Post-thaw addition of seminal plasma reduces tyrosine phosphorylation on the surface of cryopreserved equine sperm, but does not reduce lipid peroxidation.
Abstract: The objective was to verify the relationship between equine semen cryopreservation and changes related to increased lipid peroxidation. Also, addition of autologous or homologous seminal plasma from a stallion with a good freezing response to post-thawed sperm was tested to determine whether it would confer protection. Frozen-thawed sperm were evaluated and allocated into three groups: without plasma addition, and supplemented with either homologous or autologous seminal plasma. All groups were evaluated at 0, 60 and 120 min after incubation at 37 °C. Cryopreservation did not increase plasma membrane disorders (mean ± SEM 9.48 ± 0.65 and 1.62 ± 0.23% in raw and frozen-thawed sperm, respectively). However, both membrane peroxidation and protein phosphorylation were increased (P < 0.05) compared to raw semen (1.74 and 5.20-fold, respectively). There was a correlation (r = 0.73; P < 0.05) between the increase in lipid peroxidation and tyrosine phosphorylation. Seminal plasma, regardless of origin, reduced (P > 0.05) tyrosine phosphorylation present on the surface of cryopreserved sperm; however, lipid peroxidation was not significantly reduced. In conclusion, we inferred that emergence of phosphorylated proteins on the surface of cryopreserved sperm was due to increased lipid peroxidation that occurred during the freezing/thawing process; however, reduced tyrosine phosphorylation that occurred after addition of seminal plasma was triggered by other mechanisms, apparently independent from the reduction in membrane peroxidation.
Copyright © 2012 Elsevier Inc. All rights reserved.
Publication Date: 2012-03-22 PubMed ID: 22444550DOI: 10.1016/j.theriogenology.2012.01.003Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research studies how freezing and thawing affects horse semen, specifically focusing on lipid peroxidation and tyrosine phosphorylation. The addition of seminal plasma following thawing was also explored as a potential means of reducing issues, though only a reduction in tyrosine phosphorylation was observed, not lipid peroxidation.
Objectives and Approach
- The research aimed to investigate the effects of cryopreservation on horse semen, with a focus on lipid peroxidation and tyrosine phosphorylation. In particular, it wanted to verify any associated connection between increased lipid peroxidation and the freezing and thawing process.
- Another aim was to observe whether the addition of seminal plasma—sourced from a stallion with a good freezing response—to the post-thawed semen would confer any protective benefits.
- Cryopreserved semen were divided into three groups for comparison: semen with no seminal plasma addition, and semen supplemented with either homologous or autologous seminal plasma.
- The groups were evaluated at different durations (0, 60, and 120 minutes) post-incubation at a regulated temperature.
Findings
- Cryopreservation did not increase plasma membrane disorders, but was seen to cause significant increases in membrane peroxidation and protein phosphorylation compared to unprocessed semen.
- The study found a strong correlation between the proportional increases in lipid peroxidation and tyrosine phosphorylation.
- The addition of seminal plasma (irrespective of its origin) reduced tyrosine phosphorylation on the surface of the cryopreserved sperm, however, it had no significant effect on reducing lipid peroxidation.
Conclusions
- The study concluded that the increased presence of phosphorylated proteins on the surface of cryopreserved sperm was due to increased lipid peroxidation during the freezing/thawing process.
- The reduction in tyrosine phosphorylation observed following the addition of seminal plasma was believed to be caused by mechanisms separate from the reduction in membrane peroxidation.
Cite This Article
APA
de Andrade AF, Zaffalon FG, Celeghini EC, Nascimento J, Bressan FF, Martins SM, de Arruda RP.
(2012).
Post-thaw addition of seminal plasma reduces tyrosine phosphorylation on the surface of cryopreserved equine sperm, but does not reduce lipid peroxidation.
Theriogenology, 77(9), 1866-72.e723.
https://doi.org/10.1016/j.theriogenology.2012.01.003 Publication
Researcher Affiliations
- Department of Animal Reproduction, School of Veterinary Medicine and Animal Science, University of Sao Paulo (USP), Pirassununga, SP, Brazil.
MeSH Terms
- Animals
- Cryopreservation / veterinary
- Flow Cytometry
- Freezing
- Horses / physiology
- Lipid Peroxidation / physiology
- Male
- Phosphorylation
- Semen / physiology
- Semen Preservation / veterinary
- Tyrosine / metabolism
Citations
This article has been cited 7 times.- Ruiz-Díaz S, Oseguera-López I, De La Cuesta-Díaz D, García-López B, Serres C, Sanchez-Calabuig MJ, Gutiérrez-Adán A, Perez-Cerezales S. The Presence of D-Penicillamine during the In Vitro Capacitation of Stallion Spermatozoa Prolongs Hyperactive-Like Motility and Allows for Sperm Selection by Thermotaxis. Animals (Basel) 2020 Aug 21;10(9).
- 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).
- Kumar S, Tinson A, Mulligan BP, Ojha S. Gelatin Binding Proteins in Reproductive Physiology. Indian J Microbiol 2016 Dec;56(4):383-393.
- Torres MA, Díaz R, Boguen R, Martins SM, Ravagnani GM, Leal DF, Oliveira Mde L, Muro BB, Parra BM, Meirelles FV, Papa FO, Dell'Aqua JA Jr, Alvarenga MA, Moretti Ade S, Sepúlveda N, de Andrade AF. Novel Flow Cytometry Analyses of Boar Sperm Viability: Can the Addition of Whole Sperm-Rich Fraction Seminal Plasma to Frozen-Thawed Boar Sperm Affect It?. PLoS One 2016;11(8):e0160988.
- Zambrano F, Pezo F, Furugen Cesar de Andrade A, Rivera-Concha R, Uribe P, Schulz M, Zapparoli H, Mendes de Oliveira Bezerra L, Hermosilla C, Taubert A, Sánchez R. Modulation of NETosis in Swine Neutrophil-Spermatozoa Co-Cultures In Vitro: Effects of Butylated Hydroxytoluene, Albumin, Prostaglandin E(2), and Seminal Plasma. Antioxidants (Basel) 2025 Jun 24;14(7).
- Wen X, Bou G, He Q, Liu Q, Yi M, Ren H. Comprehensive Integrated Analyses of Proteins and Metabolites in Equine Seminal Plasma (Horses and Donkeys). Proteomes 2025 Jul 4;13(3).
- Ledesma A, Zalazar L, Buchelly Imbachi F, Pastore JI, Brown P, Eddy EM, Hozbor F, Cesari A. Recombinant peptide reverses cryo-capacitation in ram sperm and improves in vitro fertilization. Anim Reprod Sci 2019 Aug;207:61-72.
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