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Home / Equine Research Bank / J Androl / The effect of reactive oxygen species on equine sperm motili...
Journal of andrology2000; 21(6); 895-902;

The effect of reactive oxygen species on equine sperm motility, viability, acrosomal integrity, mitochondrial membrane potential, and membrane lipid peroxidation.

Abstract: The objective of this study was to examine the influence of reactive oxygen species (ROS), generated through the use of the xanthine (X)-xanthine oxidase (XO) system, on equine sperm motility, viability, acrosomal integrity, mitochondrial membrane potential, and membrane lipid peroxidation. Equine spermatozoa were separated from seminal plasma on a discontinuous Percoll gradient, and spermatozoa were incubated with 0.6 mM X and 0.05 U/mL XO for 30 minutes. Catalase (150 U/mL), superoxide dismutase (SOD, 150 U/mL), or glutathione (GSH, 1.5 mM) were evaluated for their ability to preserve sperm function in the presence of the induced oxidative stress. At the end of the 30-minute incubation, sperm motility was determined by computer-assisted semen analysis. Viability and acrosomal integrity were determined by Hoechst-Pisum sativum staining, and mitochondrial membrane potential was determined by staining with JC-1. Incubation with the X-XO system led to a significant (P < .01) increase in hydrogen peroxide production and an associated decrease (P < .01) in motility parameters. Total motility was significantly (P < .01) lower in the presence of X-XO compared with the case of the control (29%+/-9% vs 73%+/-1%, respectively). Catalase, but not SOD, prevented a decline in motility secondary to oxidative stress (71%+/-4% vs 30%+/-3%, respectively). The addition of glutathione had an intermediate effect in preserving sperm motility at the end of the 30-minute incubation (53%+/-3%). No influence of X-XO could be determined on viability, acrosomal integrity, or mitochondrial membrane potential. In order to promote lipid peroxidation, samples were incubated with ferrous sulfate (0.64 mM) and sodium ascorbate (20 mM) for 2 hours after the X-XO incubation. No increase in membrane lipid peroxidation was detected. This study indicates that hydrogen peroxide is the major ROS responsible for damage to equine spermatozoa. The decrease in sperm motility associated with ROS occurs in the absence of any detectable decrease in viability, acrosomal integrity, or mitochondrial membrane potential or of any detectable increase in lipid peroxidation.
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Publication Date: 2000-12-06 PubMed ID: 11105916
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  • 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 study investigates the effect of reactive oxygen species (ROS) on equine sperm processes like motility, viability, and acrosomal integrity. The study concludes that hydrogen peroxide, a type of ROS, significantly hinders sperm motility without noticeably affecting other sperm attributes such as viability, acrosomal integrity, or mitochondrial membrane potential.

Study Methodology

  • The researchers separated equine spermatozoa from seminal plasma on a discontinuous Percoll gradient. They then incubated the spermatozoa with xanthine (X) and xanthine oxidase (XO), intending to generate ROS.
  • Next, they evaluated the ability of catalase, superoxide dismutase (SOD), and glutathione (GSH) to preserve sperm function in the presence of the induced oxidative stress.
  • The research team used computer-assisted semen analysis to determine sperm motility after the 30-minute incubation. Viability and acrosomal integrity were assessed by employing Hoechst-Pisum sativum staining, and mitochondrial membrane potential was determined by staining with JC-1.

Study Findings

  • The incubation with X-XO led to a significant increase in hydrogen peroxide production and a corresponding decrease in motility parameters.
  • Total motility was significantly lower with X-XO than in control groups.
  • Catalase prevented declined motility due to oxidative stress, while SOD did not demonstrate a similar effect. Glutathione showed a moderate ability to preserve sperm motility.
  • The X-XO system had no significant effect on viability, acrosomal integrity, or mitochondrial membrane potential.
  • The research team attempted to promote lipid peroxidation by incubating the samples with ferrous sulfate and sodium ascorbate post X-XO incubation, however, no increase in membrane lipid peroxidation was witnessed.

Study Conclusion

This research indicates that hydrogen peroxide, a type of ROS, is primarily responsible for damage to equine spermatozoa. Particularly, it has the significant effect of decreasing sperm motility. The study didn’t find any noticeable effect on sperm viability, acrosomal integrity, mitochondrial membrane potential, or lipid peroxidation. This suggests that the detrimental effects of ROS on equine sperm are relatively specific, primarily impacting sperm motility.

Cite This Article

APA
Baumber J, Ball BA, Gravance CG, Medina V, Davies-Morel MC. (2000). The effect of reactive oxygen species on equine sperm motility, viability, acrosomal integrity, mitochondrial membrane potential, and membrane lipid peroxidation. J Androl, 21(6), 895-902.

Publication

Journal of andrology
ISSN: 0196-3635
NlmUniqueID: 8106453
Country: United States
Language: English
Volume: 21
Issue: 6
Pages: 895-902

Researcher Affiliations

Baumber, J
  • Department of Population Health and Reproduction, University of California, Davis 95616, USA.
Ball, B A
    Gravance, C G
      Medina, V
        Davies-Morel, M C

          MeSH Terms

          • Acrosome / physiology
          • Acrosome / ultrastructure
          • Animals
          • Catalase / metabolism
          • Cell Survival
          • Glutathione / metabolism
          • Horses
          • In Vitro Techniques
          • Intracellular Membranes / physiology
          • Male
          • Membrane Potentials
          • Mitochondria / physiology
          • Reactive Oxygen Species
          • Sperm Motility
          • Spermatozoa / cytology
          • Spermatozoa / drug effects
          • Spermatozoa / physiology
          • Superoxide Dismutase / metabolism
          • Xanthine / metabolism
          • Xanthine / pharmacology
          • Xanthine Oxidase / metabolism
          • Xanthine Oxidase / pharmacology

          Citations

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