L-carnitine and pyruvate are prosurvival factors during the storage of stallion spermatozoa at room temperature.
Abstract: The spermatozoa of many stallions do not tolerate being cooled, restricting the commercial viability of these animals and necessitating the development of a chemically defined room temperature (RT) storage medium. This study examined the impact of two major modulators of oxidative phosphorylation, pyruvate (Pyr) and L-carnitine (L-C), on the storage of stallion spermatozoa at RT. Optimal concentrations of Pyr (10 mM) and L-C (50 mM) were first identified and these concentrations were then used to investigate the effects of these compounds on sperm functionality and oxidative stress at RT. Mitochondrial and cytosolic reactive oxygen species, along with lipid peroxidation, were all significantly suppressed by the addition of L-C (48 h MitoSOX Red negative: 46.2% vs. 26.1%; 48 and 72 h dihydroethidium negative: 61.6% vs. 43.1% and 64.4% vs. 46.9%, respectively; 48 and 72 h 4-hydroxynonenal negative: 37.1% vs. 23.8% and 41.6% vs. 25.7%, respectively), while the Pyr + L-C combination resulted in significantly higher motility compared to the control at 72 h (total motility: 64.2% vs. 39.4%; progressive motility: 34.2% vs. 15.2%). In addition, supplementation with L-C significantly reduced oxidative DNA damage at 72 h (9.0% vs. 15.6%). To investigate the effects of L-C as an osmolyte, comparisons were made between media that were osmotically balanced with NaCl, choline chloride, or L-C. This analysis demonstrated that spermatozoa stored in the L-C balanced medium had significantly higher total motility (55.0% vs. 39.0%), rapid motility (44.0% vs. 25.7%), and ATP levels (70.9 vs. 12.8 ng/ml) following storage compared with the NaCl treatment, while choline chloride did not significantly improve these parameters compared to the control. Finally, mass spectrometry was used to demonstrate that a combination of Pyr and L-C produced significantly higher acetyl-L-carnitine production than any other treatment (6.7 pg/10(6) spermatozoa vs. control at 4.0 pg/10(6) spermatozoa). These findings suggest that Pyr and L-C could form the basis of a novel, effective RT storage medium for equine spermatozoa.
© 2015 by the Society for the Study of Reproduction, Inc.
Publication Date: 2015-08-26 PubMed ID: 26316064DOI: 10.1095/biolreprod.115.131326Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research investigates how the compounds pyruvate and L-carnitine can improve the survival of stallion sperm when stored at room temperature. The study shows that these compounds enhance sperm functionality and reduce oxidative stress, suggesting they could be used to develop a new storage medium for equine sperm.
Introduction
- The main focus of the research is on the storage of stallion spermatozoa at room temperature using pyruvate and L-carnitine. This is important as many stallions’ sperm do not fare well under cooling, posing limitations to their commercial viability in breeding and necessitating alternative storage methods.
Optimal Concentrations and Functionality
- The study began by determining the optimal concentrations of pyruvate and L-Carnitine to use for this process, which were found to be 10 mM and 50 mM, respectively.
- These concentrations were then used to explore the effects of the compounds on sperm functionality and the levels of oxidative stress during storage.
- Results showed that L-carnitine significantly suppressed both mitochondrial and cytosolic reactive oxygen species, as well as lipid peroxidation, all of which are damaging to cell function.
- A combination of pyruvate and L-carnitine resulted in significantly higher motility compared to the control group at 72 hours, and L-Carnitine alone reduced oxidative DNA damage.
Osmolyte Effects
- The researchers also studied the effects of L-Carnitine as an osmolyte, comparing its performance to sodium chloride (NaCl) and choline chloride.
- Spermatozoa in the L-Carnitine balanced medium demonstrated significantly higher total and rapid motility, as well as ATP levels, in comparison with those in the NaCl treatment, whereas choline chloride did not significantly improve these parameters compared to the control group.
Mass Spectrometry and Conclusion
- The team used mass spectrometry to confirm that a combination of pyruvate and L-carnitine led to a significantly higher production of acetyl-L-carnitine (an important metabolite) compared with other treatments.
- The study concludes that pyruvate and L-carnitine could form the basis of an effective new room temperature storage medium for equine spermatozoa.
Cite This Article
APA
Gibb Z, Lambourne SR, Quadrelli J, Smith ND, Aitken RJ.
(2015).
L-carnitine and pyruvate are prosurvival factors during the storage of stallion spermatozoa at room temperature.
Biol Reprod, 93(4), 104.
https://doi.org/10.1095/biolreprod.115.131326 Publication
Researcher Affiliations
- Priority Research Centre for Reproductive Science, Discipline of Biological Sciences, Faculty of Science and IT, University of Newcastle, Callaghan, New South Wales, Australia zamira.gibb@newcastle.edu.au.
- Priority Research Centre for Reproductive Science, Discipline of Biological Sciences, Faculty of Science and IT, University of Newcastle, Callaghan, New South Wales, Australia.
- Priority Research Centre for Reproductive Science, Discipline of Biological Sciences, Faculty of Science and IT, University of Newcastle, Callaghan, New South Wales, Australia.
- Analytical and Biomolecular Research Facility, Central Scientific Services, Research Services, University of Newcastle, Callaghan, New South Wales, Australia.
- Priority Research Centre for Reproductive Science, Discipline of Biological Sciences, Faculty of Science and IT, University of Newcastle, Callaghan, New South Wales, Australia.
MeSH Terms
- Acrosome / drug effects
- Adenosine Triphosphate / metabolism
- Animals
- Carnitine / pharmacology
- Choline / pharmacology
- Chromatin / drug effects
- Chromatin / ultrastructure
- DNA Damage
- Horses
- Lipid Peroxidation / drug effects
- Male
- Mitochondria / drug effects
- Mitochondria / ultrastructure
- Osmolar Concentration
- Pyruvic Acid / pharmacology
- Reactive Oxygen Species / metabolism
- Semen Preservation
- Sperm Motility / drug effects
- Spermatozoa / drug effects
- Spermatozoa / metabolism
- Temperature
Citations
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