Effects of cholesterol-loaded cyclodextrins on the quality of frozen-thawed equine epididymal sperm.
Abstract: Equine epididymal sperm are known to be severely sensitive to cryopreservation, in terms of sperm quality and pregnancy rate. The objective of this study was to examine the effects of cholesterol loaded cyclodextrins (CLCs) on the quality of stallion epididymal sperm during cryopreservation. In experiment I, sperm were treated with different concentrations of CLCs: (1) 0mg (control), (2) 1.5mg, (3) 3mg, and (4) 6 mg per 120 × 10(6) sperm. The sperm viability and amount of cholesterol were determined at 15, 30 and 45 min after CLC treatment using viability markers (Ethidium homodimer-1 and Calcein AM) and gas chromatography, respectively. In experiment II, CLC treated sperm (1.5mg CLC per 120 × 10(6) sperm) were fixed and stained with filipin to examine the cholesterol distribution. In experiment III, sperm were treated with CLCs at concentrations of 1.5, 3.0, 6.0 mg per 120 × 10(6) sperm for 15 min, then equilibrated with freezing extender at 4°C for 1h prior to cryopreservation. Epididymal sperm without CLC loading (0mg) were used as the control group. The sperm quality was examined at post-equilibration and 10 min, 2h and 4h after freezing and thawing. The cholesterol was successfully loaded into the plasma membrane of stallion epididymal sperm. The amount of cholesterol was increased in a manner of dose and time dependence, and the filipin-sterol complexes were increasingly labeled over the sperm head. CLCs at 1.5mg/120 × 10(6) sperm significantly improved sperm quality during sperm equilibration and cryopreservation compared to other doses of CLCs and non-CLC control. An increasing concentration and incubation time of CLCs was detrimental to sperm quality. It is concluded that cholesterol loading to the sperm plasma membrane via CLCs decreases chilling sensitivity and also improves epididymal sperm cryopreservability.
Copyright © 2011 Elsevier Inc. All rights reserved.
Publication Date: 2011-06-12 PubMed ID: 21679702DOI: 10.1016/j.cryobiol.2011.05.006Google 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
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.
The research study investigated how cholesterol-loaded cyclodextrins (CLCs) affect the quality of horse epididymal sperm during freezing and thawing. The main finding suggests that the appropriate application of CLCs could potentially enhance the resilience of this type of sperm against cryopreservation, improving its overall quality.
Research Methodology
- The experiment was carried out in three parts. Initially, epididymal sperm were treated with different concentrations of CLCs and the sperm viability and cholesterol levels were measured at various intervals.
- In the second experiment, CLC treated sperm were fixed and stained to observe the cholesterol distribution.
- In the third part, the sperm were treated with varied concentrations of CLCs before being left to settle with a freezing extender and later cryopreserved. The quality of the sperm was then examined at different checkpoints after freezing and thawing.
Results
- The researchers successfully loaded cholesterol into the sperm plasma membrane and noticed cholesterol levels increased in a dose and time-dependent manner.
- Using a specific dye (filipin), they observed an increase in cholesterol labeling over the sperm head.
- The use of 1.5mg of CLCs per 120 × 10(6) sperm significantly improved sperm quality during equilibration and cryopreservation when compared to other CLC doses and to the control group without CLC loading.
- However, the researchers found that higher concentrations and longer incubation times with CLCs negatively affected sperm quality.
Conclusion
- The study concluded that cholesterol-loading onto the sperm plasma membrane via CLCs not only reduces chilling sensitivity but also improves the freeze-thaw survivability of equine epididymal sperm. These findings may potentially command important implications in animal breeding and veterinary practices, particularly in enhancing the success rate and quality of horse sperm cryopreservation.
Cite This Article
APA
Pamornsakda T, Pojprasath T, Suwimonteerabutr J, Tharasanit T.
(2011).
Effects of cholesterol-loaded cyclodextrins on the quality of frozen-thawed equine epididymal sperm.
Cryobiology, 63(2), 90-95.
https://doi.org/10.1016/j.cryobiol.2011.05.006 Publication
Researcher Affiliations
- Department of Obstetrics, Gynaecology and Reproduction, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330, Thailand.
MeSH Terms
- Animals
- Cell Membrane / metabolism
- Cell Survival
- Cholesterol / metabolism
- Cholesterol / pharmacology
- Cryopreservation / veterinary
- Cryoprotective Agents / pharmacology
- Cyclodextrins / pharmacology
- Epididymis / cytology
- Horses
- Male
- Membrane Fluidity / drug effects
- Semen Analysis / veterinary
- Semen Preservation / veterinary
- Sperm Motility
Citations
This article has been cited 3 times.- 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).
- Ushiyama A, Ishikawa N, Tajima A, Asano A. Comparison of Membrane Characteristics between Freshly Ejaculated and Cryopreserved Sperm in the Chicken.. J Poult Sci 2016 Oct 25;53(4):305-312.
- Behera S, Harshan HM, Bhai KL, Ghosh KN. Effect of cholesterol supplementation on cryosurvival of goat spermatozoa.. Vet World 2015 Dec;8(12):1386-91.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
