Impact of post-thaw supplementation of semen extender with antioxidants on the quality and function variables of stallion spermatozoa.
Abstract: During cryopreservation procedures, the spermatozoa are exposed to physical and chemical stressors that generate an increase in the intracellular concentration of reactive oxygen species (ROS). If ROS concentrations are too great, this can lead to a state of oxidative stress that are detrimental to sperm quality. The aim of this study was to ascertain the profile the ROS production and assess the effects of post-thaw supplementation of a semen extender with different antioxidant compounds on the quality and function variables of frozen-thawed stallion spermatozoa incubated in vitro. Frozen-thawed stallion spermatozoa (2 × 10 cells/mL) were incubated with three different antioxidants (MnTBAP, NAC and FeTPPS) for 4 h at 38 °C. An untreated sperm suspension and a fresh sample were included as controls. Plasma membrane integrity (SYBR-14/PI), intracellular ROS concentration (DHE and ROS-ID™ total ROS/Superoxide Detection Kit), lipid peroxidation (BODIPY), DNA damage (TUNEL) and mitochondrial membrane potential (ΔΨm; TMRE/SYTOX) were evaluated by flow cytometry and fluorescence microscopy. In addition, sperm motility was evaluated using the ISAS system. Evaluations were performed at 0 and 4 h of incubation. The results indicate that superoxide anion is the main ROS produced by frozen-thawed stallion spermatozoa and that the use of MnTBAP improved sperm motility and viability, decreased the lipid peroxidation and DNA damage. In conclusion, this study provides relevant data to improve in vitro incubations conditions and to establish futures therapies using MnTBAP after thawing with the aim being to overcome the deleterious effects of semen cryopreservation and consequently preserve the stallion sperm quality through avoiding oxidative stress.
Copyright © 2018 Elsevier B.V. All rights reserved.
Publication Date: 2018-12-20 PubMed ID: 30591234DOI: 10.1016/j.anireprosci.2018.12.011Google Scholar: Lookup
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- Journal Article
Summary
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The research article examines how adding antioxidants to a semen extender—used after the freeze-thaw cycle of sperm preservation—can improve the quality of stallion sperm by reducing oxidative stress, which is typically harmful to sperm quality.
Introduction and Aims
- The study aims to understand the effects of adding three different antioxidant compounds to a semen extender on the quality and function of cryopreserved (frozen and then thawed) stallion spermatozoa that are incubated in vitro.
- The need for this study arises from the fact that during cryopreservation, sperm cells are exposed to physical and chemical stressors which lead to an increase in reactive oxygen species (ROS). High concentrations of ROS in the cells can cause oxidative stress which is damaging to sperm quality.
Methodology
- Specifically, the study looks at the effects of adding three antioxidants (MnTBAP, NAC, and FeTPPS) to a semen extender in which cryopreserved stallion sperm is incubated.
- Various sperm quality parameters such as viability, ROS concentration, lipid peroxidation, DNA damage, and mitochondrial membrane potential were evaluated using various techniques like flow cytometry, fluorescence microscopy and the ISAS system. Evaluations were performed both at the start and after 4 hours of incubation.
- An untreated sperm suspension and a fresh sample were used as controls to compare the results.
Findings
- The study primarily found that superoxide anion was the main ROS produced by thawed stallion spermatozoa.
- Among the three antioxidants, the use of MnTBAP showed significant improvements in the sperm health. Specifically, it improved sperm motility and viability, and decreased both the degree of lipid peroxidation and DNA damage.
Conclusion
- In conclusion, this study provides important insights that could improve in vitro incubation conditions. Going forward, using MnTBAP after thawing could be an effective strategy to overcome the harmful effects of semen cryopreservation and maintain sperm quality.
Cite This Article
APA
Treulen F, Aguila L, Arias ME, Jofré I, Felmer R.
(2018).
Impact of post-thaw supplementation of semen extender with antioxidants on the quality and function variables of stallion spermatozoa.
Anim Reprod Sci, 201, 71-83.
https://doi.org/10.1016/j.anireprosci.2018.12.011 Publication
Researcher Affiliations
- Laboratory of Reproduction, Centre of Reproductive Biotechnology (CEBIOR-BIOREN), Faculty of Medicine, Universidad de La Frontera, Temuco, Chile; Department of Agricultural Sciences and Natural Resources, Faculty of Agriculture and Forestry, Universidad de La Frontera, Temuco, Chile; School of Medical Technology, Faculty of Science, Universidad Mayor, Temuco, Chile.
- Centre de recherche en reproduction et fértilité (CRRF), Université de Montréal, St-Hyacinthe, QC, Canada.
- Laboratory of Reproduction, Centre of Reproductive Biotechnology (CEBIOR-BIOREN), Faculty of Medicine, Universidad de La Frontera, Temuco, Chile; Department of Animal Production, Faculty of Agriculture and Forestry Sciences, Universidad de La Frontera, Temuco, Chile.
- Laboratory of Neurosciences and Biological Peptides, Centre of Reproductive Biotechnology (CEBIOR-BIOREN), Faculty of Medicine. Universidad de La Frontera, Chile.
- Laboratory of Reproduction, Centre of Reproductive Biotechnology (CEBIOR-BIOREN), Faculty of Medicine, Universidad de La Frontera, Temuco, Chile; Department of Agricultural Sciences and Natural Resources, Faculty of Agriculture and Forestry, Universidad de La Frontera, Temuco, Chile. Electronic address: rfelmerd@gmail.com.
MeSH Terms
- Animals
- Antioxidants / pharmacology
- Cell Membrane
- Cryopreservation / veterinary
- DNA Fragmentation
- Horses / physiology
- Lipid Peroxidation
- Male
- Membrane Potential, Mitochondrial
- Reactive Oxygen Species
- Semen Analysis / veterinary
- Semen Preservation / methods
- Semen Preservation / veterinary
- Spermatozoa / drug effects
Citations
This article has been cited 5 times.- Levano G, Quispe J, Vargas D, García M, López A, Aguila L, Valdivia M. Effect of Atomized Black Maca (Lepidium meyenii) Supplementation in the Cryopreservation of Alpaca (Vicugna pacos) Epididymal Spermatozoa.. Animals (Basel) 2023 Jun 21;13(13).
- Akhtar MF, Ma Q, Li Y, Chai W, Zhang Z, Li L, Wang C. Effect of Sperm Cryopreservation in Farm Animals Using Nanotechnology.. Animals (Basel) 2022 Sep 2;12(17).
- Najafi A, Mehdipour M, Mohammadi H, Mehdipour Z, Khorrami B, Nazari M. Effect of tempol and straw size on rooster sperm quality and fertility after post-thawing.. Sci Rep 2022 Jul 16;12(1):12192.
- Zhang R, Dong H, Zhao P, Shang C, Qi H, Ma Y, Gao C, Zhang D, Shen J, Lei Y, Jin Y, Lin P. Resveratrol and lycium barbarum polysaccharide improve Qinling giant panda (Ailuropoda melanoleuca Qinlingensis) sperm quality during cryopreservation.. BMC Vet Res 2022 Jan 7;18(1):23.
- Gimeno BF, Bariani MV, Laiz-Quiroga L, Martínez-León E, Von-Meyeren M, Rey O, Mutto AÁ, Osycka-Salut CE. Effects of In Vitro Interactions of Oviduct Epithelial Cells with Frozen-Thawed Stallion Spermatozoa on Their Motility, Viability and Capacitation Status.. Animals (Basel) 2021 Jan 3;11(1).
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