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Home / Equine Research Bank / J Androl / Reactive oxygen species and cryopreservation promote DNA fra...
Journal of andrology2003; 24(4); 621-628; doi: 10.1002/j.1939-4640.2003.tb02714.x

Reactive oxygen species and cryopreservation promote DNA fragmentation in equine spermatozoa.

Abstract: The objective of this study was to examine the effect of reactive oxygen species (ROS) and cryopreservation on DNA fragmentation of equine spermatozoa. In experiment 1, equine spermatozoa were incubated (1 hour, 38 degrees C) according to the following treatments: 1) sperm alone; 2) sperm + xanthine (X, 0.3 mM)-xanthine oxidase (XO, 0.025 U/mL); 3) sperm + X (0.6 mM)-XO (0.05 U/mL); and 4) sperm + X (1 mM)-XO (0.1 U/mL). In experiment 2, spermatozoa were incubated (1 hour, 38 degrees C) with X (1 mM)-XO (0.1 U/mL) and either catalase (200 U/mL), superoxide dismutase (SOD, 200 U/mL), or reduced glutathione (GSH, 10 mM). Following incubation, DNA fragmentation was determined by the single cell gel electrophoresis (comet) assay. In experiment 3, equine spermatozoa were cryopreserved, and DNA fragmentation was determined in fresh, processed, and postthaw sperm samples. In experiment 1, incubation of equine spermatozoa in the presence of ROS, generated by the X-XO system, increased DNA fragmentation (P <.005). In Experiment 2, the increase in DNA fragmentation associated with X-XO treatment was counteracted by the addition of catalase and GSH but not by SOD, suggesting that hydrogen peroxide and not superoxide appears to be the ROS responsible for such damage. In experiment 3, cryopreservation of equine spermatozoa was associated with an increase (P <.01) in DNA fragmentation when compared with fresh or processed samples. This study indicates that ROS and cryopreservation promote DNA fragmentation in equine spermatozoa; the involvement of ROS in cryopreservation-induced DNA damage remains to be determined.
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Publication Date: 2003-06-27 PubMed ID: 12826702DOI: 10.1002/j.1939-4640.2003.tb02714.xGoogle Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't
  • Research Support
  • U.S. Gov't
  • Non-P.H.S.

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 explores the effect of reactive oxygen species (ROS) and the process of cryopreservation on DNA fragmentation in horse sperm. The findings suggest that both ROS and cryopreservation can increase the potential for DNA damage, though the role of ROS in DNA damage during cryopreservation is yet to be identified.

Overview of Experiments and Procedures

  • The study consists mainly of three distinct experiments. Each experiment uses different combinations of chemicals including xanthine (X), xanthine oxidase (XO), catalase, superoxide dismutase (SOD), and glutathione (GSH) with horse sperm in order to assess the impact of these components on DNA fragmentation.
  • The first experiment studied four different conditions: sperm alone, sperm mixed with (X,0.3 mM)-XO (0.025 U/mL), sperm with (X,0.6 mM)-XO (0.05 U/mL) and sperm combined with (X,1 mM)-XO (0.1 U/mL). The aim was to understand the influence of ROS generated by the X-XO system on DNA fragmentation.
  • The second experiment involved the mixture of sperm with (X,1 mM)-XO (0.1 U/mL) and either catalase, superoxide dismutase (SOD), or reduced glutathione (GSH). The objective was to investigate if these substances could counteract the increase of DNA fragmentation associated with X-XO treatment.
  • The third experiment was intended to evaluate the impact of cryopreservation on DNA fragmentation in horse sperm.

Main Findings

  • From the first experiment, it was concluded that the presence of reactive oxygen species (ROS), generated by the X-XO system, triggered an increase in DNA fragmentation.
  • The second experiment revealed that the application of catalase and GSH could combat the impact of ROS on DNA fragmentation, whereas SOD seemed ineffective. This suggests that hydrogen peroxide, not superoxide, appears to be the ROS causing DNA damage.
  • The third experiment demonstrated that cryopreservation of horse sperm caused a significant increase in DNA fragmentation, when compared to fresh or processed samples.

Significance of the Study

  • This research helps expand the current understanding of the effects of ROS and cryopreservation on DNA fragmentation in equine spermatozoa.
  • The results imply that care should be taken when using ROS or cryopreservation in sperm preservation, due to the potential for DNA damage.
  • Further research will be needed to explore the role of ROS in cryopreservation-induced DNA damage.

Cite This Article

APA
Baumber J, Ball BA, Linfor JJ, Meyers SA. (2003). Reactive oxygen species and cryopreservation promote DNA fragmentation in equine spermatozoa. J Androl, 24(4), 621-628. https://doi.org/10.1002/j.1939-4640.2003.tb02714.x

Publication

Journal of andrology
ISSN: 0196-3635
NlmUniqueID: 8106453
Country: United States
Language: English
Volume: 24
Issue: 4
Pages: 621-628

Researcher Affiliations

Baumber, Julie
  • Departments of Population Health and Reproduction and Anatomy, Physiology, and Cell Biology, University of California, Davis, California 95616, USA.
Ball, Barry A
    Linfor, Jennifer J
      Meyers, Stuart A

        MeSH Terms

        • Animals
        • Antioxidants / pharmacology
        • Catalase / pharmacology
        • Cryopreservation
        • DNA Fragmentation
        • Glutathione / pharmacology
        • Horses
        • Male
        • Oxidative Stress / drug effects
        • Reactive Oxygen Species / metabolism
        • Semen Preservation / methods
        • Spermatozoa / cytology
        • Spermatozoa / metabolism
        • Superoxide Dismutase / pharmacology

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