Problem statement:To identify the metabolic reaction-glycolysis or oxidative phosphorylation that is mainly involved in the production of energy required for rat sperm mobilization. Approach: Epididymal sperm were collected from Wistar rats and extended in lactatecontaining or lactate-free raffinose-modified Krebs-Ringer Bicarbonate solution (mKRB)-egg yolk medium supplemented with 0, 1, 2, or 3 mM 2-Deoxy-D-Glucose (2 DG) and 1, 2, or 3 mM sodium Oxamate (OX). Sperm motility, straight-line velocity (VSL) and oxygen consumption were evaluated. Further, immunofluorescent localization of Lactate Dehydrogenase C (LDH-C) in sperm was also performed. Results: Low concentrations of 2DG (1 and 2 mM) did not significantly affect motility, VSL and oxygen consumption of sperm extended in the lactate-containing medium. While sperm motility and oxygen consumption were significantly inhibited by even 1mM 2DG in sperm extended in lactate-free medium. Sperm motility significantly inhibited in the case of sperm extended in lactatecontaining and free-medium with 1 mM OX. We also found that sperm motility was not maintained in the absence of lactate throughout the 3 h incubation period. Immunofluorescence study revealed that mainly LDH-C was may be localized in the intramitochondrial region of the sperm. Conclusion: These results suggest that exogenous lactate enhances lactate oxidation by LDH-C, thereby promoting mitochondrial oxidative reactions in the midpiece and maintaining the mobilization of rat epididymal sperm.
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