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Detection of “apoptosis-like” changes during the cryopreservation process in equine sperm.
Abstract: The kinematics of the appearance of apoptotic markers was studied by flow cytometry and immunoblot assays in equine spermatozoa subjected to freezing and thawing. Caspase activity, low mitochondrial membrane potential, and increases in sperm membrane permeability were observed in all of the phases of the cryopreservation procedure. Freezing and thawing caused an increase in membrane permeability and changes in the pattern of caspase activity; decreases in mitochondrial membrane potential were observed after centrifugation and cooling to 4 degrees C and after freezing and thawing. It is proposed that sperm mitochondria may be directly involved in the subtle damage that is present in most spermatozoa surviving freezing and thawing.
Publication Date: 2007-10-31 PubMed ID: 17978341DOI: 10.2164/jandrol.107.003640Google 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 looks into how cryopreservation, the process of freezing and thawing, influences changes in the markers of cell death in equine sperm. The study identifies that all stages of cryopreservation lead to signs of cell death including increased caspase enzyme activity, lower energy production, and increased cell membrane permeability.
Overview of Study
- The researchers examined the occurrence and timing of certain indicators of cell death or “apoptosis” in horse sperm during the cryopreservation process. Cryopreservation is a method used to preserve living cells by freezing them at extremely low temperatures.
Methods Used
- The researchers detected these indicators using flow cytometry – a technology that is used to analyse the physical and chemical characteristics of particles in a fluid as it passes through at least one laser – and immunoblot assays, which are used to detect specific proteins in a sample.
Key Findings
- Caspase activity (caspases are a family of protease enzymes that play essential roles in programmed cell death), low mitochondrial membrane potential (a measure of the capacity of the mitochondrial membrane to generate a proton motive force required for ATP production) and increases in sperm membrane permeability were noticed throughout the cryopreservation process.
- Specifically, the freezing and thawing phases led to increases in cell membrane permeability and shifts in caspase activity patterns. The researchers also identified decreases in the potential for energy production in the cells after centrifugation and cooling to 4 degrees celsius and after the freezing and thawing phases.
- Based on these findings, the authors propose that the mitochondria within the sperm cells – the “powerhouse” of the cell that generates energy – may contribute directly to the mild damage found in most of the cells that survive the freezing and thawing process.
Cite This Article
APA
Ortega-Ferrusola C, Sotillo-Galu00e1n Y, Varela-Fernu00e1ndez E, Gallardo-Bolau00f1os JM, Muriel A, Gonzu00e1lez-Fernu00e1ndez L, Tapia JA, Peu00f1a FJ.
(2007).
Detection of “apoptosis-like” changes during the cryopreservation process in equine sperm.
J Androl, 29(2), 213-221.
https://doi.org/10.2164/jandrol.107.003640 Publication
Researcher Affiliations
- Veterinary Teaching Hospital, Laboratory of Spermatology, University of Extremadura, Cu00e1ceres, Spain.
MeSH Terms
- Animals
- Apoptosis
- Benzimidazoles / pharmacology
- Carbocyanines / pharmacology
- Caspases / metabolism
- Cell Membrane Permeability
- Cryopreservation / veterinary
- Flow Cytometry
- Horses
- Male
- Membrane Potential, Mitochondrial
- Semen Preservation / veterinary
- Spermatozoa / cytology
- Spermatozoa / drug effects
Citations
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