Influence of spermatozoal lipidomic profile on the cryoresistance of frozen spermatozoa from stallions.
Abstract: The membrane of spermatozoa, which contributes to cellular cryoresistance, contains numerous lipids with a composition that directly affects membrane fluidity and the fertilization process. In light of variations in the degree of sensitivity in equine seminal freezing, this study aimed to correlate equine semen lipids with post-thawing characteristics of spermatozoa. We used ejaculates from 34 stallions, which were evaluated (total motility ≥ 60%), frozen and thawed and reevaluated for motility of spermatozoa, membrane integrity and lipid peroxidation. Lipid extraction of the fresh semen samples was performed by liquid-liquid extraction, and fingerprinting lipid analysis was conducted by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Based on the characteristics of spermatozoa after thawing, the animals could be separated into two groups: resistant (Good Freezers, n = 5) and sensitive (Bad Freezers, n = 6) to freezing, and their MALDI-MS data were then compared. The Good Freezers group showed a higher abundance of phosphatidylcholines (m/z 796.6, 846.6, 810.6, 854.6 and 732.6). The ions of m/z 812.6, 832.6, 836.6 and 838.6 belonging to the phosphatidylcholine lipid class were also positively correlated with motility of spermatozoa, whereas that of m/z 794.6 was negatively correlated with lipid peroxidation in thawed semen. The Bad Freezer group, displayed higher abundance of one phosphatidylcholines (m/z 806.6), as well as a sphingomyelins (m/z 703.5), which were negatively correlated (univariate analysis) with kinetics of spermatozoa after thawing (m/z 703.5) and with membrane integrity (m/z 792.6). The ion of m/z 717.5, assigned to phosphatidic acid, was negatively correlated with lipid peroxidation. In general therefore, the phosphatidylcholines are associated with higher quality of spermatozoa after thawing, especially in functional capacity, and that lipid semen composition was found to influence the resistance of spermatozoa to cryopreservation and may interfere with motility, membrane integrity and lipid peroxidation in stallions.
Copyright © 2017 Elsevier Inc. All rights reserved.
Publication Date: 2017-11-26 PubMed ID: 29223009DOI: 10.1016/j.theriogenology.2017.11.025Google Scholar: Lookup
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- Journal Article
Summary
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The research article focuses on analyzing the link between the lipid content in horse sperm and their resistance to freezing and thawing processes. The study identifies certain lipid profiles associated with strong post-thaw sperm characteristics that could influence the preservation and overall functionality of horse semen.
Research Objective
- The main purpose of this study was to examine the correlation between the lipid composition of equine semen and the post-thaw characteristics of sperm. The researchers attempted to understand why some horse sperm samples weather the freezing process better than others by studying these samples’ lipidomic profiles.
Methodology
- The study was conducted using the ejaculates from 34 stallions, all of which were first tested for total motility before being frozen and thawed. After the thawing process, the sperm samples were reevaluated for motility, membrane integrity, and lipid peroxidation.
- The lipid content of the fresh semen samples was extracted using a liquid-liquid extraction method and fingerprinting lipid analysis was conducted using a technique called matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS).
Results
- Based on the post-thaw characteristics, the sperm samples were categorized into two groups – ‘Good Freezers’ and ‘Bad Freezers’ – and their corresponding MALDI-MS data were compared. ‘Good Freezers’ were those with more resilient characteristics after thawing, while ‘Bad Freezers’ demonstrated sensitivity to the freezing process.
- The ‘Good Freezers’ showed a higher proportion of phosphatidylcholines in their lipidomic profile. Certain phosphatidylcholine profiles were also positively correlated with the motility of sperm, while one specific profile was negatively correlated with lipid peroxidation in thawed semen.
- The ‘Bad Freezers’ had higher proportions of different phosphatidylcholines and a lipid known as sphingomyelins in their lipid profile. These specific lipids were negatively correlated with the kinetics of sperm post-thaw and also with membrane integrity.
- Another lipid, phosphatidic acid, was found to negatively correlate with lipid peroxidation.
Conclusions
- The study concluded that the lipid profile, specifically the presence of phosphatidylcholines, in horse semen was a significant influencing factor in determining the resilience of sperm to freezing and thawing. The lipid composition could affect motility, membrane integrity, and lipid peroxidation post-thaw.
- This information could potentially help improve methods of semen preservation, ensuring better survival and functionality after thawing.
Cite This Article
APA
Cabrera T, Ramires-Neto C, Belaz KRA, Freitas-Dell'aqua CP, Zampieri D, Tata A, Eberlin MN, Alvarenga MA, Souza FF.
(2017).
Influence of spermatozoal lipidomic profile on the cryoresistance of frozen spermatozoa from stallions.
Theriogenology, 108, 161-166.
https://doi.org/10.1016/j.theriogenology.2017.11.025 Publication
Researcher Affiliations
- Department of Animal Reproduction and Veterinary Radiology, Faculty of Veterinary Medicine, FMVZ, São Paulo State University (UNESP), Botucatu, Brazil.
- Department of Animal Reproduction and Veterinary Radiology, Faculty of Veterinary Medicine, FMVZ, São Paulo State University (UNESP), Botucatu, Brazil.
- ThoMSon Mass Spectrometry Laboratory, Institute of Chemistry, University of Campinas (UNICAMP), Campinas, SP, Brazil; Institute of Chemistry, Federal University of Uberlândia (UFU), Uberlândia, MG, Brazil.
- Department of Animal Reproduction and Veterinary Radiology, Faculty of Veterinary Medicine, FMVZ, São Paulo State University (UNESP), Botucatu, Brazil.
- ThoMSon Mass Spectrometry Laboratory, Institute of Chemistry, University of Campinas (UNICAMP), Campinas, SP, Brazil.
- ThoMSon Mass Spectrometry Laboratory, Institute of Chemistry, University of Campinas (UNICAMP), Campinas, SP, Brazil.
- ThoMSon Mass Spectrometry Laboratory, Institute of Chemistry, University of Campinas (UNICAMP), Campinas, SP, Brazil.
- Department of Animal Reproduction and Veterinary Radiology, Faculty of Veterinary Medicine, FMVZ, São Paulo State University (UNESP), Botucatu, Brazil.
- Department of Animal Reproduction and Veterinary Radiology, Faculty of Veterinary Medicine, FMVZ, São Paulo State University (UNESP), Botucatu, Brazil. Electronic address: fafesouza@fmvz.unesp.br.
MeSH Terms
- Animals
- Cryopreservation / veterinary
- Freezing
- Horses / physiology
- Lipid Metabolism
- Lipids / classification
- Male
- Semen
- Semen Analysis / veterinary
- Semen Preservation / veterinary
- Sperm Motility
- Spermatozoa / physiology
Citations
This article has been cited 8 times.- An Z, Xun W, Zhou H, Hou G, Shi L. Effectiveness of Curcumin on Oxidative Stress in Goat Semen: Explorations Regarding Semen Quality, Sperm Apoptosis, Ultrastructure, and Markers of Oxidative Stress. Antioxidants (Basel) 2025 Oct 16;14(10).
- Castro M, Leal K, Pezo F, Contreras MJ. Sperm Membrane: Molecular Implications and Strategies for Cryopreservation in Productive Species. Animals (Basel) 2025 Jun 19;15(12).
- An Z, Shi L, Zhou H, Hou G, Xun W. Exploratory Metabolomics and Lipidomics Profiling Contributes to Understanding How Curcumin Improves Quality of Goat Semen Stored at 16 °C in Tropical Areas. Int J Mol Sci 2024 Sep 23;25(18).
- Li X, Guo Y, Wang X, Li H, Mao J, Yan S, Luo G, Wang R, Wu X, Li Y. Seminal plasma metabolomics signatures of normosmic congenital hypogonadotropic hypogonadism. Heliyon 2023 Apr;9(4):e14779.
- Harris IT, Maddock C, Farnworth M, Nankervis K, Perrett J, Pyatt AZ, Blanchard RN. Temporal trends in equine sperm progressive motility: a systematic review and meta-regression. Reproduction 2023 Jun 1;165(6):M1-M10.
- Xu B, Bai X, Zhang J, Li B, Zhang Y, Su R, Wang R, Wang Z, Lv Q, Zhang J, Li J. Metabolomic analysis of seminal plasma to identify goat semen freezability markers. Front Vet Sci 2023;10:1132373.
- Xu B, Wang R, Wang Z, Liu H, Wang Z, Zhang W, Zhang Y, Su R, Liu Z, Liu Y, Li J, Zhang J. Evaluation of lipidomic change in goat sperm after cryopreservation. Front Vet Sci 2022;9:1004683.
- Šichtař J, Bubeníčková F, Sirohi J, Šimoník O. How to Increase Post-Thaw Semen Quality in Poor Freezing Stallions: Preliminary Results of the Promising Role of Seminal Plasma Added after Thawing. Animals (Basel) 2019 Jul 3;9(7).
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