Sex sorting increases the permeability of the membrane of stallion spermatozoa.
Abstract: At present, the only repeatable means of selecting the sex of offspring is the Beltsville semen sorting technology using flow cytometry (FC). This technology has reached commercial status in the bovine industry and substantial advances have occurred recently in swine and ovine species. In the equine species, however, the technology is not as well developed. To better understand the changes induced in stallion spermatozoa during the sorting procedure, pooled sperm samples were sorted: sperm motility and kinematics were assessed using computer assisted sperm analysis, sperm membrane integrity was assessed using the YoPro-1 assay, while plasmalemmal stability and lipid architecture were assessed using Merocyanine 540/SYTOX green and Annexin-V, respectively. Lipid peroxidation was also investigated with the probe Bodipy(581/591)-C11. All assays were performed shortly after collection, after incubation and after sex sorting using FC. In order to characterize potential molecular mechanisms implicated in sperm damage, an apoptosis protein antibody dot plot array analysis was performed before and after sorting. While the percentage of total motile sperm remained unchanged, sex sorting reduced the percentages of progressive motile spermatozoa and of rapid spermatozoa as well as curvilinear velocity (VCL). Sperm membranes responded to sorting with an increase in the percentage of YoPro-1 positive cells, suggesting the sorted spermatozoa had a reduced energy status that was confirmed by measuring intracellular ATP content.
Copyright © 2013 Elsevier B.V. All rights reserved.
Publication Date: 2013-03-14 PubMed ID: 23567220DOI: 10.1016/j.anireprosci.2013.02.021Google 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.
- Evaluation Study
- 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 investigates the impact of sex sorting on the permeability of the membrane of stallion spermatozoa. The findings suggest that while sorting doesn’t significantly impact overall sperm mobility, it has negative effects on the progression and speed of the sperm and potentially reduces their energy status.
Context
- This study focuses on the use of Beltsville semen sorting technology, which has been successfully commercialized in the bovine industry and shows promise in other livestock species. However, its application in equine species is not yet well-developed.
- The researchers sought to understand how stallion spermatozoa change throughout the sorting procedure.
Methodology
- Pooled sperm samples were sorted using flow cytometry (FC), a technique used to assess the physical and chemical characteristics of particles in a fluid as they pass through at least one laser.
- The researchers studied sperm motility and kinematics through computer-assisted sperm analysis and assessed the sperm membrane integrity using the YoPro-1 assay.
- Lipid peroxidation was also investigated while plasmalemmal stability and lipid architecture were tested using Merocyanine 540/SYTOX green and Annexin-V, respectively.
Findings
- The results showed that despite sex sorting not impacting the total percentage of motile sperm, it reduced the percentage of progressive motile spermatozoa and rapid spermatozoa along with curvilinear velocity (VCL), which is the speed along the curve that the sperm moves.
- Furthermore, the sperm responded to sorting with an increase in the percentage of YoPro-1 positive cells, which indicates a higher membrane permeability, hinting that the sorted sperm had a reduced energy status. This was also proven by measuring the intracellular ATP content, which indicates the energy available to the sperm.
- The researchers also investigated potential molecular mechanisms that could explain the sperm damage, using an apoptosis protein antibody dot plot array analysis before and after sorting.
Cite This Article
APA
Balao da Silva CM, Ortega Ferrusola C, Morillo Rodriguez A, Gallardo Bolaños JM, Plaza Dávila M, Morrell JM, Rodriguez Martínez H, Tapia JA, Aparicio IM, Peña FJ.
(2013).
Sex sorting increases the permeability of the membrane of stallion spermatozoa.
Anim Reprod Sci, 138(3-4), 241-251.
https://doi.org/10.1016/j.anireprosci.2013.02.021 Publication
Researcher Affiliations
- Laboratory of Equine Reproduction and Equine Spermatology, Veterinary Teaching Hospital.
MeSH Terms
- Adenosine Triphosphate / metabolism
- Animals
- Calcium / metabolism
- Cell Membrane Permeability
- Cell Separation / methods
- Cell Separation / veterinary
- Cell Survival / physiology
- Female
- Flow Cytometry / methods
- Flow Cytometry / veterinary
- Horses
- Lipid Peroxidation / physiology
- Male
- Phosphatidylserines / metabolism
- Sex Preselection / veterinary
- Sperm Motility
- Spermatozoa / metabolism
Citations
This article has been cited 9 times.- Reese S, Pirez MC, Steele H, Kölle S. The reproductive success of bovine sperm after sex-sorting: a meta-analysis. Sci Rep 2021 Aug 30;11(1):17366.
- Steele H, Makri D, Maalouf WE, Reese S, Kölle S. Bovine Sperm Sexing Alters Sperm Morphokinetics and Subsequent Early Embryonic Development. Sci Rep 2020 Apr 10;10(1):6255.
- Peña FJ, O'Flaherty C, Ortiz Rodríguez JM, Martín Cano FE, Gaitskell-Phillips GL, Gil MC, Ortega Ferrusola C. Redox Regulation and Oxidative Stress: The Particular Case of the Stallion Spermatozoa. Antioxidants (Basel) 2019 Nov 19;8(11).
- Saalfeld GQ, Varela Junior AS, Castro T, Pires DM, Pereira JR, Pereira FA, Corcini CD, Colares EP. Atrazine exposure in gestation and breastfeeding affects Calomys laucha sperm cells. Environ Sci Pollut Res Int 2019 Dec;26(34):34953-34963.
- Daloglu MU, Lin F, Chong B, Chien D, Veli M, Luo W, Ozcan A. 3D imaging of sex-sorted bovine spermatozoon locomotion, head spin and flagellum beating. Sci Rep 2018 Oct 23;8(1):15650.
- Saalfeld GQ, Varela Junior AS, Castro T, Pereira FA, Gheller SMM, da Silva AC, Corcini CD, da Rosa CE, Colares EP. Low atrazine dosages reduce sperm quality of Calomys laucha mice. Environ Sci Pollut Res Int 2018 Jan;25(3):2924-2931.
- Umezu K, Hiradate Y, Numabe T, Hara K, Tanemura K. Effects on glycocalyx structures of frozen-thawed bovine sperm induced by flow cytometry and artificial capacitation. J Reprod Dev 2017 Oct 18;63(5):473-480.
- Kalo D, Manovich S, Yaacobi-Artzi S, Roth Z. Association between sex and the developmental morphokinetics of in vitro derived bovine embryos. Sci Rep 2025 Aug 5;15(1):28631.
- Kalo D, Yaacobi-Artzi S, Manovich S, Michaelov A, Komsky-Elbaz A, Roth Z. Environmental Stress-Induced Alterations in Embryo Developmental Morphokinetics. J Xenobiot 2024 Oct 21;14(4):1613-1637.
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
