Stallion spermatozoa surviving freezing and thawing experience membrane depolarization and increased intracellular Na.
Abstract: In order to gain insight of the modifications that freezing and thawing cause to the surviving population of spermatozoa, changes in the potential of the plasma membrane (Em) and intracellular Na content of stallion spermatozoa were investigated using flow cytometry. Moreover, caspase 3 activity was also investigated and the functionality of the Na -K ATPase pump was investigated before and after freezing and thawing. Cryopreservation caused a significant (p < 0.001) increase in the subpopulation of spermatozoa with depolarized sperm membranes, concomitantly with an increase (p < 0.05) in intracellular Na . These changes occurred in relation to activation of caspase 3 (p < 0.001). Cryopreservation reduced the activity of the Na-K pump and inhibition of the Na -K ATPase pump with ouabain-induced caspase 3 activation. It is concluded that inactivation of Na -K ATPase occurs during cryopreservation, an inhibition that could play a role explaining the accelerated senescence of the surviving population of spermatozoa.
© 2017 American Society of Andrology and European Academy of Andrology.
Publication Date: 2017-10-03 PubMed ID: 28973824DOI: 10.1111/andr.12419Google Scholar: Lookup
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- Journal Article
Summary
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This research investigates the impact of freezing and thawing on stallion sperm cells, showing that this process leads to changes in the cells’ membrane potential and sodium content, which could be linked to an accelerated aging process in the surviving sperm cells.
Objectives and Methodology of the Research
- The goal of this research was to uncover the changes that occur in stallion sperm cells that survive the freeze-thaw process. Specifically, the study sought to find out changes in the potential of the plasma membrane (Em) and intracellular sodium (Na) content.
- The researchers used flow cytometry, a technology that is used to measure the characteristics of single cells in a fluid suspension, to investigate these changes.
- The team also investigated the activity of caspase 3, an enzyme that plays a crucial role in programmed cell death, and the functionality of the sodium-potassium ATPase pump (Na-K pump), both before and after the freezing and thawing process.
Research Findings and Conclusions
- The results showed that cryopreservation, or the process of freezing biological matter at extremely low temperatures, caused a significant increase in the population of sperm cells with depolarized membranes. This depolarization was simultaneously accompanied by an increase in intracellular sodium.
- A key finding was that these changes appeared to be related to the activation of caspase 3. The researchers suggest that this could be a sign of accelerated aging or senescence in the surviving sperm cells.
- Additionally, cryopreservation appeared to reduce the activity of the Na-K pump, which is responsible for maintaining the electrochemical gradient across the cell membrane.
- The researchers concluded that inactivation of the Na-K ATPase pump occurs during cryopreservation. This inhibition could thus be an essential factor in explaining the observed accelerated senescence of the surviving sperm cell population.
Cite This Article
APA
Ortega Ferrusola C, Anel-López L, Ortiz-Rodriguez JM, Martin Muñoz P, Alvarez M, de Paz P, Masot J, Redondo E, Balao da Silva C, Morrell JM, Rodriguez Martinez H, Tapia JA, Gil MC, Anel L, Peña FJ.
(2017).
Stallion spermatozoa surviving freezing and thawing experience membrane depolarization and increased intracellular Na.
Andrology, 5(6), 1174-1182.
https://doi.org/10.1111/andr.12419 Publication
Researcher Affiliations
- Reproduction and Obstetrics Department of Animal Medicine and Surgery, University of León, León, Spain.
- Reproduction and Obstetrics Department of Animal Medicine and Surgery, University of León, León, Spain.
- Laboratory of Equine Reproduction and Equine Spermatology, Veterinary Teaching Hospital, University of Extremadura, Cáceres, Spain.
- Laboratory of Equine Reproduction and Equine Spermatology, Veterinary Teaching Hospital, University of Extremadura, Cáceres, Spain.
- Reproduction and Obstetrics Department of Animal Medicine and Surgery, University of León, León, Spain.
- Department of Molecular Biology, University of León, León, Spain.
- Laboratory of Equine Reproduction and Equine Spermatology, Veterinary Teaching Hospital, University of Extremadura, Cáceres, Spain.
- Laboratory of Equine Reproduction and Equine Spermatology, Veterinary Teaching Hospital, University of Extremadura, Cáceres, Spain.
- Portalagre Polytechnic Institute, Superior Agriculture School of Elvas, Elvas, Portugal.
- Division of Reproduction, Faculty of Veterinary Medicine and Animal Sciences, Swedish University of Agricultural Sciences, Uppsala, Sweden.
- Department of Clinical and Experimental Medicine, Faculty of Medicine & Health Sciences, Linköping University, Linköping, Sweden.
- Laboratory of Equine Reproduction and Equine Spermatology, Veterinary Teaching Hospital, University of Extremadura, Cáceres, Spain.
- Laboratory of Equine Reproduction and Equine Spermatology, Veterinary Teaching Hospital, University of Extremadura, Cáceres, Spain.
- Reproduction and Obstetrics Department of Animal Medicine and Surgery, University of León, León, Spain.
- Laboratory of Equine Reproduction and Equine Spermatology, Veterinary Teaching Hospital, University of Extremadura, Cáceres, Spain.
MeSH Terms
- Animals
- Cell Membrane / pathology
- Cryopreservation / methods
- Freezing
- Horses
- Male
- Semen Preservation / adverse effects
- Semen Preservation / methods
- Sodium / metabolism
- Sodium-Potassium-Exchanging ATPase / metabolism
- Spermatozoa / metabolism
- Spermatozoa / pathology
Citations
This article has been cited 5 times.- Gaitskell-Phillips G, Martín-Cano FE, Ortiz-Rodríguez JM, Silva-Rodríguez A, Gil MC, Ortega-Ferrusola C, Peña FJ. In Stallion Spermatozoa, Superoxide Dismutase (Cu-Zn) (SOD1) and the Aldo-Keto-Reductase Family 1 Member b (AKR1B1) Are the Proteins Most Significantly Reduced by Cryopreservation.. J Proteome Res 2021 May 7;20(5):2435-2446.
- Peña FJ, O'Flaherty C, Ortiz Rodríguez JM, Martín Cano FE, Gaitskell-Phillips GL, Gil MC, Ortega Ferrusola C. Redox Regulation and Oxidative Stress: The Particular Case of the Stallion Spermatozoa.. Antioxidants (Basel) 2019 Nov 19;8(11).
- Ortiz-Rodriguez JM, Balao da Silva C, Masot J, Redondo E, Gazquez A, Tapia JA, Gil C, Ortega-Ferrusola C, Peña FJ. Rosiglitazone in the thawing medium improves mitochondrial function in stallion spermatozoa through regulating Akt phosphorylation and reduction of caspase 3.. PLoS One 2019;14(7):e0211994.
- Ortiz-Rodriguez JM, Ortega-Ferrusola C, Gil MC, Martín-Cano FE, Gaitskell-Phillips G, Rodríguez-Martínez H, Hinrichs K, Álvarez-Barrientos A, Román Á, Peña FJ. Transcriptome analysis reveals that fertilization with cryopreserved sperm downregulates genes relevant for early embryo development in the horse.. PLoS One 2019;14(6):e0213420.
- Ran MX, Li Y, Zhang Y, Liang K, Ren YN, Zhang M, Zhou GB, Zhou YM, Wu K, Wang CD, Huang Y, Luo B, Qazi IH, Zhang HM, Zeng CJ. Transcriptome Sequencing Reveals the Differentially Expressed lncRNAs and mRNAs Involved in Cryoinjuries in Frozen-Thawed Giant Panda (Ailuropoda melanoleuca) Sperm.. Int J Mol Sci 2018 Oct 8;19(10).
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