Adding cholesterol to the stallion sperm plasma membrane improves cryosurvival.
Abstract: Cryopreservation induces partially irreversible damage to equine sperm membranes. Part of this damage occurs due to membrane alterations induced by the membrane changing from the fluid to the gel-state as the temperature is reduced lower than the membrane transition temperature. One way to prevent this damage is to increase the membrane fluidity at low temperatures by adding cholesterol to the membrane. Different concentrations of cholesterol-loaded-cyclodextrins (CLC) were added to stallion sperm to determine the CLC concentration that optimizes cryosurvival. Higher percentages of motile sperm were maintained after thawing when 1.5 mg CLC was added to sperm from stallions whose sperm do not survive freezing well, compared to control sperm from those same stallions (67% vs. 50%; P<0.05). Addition of CLCs increased the percentages of membrane intact sperm surviving cryopreservation compared to untreated sperm for all stallions (P<0.05). The amount of cholesterol that incorporated into the membranes of the sperm cells increased in a polynomial fashion (R2=0.9978) and incorporated into all sperm membranes. In addition, there was a significant loss of cholesterol from sperm membranes after cryopreservation; however, addition of CLCs to sperm prior to cryopreservation maintained higher cholesterol levels in the sperm after freezing and thawing than untreated sperm (P<0.05). Addition of CLCs also resulted in more sperm binding to the zona pellucida of bovine oocytes after cryopreservation than control sperm (48 vs. 15; P<0.05). In conclusion, CLCs improved the percentage of post-thaw viability in equine sperm as well as increased the number of sperm that bind to zona pellucida. Addition of CLCs to stallion sperm prior to cryopreservation is a simple procedure that increases the cryosurvival of cells.
Publication Date: 2005-08-24 PubMed ID: 16122725DOI: 10.1016/j.cryobiol.2005.07.004Google 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 investigates the effect of adding cholesterol to stallion sperm cells to improve their survival after cryonic freezing. The study found out that adding cholesterol-loaded-cyclodextrins (CLCs) to the cells prior to freezing increases their survival rate and their ability to bind to bovine oocytes after thawing.
Objective of the Research
- The research aimed to mitigate the damage cryopreservation does to equine sperm membranes by increasing their fluidity at low temperatures.
- This was achieved by adding different concentrations of cholesterol-loaded-cyclodextrins (CLCs) to the membranes to find the optimal concentration for cryosurvival.
Methodology and Findings
- CLC was added to stallion sperm cells at different concentrations.
- The researchers found that stallion sperm cells with low cryosurvival rates maintained higher percentages of motile sperm after thawing when 1.5 mg of CLC was added.
- Addition of CLCs also increased the percentages of membrane intact sperm surviving cryopreservation compared to untreated sperm for all stallions.
- There was a significant loss of cholesterol from sperm membranes after cryopreservation, but the addition of CLCs to sperm prior to cryopreservation maintained higher cholesterol levels in the sperm after freezing and thawing.
- In addition, adding CLCs resulted in more sperm binding to the zona pellucida of bovine oocytes after cryopreservation than control sperm.
Conclusion
- The study concludes that adding CLCs to stallion sperm cells prior to cryopreservation improves the percentage of post-thaw viability.
- It also increases the number of sperm cells that can successfully bind to the zona pellucida after thawing.
- Therefore, the simple procedure of adding CLCs to stallion sperm cells prior to cryopreservation has been shown to significantly improve cell cryosurvival.
Cite This Article
APA
Moore AI, Squires EL, Graham JK.
(2005).
Adding cholesterol to the stallion sperm plasma membrane improves cryosurvival.
Cryobiology, 51(3), 241-249.
https://doi.org/10.1016/j.cryobiol.2005.07.004 Publication
Researcher Affiliations
- Department of Biomedical Science, Colorado State University, Fort Collins, CO 80523, USA.
MeSH Terms
- Animals
- Cattle
- Cell Membrane / metabolism
- Cell Survival
- Cholesterol / administration & dosage
- Cholesterol / metabolism
- Cryopreservation / methods
- Cryopreservation / veterinary
- Cryoprotective Agents
- Cyclodextrins
- Female
- Horses
- In Vitro Techniques
- Male
- Membrane Fluidity / drug effects
- Microscopy, Fluorescence
- Semen Preservation / methods
- Semen Preservation / veterinary
- Sperm-Ovum Interactions
- Spermatozoa / cytology
- Spermatozoa / metabolism
- Zona Pellucida / metabolism
Citations
This article has been cited 16 times.- Ligocka Z, Partyka A, Schu00e4fer-Somi S, Mucha A, Niu017cau0144ski W. Does Better Post-Thaw Motility of Dog Sperm Frozen with CLC Mean Better Zona Pellucida Binding Ability?. Animals (Basel) 2023 May 9;13(10).
- Xu X, Hao T, Komba E, Yang B, Hao H, Du W, Zhu H, Zhang H, Zhao X. Improvement of Fertilization Capacity and Developmental Ability of Vitrified Bovine Oocytes by JUNO mRNA Microinjection and Cholesterol-Loaded Methyl-u03b2-Cyclodextrin Treatment.. Int J Mol Sci 2022 Dec 29;24(1).
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