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Home / Equine Research Bank / Am J Vet Res / Generation of reactive oxygen species by equine spermatozoa.
American journal of veterinary research2001; 62(4); 508-515; doi: 10.2460/ajvr.2001.62.508

Generation of reactive oxygen species by equine spermatozoa.

Abstract: To characterize generation of reactive oxygen species (ROS) by equine spermatozoa. Methods: Multiple semen samples collected from 9 stallions. Methods: Equine spermatozoa were separated from seminal plasma on a discontinuous polyvinylpyrrolidone (PVP)-coated silica gradient and resuspended in a modified Tyrode albumin-lactate-pyruvate medium. Amount of hydrogen peroxide (H2O2) generated was assayed by use of a 1-step fluorometric assay, using 10-acetyl-3,7-dihydroxyphenoxazine as a probe for detection of H2O2 in a microplate assay format. Concentration of H2O2 was determined by use of a fluorescence microplate reader. Results: Amount of H2O2 generated increased significantly with time and spermatozoa concentration for live and flash-frozen spermatozoa, and amount of H2O2 generated was significantly greater for flash-frozen than for live spermatozoa. Addition of the reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) significantly increased generation of H2O2 by live and flash-frozen spermatozoa. Addition of a calcium ionophore also significantly increased the amount of H2O2 generated by live spermatozoa but did not have an effect on amount of H2O2 generated by flash-frozen spermatozoa. Abnormal equine spermatozoa generated significantly greater amounts of H2O2 than did normal spermatozoa. Conclusions: Equine spermatozoa generate ROS in vitro, possibly via a NADPH-oxidase reaction. Spermatozoa damaged during flash-freezing or morphologically abnormal spermatozoa generated significantly greater amounts of ROS than did live or morphologically normal spermatozoa. Damaged and abnormal spermatozoa generate greater amounts of ROS that may contribute to reduced fertility or problems related to semen preservation.
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Publication Date: 2001-05-01 PubMed ID: 11327456DOI: 10.2460/ajvr.2001.62.508Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • 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.

This research study explores the creation of reactive oxygen species (ROS) by horse sperm cells and how this process might be affected by factors such as cell morphology, cellular damage, and artificial preservation techniques.

Methodology

  • The study involved multiple semen samples collected from nine horses. The sperm cells were separated from the seminal fluid using a polyvinylpyrrolidone-coated silica gradient and then submerged in a special medium.
  • To measure the amount of hydrogen peroxide (H2O2), a type of ROS, produced by the sperm cells, a fluorometric assay was employed. This method utilizes a specific probe, known as 10-acetyl-3,7-dihydroxyphenoxazine, that can detect H2O2 in a microplate format. The concentration of H2O2 was then determined using a fluorescence microplate reader.

Findings

  • The study found that the production of H2O2 rose significantly over time and was positively correlated with the concentration of the sperm cells. This was true for both live and frozen sperm cells, with the latter producing higher amounts of H2O2.
  • Adding a reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) notably boosted the production of H2O2 in both live and frozen sperm cells. Introducing a calcium ionophore also increased H2O2 production in live sperm cells, but did not have the same effect on frozen ones.
  • The research revealed that abnormal sperm cells produced significantly more H2O2 than normal ones.

Conclusions

  • The researchers concluded that horse spermatozoa generate ROS, possibly through a reaction involving NADPH oxidase. This process was found to be more pronounced in sperm cells that were either damaged during freezing or morphologically abnormal.
  • The overproduction of ROS could potentially compromise fertility or create difficulties in sperm preservation. Therefore, understanding the mechanisms behind ROS production and finding ways to control it could have important implications for breeding practices and fertility treatments in horses.

Cite This Article

APA
Ball BA, Vo AT, Baumber J. (2001). Generation of reactive oxygen species by equine spermatozoa. Am J Vet Res, 62(4), 508-515. https://doi.org/10.2460/ajvr.2001.62.508

Publication

American journal of veterinary research
ISSN: 0002-9645
NlmUniqueID: 0375011
Country: United States
Language: English
Volume: 62
Issue: 4
Pages: 508-515

Researcher Affiliations

Ball, B A
  • Department of Population Health and Reproduction, School of Veterinary Medicine, University of California, Davis 95616, USA.
Vo, A T
    Baumber, J

      MeSH Terms

      • Animals
      • Calcimycin / pharmacology
      • Catalase / pharmacology
      • Centrifugation, Density Gradient / veterinary
      • Cryopreservation / veterinary
      • Horses / metabolism
      • Hydrogen Peroxide / analysis
      • Hydrogen Peroxide / antagonists & inhibitors
      • Hydrogen Peroxide / metabolism
      • Ionophores / pharmacology
      • Male
      • Microscopy, Interference / veterinary
      • N-Formylmethionine Leucyl-Phenylalanine / pharmacology
      • NADP / pharmacology
      • Reactive Oxygen Species / metabolism
      • Semen Preservation / adverse effects
      • Semen Preservation / veterinary
      • Spermatozoa / drug effects
      • Spermatozoa / metabolism
      • Superoxides / metabolism
      • Tetradecanoylphorbol Acetate / pharmacology

      Citations

      This article has been cited 25 times.
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