The effects of metabolic substrates glucose, pyruvate, and lactate added to a skim milk-based semen extender for cooled storage of stallion sperm.
Abstract: Under in vitro 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 (40 mM; Glu-40), pyruvate (2 mM, 19.8 mM; Pyr-2, Pyr-19), lactate (2 mM, 19.8 mM; 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) of sperm incubated for 1 h at 37 °C, or sperm stored for 24 h at either 10 or 20 °C. 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 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 10 °C than 20 °C (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 (40 mM), following storage for 72 h at either 10 or 20 °C. Groups Glu-40 and Pyr-40 yielded similar sperm motion characteristics and VAI, while VLPP and COMP 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 10 °C vs 20 °C (P < 0.05). This study demonstrates that at storage temperatures of 10 or 20 °C, 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 (40 mM) as an alternative energy substrate to glucose generally yielded similar results to that of glucose when sperm were stored at 10 °C only.
Copyright © 2020 Elsevier Inc. All rights reserved.
Publication Date: 2020-11-30 PubMed ID: 33302166DOI: 10.1016/j.theriogenology.2020.11.017Google Scholar: Lookup
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Summary
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This research explores the impact of various energy substrates like glucose, pyruvate, and lactate, incorporated in a skim milk-based semen extender, on the quality of stored stallion sperm. The study deduces that amid the substrates used, glucose promotes optimal stallion sperm quality during storage at temperatures of 10 or 20 degrees Celsius.
Metabolic Substrates and Stallion Sperm Quality
- The researchers examined the utility of metabolic substrates, namely, glucose, pyruvate, lactate, and their combinations, to enhance the quality of stallion sperm during cooled storage. These substrates were added to a skim milk-based semen extender.
- A pair of experiments were conducted. The first experiment featured the addition of no substrate as a control, glucose, pyruvate, lactate, or their combinations to the extender, and their effects were gauged on sperm quality factors like motion characteristics, viability/acrosomal intactness (VAI), lipid peroxidation status (VLPP), and DNA integrity (COMP).
- Sperm samples that were incubated for an hour at 37⁰C or stored for 24 hours at either 10⁰C or 20⁰C underwent testing. The results showed that glucose and certain combinations of substrates showed better motion characteristics and VAI compared to others.
- This experiment also indicated that sperm quality was better preserved at a lower storage temperature of 10⁰C as compared to 20⁰C.
The Effects of Increasing Concentration of Substrates
- In the second experiment, the extent to which increasing concentrations of either pyruvate or lactate, compared to glucose, influenced sperm quality during 72-hour storage at 10 or 20 ⁰C was evaluated.
- This experiment also reaffirmed the importance of lower temperature storage (10⁰C), as it resulted in superior sperm quality compared to 20⁰C. Moreover, it was observed that glucose and pyruvate promoted similar sperm motion characteristics and VAI, with reduced VLPP and COMP compared to high concentrations of pyruvate and lactate.
Conclusion
- The presence of glucose in a skim milk-based semen extender optimized stallion sperm quality at storage temperatures of 10⁰C or 20⁰C. However, addition of other substrates such as pyruvate or lactate that readily support oxidative phosphorylation did not improve the sperm quality beyond that provided by glucose alone.
- In fact, these additional substrates seemed to cause deleterious effects on sperm quality over time, apparently due to oxidative stress. Nonetheless, using pyruvate (40 mM) as an energy substrate alternative to glucose offered similar sperm quality results, but only when sperm were stored at 10⁰C.
Cite This Article
APA
Hernández-Avilés C, Ramírez-Agámez L, Love CC, Friedrich M, Pearson M, Kelley DE, Beckham AMN, Teague SR, LaCaze KA, Brinsko SP, Varner DD.
(2020).
The effects of metabolic substrates glucose, pyruvate, and lactate added to a skim milk-based semen extender for cooled storage of stallion sperm.
Theriogenology, 161, 83-97.
https://doi.org/10.1016/j.theriogenology.2020.11.017 Publication
Researcher Affiliations
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, USA. Electronic address: chernandez@cvm.tamu.edu.
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, USA.
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, USA.
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, USA.
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, USA.
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, USA.
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, USA.
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, USA.
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, USA.
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, USA.
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, USA.
MeSH Terms
- Animals
- Glucose / pharmacology
- Horses
- Lactic Acid
- Male
- Milk
- Pyruvic Acid
- Semen
- Semen Preservation / veterinary
- Sperm Motility
- Spermatozoa
Conflict of Interest Statement
Declaration of competing interest There is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported.
Citations
This article has been cited 1 times.- Gacem S, Mocé E, Gozalbo C, Albuixech-Benetó M, Esteve IC, Martínez-Talaván A, Silvestre MA. The Effects of Extender Energetic Substrate Type on Goat Sperm Stored at 17 °C. Biology (Basel) 2025 Jun 27;14(7).
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