Cryopreservation of epididymal stallion sperm.
Abstract: Any event that makes semen collection or mating impossible, such as death, castration, or injury, may terminate a stallion's breeding career. Fortunately, stallion sperm which are capable of fertilization can be harvested from the epididymis, and frozen for future use. However, the fertility of frozen-thawed epididymal sperm has been found to be lower than that of ejaculated sperm. Therefore, this study aimed to optimize the fertility of frozen epididymal stallion sperm by investigating the effects of different cryoprotectants and freezing protocols on sperm quality. Dimethylformamide was tested alone or combination with pasteurized egg yolk as substitute of fresh egg yolk. In addition, the effect of the pre-freeze stabilization on sperm quality was analyzed. Heterospermic samples obtained from stallion epididymis were collected and cryopreserved in lactose-egg-yolk extender or in the same extender with varying content of cryoprotectant and content of egg yolk, stabilized and no-stabilized. Sperm motility, viability, hypoosmotic swelling test (HOST) and acrosome integrity were evaluated post-thawing. No improvement was observed on the replacement of fresh yolk by pasteurized egg yolk, whereas the results suggest that dimethylformamide is a cryoprotectant suitable for cryopreservation of equine epididymal semen, even better than glycerol. In addition, we found that the stabilization before freezing on epididymal stallion sperm, can improve sperm quality parameters.
Copyright © 2014 Elsevier Inc. All rights reserved.
Publication Date: 2014-01-08 PubMed ID: 24412395DOI: 10.1016/j.cryobiol.2013.12.009Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article investigates how to enhance the fertility of frozen male horse sperm extracted from the epididymis by testing the effects of varied cryoprotectants and freezing methods. The results suggest that using dimethylformamide for cryopreservation and pre-freeze stabilization can improve sperm quality parameters.
Objective of the Research
- The primary goal of this study was to enhance the fertility rates of frozen-thawed stallion sperm extracted from the epididymis. The researchers aimed to attain this by examining the effects of various cryoprotectants and freezing protocols on sperm quality.
Methodology
- Sperm samples were harvested from the epididymis of stallions and frozen with a lactose-egg-yolk extender or with the same extender but with a varied content of cryoprotectant and egg yolk. These samples were stabilized before freezing and also directly frozen without stabilization.
- The sperm quality parameters evaluated in the freeze-thaw cycles included motility, viability, response to hypo-osmotic conditions, and the integrity of the acrosome (the cap-like structure in the head of a sperm).
Findings & Conclusions
- Replacement of fresh egg yolk with pasteurized egg yolk did not yield an improvement in sperm quality. Therefore, this modification was deemed ineffective.
- Using the cryoprotectant dimethylformamide resulted in higher sperm quality compared to the commonly used cryoprotectant, glycerol, making this a promising approach to improving the cryopreservation of equine epididymal sperm.
- Stabilization of sperm before freezing was found to enhance the quality parameters, thus indicating that pre-freeze stabilization may play a key role in improving the fertility potential of frozen-thawed epididymal stallion sperm.
Cite This Article
APA
Olaciregui M, Gil L, Montón A, Luño V, Jerez RA, Martí JI.
(2014).
Cryopreservation of epididymal stallion sperm.
Cryobiology, 68(1), 91-95.
https://doi.org/10.1016/j.cryobiol.2013.12.009 Publication
Researcher Affiliations
- Department of Animal Pathology, Obstetric and Reproduction Area, Faculty of Veterinary Medicine, Universidad de Zaragoza, Spain. Electronic address: maiteor88@gmail.com.
- Department of Animal Pathology, Obstetric and Reproduction Area, Faculty of Veterinary Medicine, Universidad de Zaragoza, Spain.
- Department of Animal Pathology, Obstetric and Reproduction Area, Faculty of Veterinary Medicine, Universidad de Zaragoza, Spain.
- Department of Animal Pathology, Obstetric and Reproduction Area, Faculty of Veterinary Medicine, Universidad de Zaragoza, Spain.
- Department of Animal Pathology, Obstetric and Reproduction Area, Faculty of Veterinary Medicine, Universidad de Zaragoza, Spain.
- Department of Animal Pathology, Obstetric and Reproduction Area, Faculty of Veterinary Medicine, Universidad de Zaragoza, Spain.
MeSH Terms
- Animals
- Cell Survival / drug effects
- Cryopreservation
- Cryoprotective Agents / pharmacology
- Dimethylformamide / pharmacology
- Egg Yolk / chemistry
- Epididymis / cytology
- Epididymis / physiology
- Glycerol / pharmacology
- Horses
- Male
- Osmolar Concentration
- Semen Preservation / methods
- Sperm Motility / drug effects
- Sperm Motility / physiology
- Spermatozoa / drug effects
- Spermatozoa / physiology
Citations
This article has been cited 10 times.- Domingo P, Olaciregui M, González N, De Blas I, Gil L. Effect of glycerol, n, n-dimethylformamide and n-methyl-2-pyrrolidone on rabbit sperm stored at 4 °C and 16 °C. Anim Reprod 2019 Nov 14;16(4):887-894.
- Ryu DY, Song WH, Pang WK, Yoon SJ, Rahman MS, Pang MG. Freezability biomarkers in bull epididymal spermatozoa. Sci Rep 2019 Sep 5;9(1):12797.
- 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.
- Domingo P, Olaciregui M, González N, De Blas I, Gil L. Effects of seminal plasma and different cryoprotectants on rabbit sperm preservation at 16°C. Exp Anim 2018 Nov 1;67(4):413-420.
- Yoon SJ, Rahman MS, Kwon WS, Ryu DY, Park YJ, Pang MG. Proteomic identification of cryostress in epididymal spermatozoa. J Anim Sci Biotechnol 2016;7:67.
- Yoon SJ, Rahman MS, Kwon WS, Park YJ, Pang MG. Addition of Cryoprotectant Significantly Alters the Epididymal Sperm Proteome. PLoS One 2016;11(3):e0152690.
- Patel M, Gandotra VK, Cheema RS, Bansal AK, Kumar A. Seminal Plasma Heparin Binding Proteins Improve Semen Quality by Reducing Oxidative Stress during Cryopreservation of Cattle Bull Semen. Asian-Australas J Anim Sci 2016 Sep;29(9):1247-55.
- Yoon SJ, Kwon WS, Rahman MS, Lee JS, Pang MG. A novel approach to identifying physical markers of cryo-damage in bull spermatozoa. PLoS One 2015;10(5):e0126232.
- Vitoria A, Barrachina L, Romero A, Fuente S, de Blas I, Gil L, Vázquez FJ. Laparoscopic Inguinal Hernioplasty with a Polyether Ether Ketone Anchoring Device in Intact Male Horses Does Not Compromise Testicular Perfusion, Sperm Production or Motility Characteristics. Animals (Basel) 2025 Jan 31;15(3).
- Mofadel HA, Hussein HA, Abd-Elhafee HH, El-Sherry TM. Impact of various cryo-preservation steps on sperm rheotaxis and sperm kinematics in bull. Sci Rep 2024 May 18;14(1):11403.
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