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Effect of cholesterol on the motility and plasma membrane integrity of frozen equine spermatozoa after thawing.
Abstract: The aim of the present study was to investigate the cryoprotectant properties of cholesterol after incorporation into the plasma membranes of equine spermatozoa. A cholesterol-methyl-beta-cyclodextrin complex was used to alter sperm plasma membrane cholesterol content. Ejaculates from six stallions were centrifuged in a non-fat skimmed milk glucose-sucrose extender (MK) or a modified Tyrode's medium (TALP). The sperm pellets were resuspended in the appropriate extender with or without added cholesterol (0.125 mmol cholesterol-methyl-beta-cyclodextrin complex l(-1)) and incubated at 24 degrees C for 15 min. After incubation, the aliquots were centrifuged and the sperm pellets were resuspended in lactose-EDTA-egg yolk freezing extender, frozen in static nitrogen vapour and stored at -196 degrees C. The straws were thawed and the motility and plasma membrane integrity of the spermatozoa were analysed. Addition of cholesterol to the incubation extenders improved the mean percentages of motile, progressively motile and rapidly motile spermatozoa in both the MK and TALP extenders containing cholesterol compared with extenders without cholesterol (P < 0.05). The percentage of spermatozoa with intact plasma membranes was higher in samples incubated in extenders containing cholesterol than in those without cholesterol (P < 0.05). The results of this study indicate that cyclodextrins can be used to incorporate cholesterol into equine sperm plasma membranes and that cholesterol incorporation imparts protection to the spermatozoa during cryopreservation.
Publication Date: 2000-01-01 PubMed ID: 20681124 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
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
This research explored the effects of cholesterol on equine spermatozoa during freezing and thawing processes. The study found that incorporating cholesterol into the spermatozoa’s plasma membranes acted as a protective agent, enhancing sperm motility and membrane integrity post-thawing.
Research Methodology
- The study was designed to explore the cryoprotective properties of incorporating cholesterol into the plasma membranes of equine spermatozoa.
- A cholesterol-methyl-beta-cyclodextrin complex was used to alter the cholesterol content in the sperm plasma membrane.
- Ejaculates obtained from six stallions were prepared for the experiment. They were centrifuged in a non-fat skimmed milk glucose-sucrose extender (MK) or a modified Tyrode’s medium (TALP).
- The centrifuged sperm pellets were then resuspended in the MK or TALP extenders, with or without added cholesterol (0.125 mmol cholesterol-methyl-beta-cyclodextrin complex l(-1)).
- After a 15-minute incubation, the aliquots were centrifuged again, and the sperm pellets were resuspended in a freezing extender made of lactose-EDTA-egg yolk.
- The samples were then frozen using static nitrogen vapour and stored at a temperature of -196 degrees Celsius.
Analysis and Results
- Following the freezing and storage process, the sperm samples were thawed and analysed for sperm motility and plasma membrane integrity.
- The study found that adding cholesterol to the incubation extenders improved the mean percentages of motile, progressively motile, and rapidly motile spermatozoa for both the MK and TALP extenders.
- Moreover, a higher percentage of spermatozoa with intact plasma membranes was observed in samples incubated in extenders containing cholesterol as compared to those without it.
Conclusion
- This research demonstrated the beneficial effects of incorporating cholesterol into equine sperm plasma membranes, discloses that cholesterol incorporation offers protection to the spermatozoa during the freezing and thawing processes.
- This research also proved that cyclodextrins can be effectively used to incorporate cholesterol into equine sperm plasma membranes.
Cite This Article
APA
Combes GB, Varner DD, Schroeder F, Burghardt RC, Blanchard TL.
(2000).
Effect of cholesterol on the motility and plasma membrane integrity of frozen equine spermatozoa after thawing.
J Reprod Fertil Suppl(56), 127-132.
Publication
Researcher Affiliations
- Department of Large Animal Medicine and Surgery, College of Veterinary Medicine, Texas A&M University, College Station, TX 77843-4475, USA.
MeSH Terms
- Animals
- Cell Membrane / drug effects
- Cell Membrane / physiology
- Cholesterol / chemistry
- Cholesterol / pharmacology
- Cryopreservation / veterinary
- Freezing
- Male
- Sperm Motility / drug effects
- Sperm Motility / physiology
- Spermatozoa / cytology
- Spermatozoa / drug effects
- Spermatozoa / physiology
- beta-Cyclodextrins / chemistry
- beta-Cyclodextrins / pharmacology
Citations
This article has been cited 6 times.- Zhang M, Ghosh S, Li M, Altan-Bonnet N, Shuai D. Vesicle-Cloaked Rotavirus Clusters are Environmentally Persistent and Resistant to Free Chlorine Disinfection. Environ Sci Technol 2022 Jun 21;56(12):8475-8484.
- Zhang M, Altan-Bonnet N, Shen Y, Shuai D. Waterborne Human Pathogenic Viruses in Complex Microbial Communities: Environmental Implication on Virus Infectivity, Persistence, and Disinfection. Environ Sci Technol 2022 May 3;56(9):5381-5389.
- Lago-Alvarez Y, Podico G, Segabinazzi LG, Cunha LL, Barbosa L, Arnold CE, Lima FS, King LT, McLean AK, Canisso IF. Donkey Epididymal Transport for Semen Cooling and Freezing. Animals (Basel) 2020 Nov 25;10(12).
- Li S, Ao L, Yan Y, Jiang J, Chen B, Duan Y, Shen F, Chen J, Inglis B, Ni R, Ji W, Si W. Differential motility parameters and identification of proteomic profiles of human sperm cryopreserved with cryostraw and cryovial. Clin Proteomics 2019;16:24.
- Gil L, Galindo-Cardiel I, Malo C, González N, Alvarez C. Effect of Cholesterol and Equex-STM Addition to an Egg Yolk Extender on Pure Spanish Stallion Cryopreserved Sperm. ISRN Vet Sci 2013;2013:280143.
- Kiso WK, Asano A, Travis AJ, Schmitt DL, Brown JL, Pukazhenthi BS. Pretreatment of Asian elephant (Elephas maximus) spermatozoa with cholesterol-loaded cyclodextrins and glycerol addition at 4°C improves cryosurvival. Reprod Fertil Dev 2012;24(8):1134-42.
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