Pyruvate enhances stallion sperm function in high glucose media improving overall metabolic efficiency

Abstract

If a mechanism of more efficient glycolysis depending on pyruvate is present in stallion spermatozoa, detrimental effects of higher glucose concentrations that are common in current commercial extenders could be counteracted. To test this hypothesis, were incubated in a 67 mM Glucose modified Tyrode’s media in the presence of 1- or 10-mM pyruvate and in the Tyrode’s basal media which contains 5 mM glucose. incubated for 3 h at 37 ◦C in 67 mM Tyrode’s media with 10 mM pyruvate showed increased in comparison with aliquots incubated in Tyrode’s 5 mM glucose and Tyrode’s 67 mM glucose (57.1 ± 3.5 and 58.1 ± 1.9 to 73.0 ± 1.1 %; P < 0.01). Spermatozoa incubated in Tyrode’s with 67 mM glucose 10 mM pyruvate maintained the viability along the 64.03 ± 15.4 vs 61.3 ± 10.2), while spermatozoa incubated in 67 mM Glucose-Tyrode’s a decrease in viability (38.01 ± 11.2, P < 0.01). 40 mM oxamate, an inhibitor of the lactate dehydrogenase LDH, reduced sperm viability (P < 0.05, from 76 ± 5 in 67 mM Glucose/10 mM pyruvate to 68.0 ± 4.3 %, P < 0.05). Apoptotic markers increased in the presence of oxamate. (P < 0.01). UHPLC/MS/MS showed that 10 mM pyruvate pyruvate, lactate, ATP and NAD+ while phosphoenolpyruvate decreased. The that explain the improvement of in presence of 10 mM pyruvate involve the conversion of lactate to pyruvate and increased NAD+ enhancing the efficiency of the glycolysis.