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Cryotolerance of stallion spermatozoa is related to ROS production and mitochondrial membrane potential rather than to the integrity of sperm nucleus.
Abstract: Although cryopreservation of stallion spermatozoa allows long-term preservation of spermatozoa from particular stallions and facilitates international trade, it is understood to inflict damages on sperm cells that may finally reduce their fertilizing ability. In addition, individual differences are known to exist in the sperm ability to withstand freeze-thawing protocols. To date, these differences have mainly been reported on the basis of sperm motility and membrane integrity. For this reason, the present work sought to determine differences between good (good freezability ejaculates: GFE) and poor (poor freezability ejaculates: PFE) freezability stallion ejaculates in other sperm parameters, including peroxide and superoxide levels, potential of mitochondrial membrane and nuclear integrity. With this purpose, a total of 24 stallion ejaculates were cryopreserved and classified into two groups (GFE vs. PFE), depending on their sperm membrane integrity and motility after freeze-thawing. From the total of 24 ejaculates, 13 were classified as GFE and the other 11 were classified as PFE. Apart from differences in sperm membrane permeability and lipid disorder after freeze-thawing, GFE presented significantly (p < 0.05) higher percentages of viable spermatozoa with high content of peroxides and of superoxides than PFE. In contrast, and despite cryopreservation of stallion spermatozoa increasing DNA fragmentation and disrupting disulphide bonds in sperm head proteins, no significant differences between GFE and PFE were seen. We can thus conclude that good and poor freezability stallion ejaculates differ in their reactive oxygen species levels after cryopreservation, but not in the damage extent on sperm nucleus.
© 2014 American Society of Andrology and European Academy of Andrology.
Publication Date: 2014-10-08 PubMed ID: 25294093DOI: 10.1111/andr.291Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article investigates the survival factors of horse sperm during freeze-thawing (cryopreservation) protocols. Results show that cryotolerance is more reliant on reactive oxygen species (ROS) production and mitochondrial membrane potential, rather than the integrity of the sperm’s nucleus.
Introduction to the Study
- The study was driven by the established fact that cryopreservation causes substantial damage to sperm cells, which negatively impacts their fertilization ability.
- There are also known individual variances in how well different sperms endure freeze-thawing processes.
- Prior research has primarily reported these differences based on sperm motility and membrane integrity, however, this study explores more parameters, such as peroxide and superoxide levels, mitochondrial membrane potential, and nuclear integrity.
Methodology
- 24 stallion ejaculates were collected and cryopreserved for the purpose of this study.
- The samples were divided into two groups based on their sperm membrane integrity and motility after undergoing the freezing and thawing process: Good Freezability Ejaculates (GFE) and Poor Freezability Ejaculates (PFE).
- There were 13 GFE and 11 PFE from the total 24 ejaculates.
Findings
- Analysis showed that apart from differences in sperm membrane permeability and lipid disorder after freeze-thawing, GFE presented significantly higher percentages of viable sperm with high peroxide and superoxide content compared to PFE.
- Contrarily, despite the cryopreservation process increasing DNA fragmentation and disrupting disulphide bonds in sperm head proteins, no significant differences were observed between GFE and PFE in respect to the damage extent to sperm nucleus.
Conclusion
- At the end of the study, the researchers concluded the ability of horse sperm to survive freeze-thaw protocols is driven more by the levels of oxidative stress (reactive oxygen species) after cryopreservation, not the extent of damage inflicted on the sperm’s nucleus.
Cite This Article
APA
Yeste M, Estrada E, Rocha LG, Maru00edn H, Rodru00edguez-Gil JE, Miru00f3 J.
(2014).
Cryotolerance of stallion spermatozoa is related to ROS production and mitochondrial membrane potential rather than to the integrity of sperm nucleus.
Andrology, 3(2), 395-407.
https://doi.org/10.1111/andr.291 Publication
Researcher Affiliations
- Unit of Animal Reproduction, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Autonomous University of Barcelona, Bellaterra, Barcelona, Spain.
MeSH Terms
- Adaptation, Physiological
- Animals
- Cell Nucleus
- Cryopreservation
- Horses
- Male
- Membrane Potential, Mitochondrial
- Reactive Oxygen Species / metabolism
- Semen Preservation
- Spermatozoa / metabolism
- Spermatozoa / physiology
Citations
This article has been cited 18 times.- Li S, Ren J, Zhang W, Wang B, Ma Y, Su L, Dai Y, Liu G. Glutathione and selenium nanoparticles have a synergistic protective effect during cryopreservation of bull semen.. Front Vet Sci 2023;10:1093274.
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- Tanpradit N, Chatdarong K, Comizzoli P. Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP) pre-exposure ensures follicle integrity during in vitro culture of ovarian tissue but not during cryopreservation in the domestic cat model.. J Assist Reprod Genet 2016 Dec;33(12):1621-1631.
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