Irradiating frozen-thawed stallion sperm with red-light increases their resilience to withstand post-thaw incubation at 38 °C.
Abstract: The aim of this study was to evaluate whether red-light stimulation increases the longevity and resilience of cryopreserved stallion sperm to withstand post-thaw incubation for 120 min. Sixteen frozen straws of 0.5 mL from eight stallions were used. Samples were cryopreserved, thawed through incubation at 38 °C for 30 s and divided into the control and samples exposed to red-light using a triple LED photo-activation system (wavelength: 620-630 nm). Three irradiation protocols consisting of different light-dark-light intervals (1-1-1, 2-2-2 and 3-3-3 min) were tested. Sperm quality parameters were analyzed immediately after light-stimulation (0 min) and after 120 min of incubation at 38 °C. Sperm motility was evaluated using a Computerized Semen Analysis System (CASA), and flow cytometry and different fluorochromes were used to evaluate the integrity and lipid disorder of plasma membrane, mitochondrial membrane potential and intracellular levels of peroxides and superoxides. Irradiation significantly increased the percentages of spermatozoa with high mitochondrial membrane potential (1-1-1 pattern) and the intracellular levels of peroxides (2-2-2 pattern) at 0 min. In addition, sperm kinematic parameters (2-2-2 and 3-3-3 patterns) and percentages of viable spermatozoa with low membrane lipid disorder (3-3-3 pattern) were significantly higher in irradiated samples than in the control at 120 min. Our results indicate that red-light stimulation could help increase the resilience of frozen-thawed stallion sperm to withstand post-thaw incubation at 38 °C for 120 min and that these effects rely on the irradiation pattern. Further research should evaluate whether light-stimulation could also have a positive on fertility rates after artificial insemination.
Copyright © 2020 Elsevier Inc. All rights reserved.
Publication Date: 2020-07-28 PubMed ID: 32805646DOI: 10.1016/j.theriogenology.2020.07.027Google Scholar: Lookup
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- Journal Article
Summary
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This research investigates if exposing cryopreserved (frozen, then thawed) stallion sperm to red light can heighten their survivability, particularly by enhancing their ability to endure a 120-minute post-thaw incubation at 38 degrees Celsius.
Study Methodology
- The research utilized frozen sperm samples from eight stallions, with a total of 16 0.5 mL straws.
- The samples underwent cryopreservation, were thawed via incubation at 38 degrees Celsius for 30 seconds, then segregated into a control group and red-light exposed samples.
- The research employed a triple LED photo-activation system emitting red light, with a wavelength of 620-630 nm.
- The irradiation protocols consisted of three distinct light-dark-light intervals lasting 1-1-1, 2-2-2, and 3-3-3 minutes, respectively.
- Sperm quality parameters were analyzed immediately after light-stimulation and after incubation at 38 degrees Celsius for 120 minutes.
Sperm Evaluation Techniques
- Sperm motility was assessed using a Computerized Semen Analysis System (CASA).
- Flow cytometry and different fluorochromes were used to evaluate the integrity and lipid disorder of the plasma membrane, mitochondrial membrane potential, and intracellular levels of peroxides and superoxides.
Research Findings
- Significant increases were observed in the percentages of spermatozoa with high mitochondrial membrane potential (1-1-1 pattern) and intracellular levels of peroxides (2-2-2 pattern) at 0 minutes post-irradiation.
- After 120 minutes, sperm kinematic parameters (2-2-2 and 3-3-3 patterns), and the percentages of viable spermatozoa with low membrane lipid disorder (3-3-3 pattern) were proven to be significantly higher in irradiated samples than in the control group.
- Suggesting that red-light stimulation could potentially enhance the survivability of frozen-thawed stallion sperm post-thaw incubation at 38 degrees Celsius for 120 minutes, with effects varying based on the irradiation pattern used.
Future Research
- As an extension of this study, the researchers propose exploring whether light-stimulation could also positively impact fertility rates after artificial insemination.
Cite This Article
APA
Catalán J, Llavanera M, Bonilla-Correal S, Papas M, Gacem S, Rodríguez-Gil JE, Yeste M, Miró J.
(2020).
Irradiating frozen-thawed stallion sperm with red-light increases their resilience to withstand post-thaw incubation at 38 °C.
Theriogenology, 157, 85-95.
https://doi.org/10.1016/j.theriogenology.2020.07.027 Publication
Researcher Affiliations
- Unit of Animal Reproduction, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Autonomous University of Barcelona, Spain; Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, Girona, Spain; Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, Girona, Spain.
- Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, Girona, Spain; Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, Girona, Spain.
- Unit of Animal Reproduction, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Autonomous University of Barcelona, Spain; Faculty of Veterinary Medicine, Antonio Nariño University, Bogotá, Colombia.
- Unit of Animal Reproduction, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Autonomous University of Barcelona, Spain.
- Unit of Animal Reproduction, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Autonomous University of Barcelona, Spain.
- Unit of Animal Reproduction, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Autonomous University of Barcelona, Spain.
- Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, Girona, Spain; Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, Girona, Spain. Electronic address: marc.yeste@udg.edu.
- Unit of Animal Reproduction, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Autonomous University of Barcelona, Spain.
MeSH Terms
- Animals
- Cryopreservation / veterinary
- Freezing
- Horses
- Male
- Semen Preservation / veterinary
- Sperm Motility
- Spermatozoa
Conflict of Interest Statement
Declaration of competing interest J.E.R.G. and M.Y. are inventors of a patent entitled ‘Method and apparatus for improving the quality of mammalian sperm’ (European Patent Office, No. 16199093.2; EP-3-323-289-A1), which is owned by Instruments Utils de Laboratori Geniul, SL (Barcelona, Spain).
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