The effects of metabolic substrates glucose, pyruvate, and lactate added to a skim milk-based semen extender for cooled storage of stallion sperm.
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Under invitro conditions, stallion sperm might preferentially use energy substrates that primarily undergo mitochondrial metabolism. The present study sought to determine the effects of glucose, pyruvate, lactate, or their combinations on the quality of stallion sperm subjected to cooled storage at different temperatures, when using a skim milk-based semen extender. In Experiment 1, no substrate (Control), glucose (40mM; Glu-40), pyruvate (2mM, 19.8mM; Pyr-2, Pyr-19), lactate (2mM, 19.8mM; Lac-2, Lac-19, respectively), or their combinations (G/P/L-2 or G/P/L-19, respectively) were added to a milk-based extender and their effects were determined on motion characteristics, viability/acrosomal intactness (VAI), lipid peroxidation status (VLPP), and DNA integrity (COMP-t) of sperm incubated for 1h at 37C, or sperm stored for 24 h at either 10 or 20C. At any period and temperature tested, Glu-40, G/P/L-2, and G/P-L-19 resulted in similar motion characteristics (P>0.05) but were higher than that of other treatment groups (P<0.05). Mean VAI was highest in Glu-40 (P<0.05). Mean VLPP was highest in G/P/L-2 and G/P/L-19 groups (P<0.05), and mean COMP-t was lowest in Control, Glu-40, G/P/L-2 and G/P/L-19 groups (P<0.05). All measures of sperm quality were higher in semen stored at 10C than 20C (P<0.05). In Experiment 2, increasing concentrations of either pyruvate or lactate (Pyr-40, Lac-40 or Pyr-80, Lac-80) were added to the extender as energy substrates and compared to glucose (40mM), following storage for 72h at either 10 or 20C. Groups Glu-40 and Pyr-40 yielded similar sperm motion characteristics and VAI, while VLPP and COMP-t were reduced in these treatment groups, as compared to Pyr-80, Lac-40, and Lac-80 (P<0.05). All measures of sperm quality were higher in semen stored at 10C vs 20C (P<0.05). This study demonstrates that at storage temperatures of 10 or 20C, stallion sperm quality is optimized by the presence of glucose in a skim milk-based semen extender. The addition of substrates that readily support oxidative phosphorylation (i.e., pyruvate or lactate) did not improve the quality of stallion sperm over that of glucose alone and resulted in deleterious effects on sperm quality over time. These effects appeared to be associated with oxidative stress. Use of pyruvate (40mM) as an alternative energy substrate to glucose generally yielded similar results to that of glucose when sperm were stored at 10C only.
Hernndez-Avils, C., Ramrez-Agmez, L., Love, C. C., Friedrich, M., Pearson, M., Kelley, D. E., ... Varner, D. D.
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Hernández-Avilés, Camilo||Ramírez-Agámez, Luisa||Love, Charles C||Friedrich, Macy||Pearson, Mariah||Kelley, Dale E||Beckham, Anne MN||Teague, Sheila R||LaCaze, Katrina A||Brinsko, Steven P||Varner, Dickson D
