After initial evaluation, semen samples were pooled After dilution of semen samples and addition of 2-4-dinitrophenol (DNP) at five levels (0, 0.25, 0.5, 0.75, 1 nM), they were cooled and then frozen. After two months, semen samples were thawed and evaluated for the percentage of dead and live sperm, sperm motility parameters, membrane integrity, lipid peroxidation and sperm morphology. The results of this study showed that addition of DNP at 0.75 nM resulted in improvement in total and progressive motility, viability, plasma membrane integrity and sperm normality compared to the control group, and significantly decreased MDA concentration compared to control group (p <0.05).