The occurrence of infertility in diabetic patients is attributed to oxidative damage of peroxidized products. High glucose-induced mitochondrial oxidative stress and glycolytic enzyme inactivation is considered to be an important mediator for sperm dysfunction. In this study, we successfully constructed TM3-GAPDS stable strain and investigated the role of sperm specific glyceraldehyde-3-phosphate dehydrogenase (GAPDS) on high glucose-induced apoptosis in TM3 cells. High glucose decreased the protein expression of SOD2 and catalase, while the level of intracellular ROS and the apoptosis - related protein increased in TM3 cells. Furthermore, high glucose-induced oxidative stress and apoptosis were reversed by GAPDS overexpression or antioxidant treatment. In conclusion, our data suggest that GAPDS overexpression antagonize high glucose-induced apoptosis by controlling ROS accumulation in TM3 cells. (C) 2018 Elsevier Inc. All rights reserved.