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Home / Equine Research Bank / Theriogenology / Reactive oxygen species promote tyrosine phosphorylation and...
Theriogenology2003; 60(7); 1239-1247; doi: 10.1016/s0093-691x(03)00144-4

Reactive oxygen species promote tyrosine phosphorylation and capacitation in equine spermatozoa.

Abstract: The objective of this study was to examine the influence of reactive oxygen species (ROS) on equine sperm capacitation. Motile equine spermatozoa were separated on a discontinuous Percoll gradient, resuspended at 10 x 10(6)ml in Tyrode's medium supplemented with BSA (0.5%) and polyvinyl alcohol (0.5%) and incubated at 39 degrees C for 2h with or without the xanthine (X; 0.1mM)-xanthine oxidase (XO; 0.01 U/ml) system or NADPH (0.25 mM). The importance of hydrogen peroxide or superoxide for capacitation was determined by the addition of catalase (CAT; 150 U/ml) or superoxide dismutase (SOD; 150 U/ml), respectively. Following incubation, acrosomal exocytosis was induced by a 5 min incubation at 39 degrees C with progesterone (3.18 microM), and sperm viability and acrosomal integrity were then determined by staining with Hoechst 33258 and fluoroisothiocyanate-conjugated Pisum sativum agglutin. To examine tyrosine phosphorylation, treatments were subjected to sodium dodecyl sulfate-polyacrylaminde gel electrophoresis (SDS-PAGE) followed by Western blot analysis with the anti-phosphotyrosine antibody (alpha-PY; clone 4G10). Capacitation with the X-XO system or NADPH led to a significant (P<0.0001) increase in live acrosome-reacted spermatozoa compared to controls. The addition of CAT or SOD prevented the increase in live acrosome-reacted spermatozoa associated with X-XO treatment. Incubation with the X-XO system was also associated with a significant (P<0.005) increase in tyrosine phosphorylation when compared to controls, which could be prevented by the addition of CAT but not SOD. This study indicates that ROS can promote equine sperm capacitation and tyrosine phosphorylation, suggesting a physiological role for ROS generation by equine spermatozoa.
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Publication Date: 2003-09-27 PubMed ID: 14511778DOI: 10.1016/s0093-691x(03)00144-4Google 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 investigates the effect of reactive oxygen species (ROS) on the capacitation and tyrosine phosphorylation of equine sperm cells. The findings suggest that ROS assist in these processes, indicating a potential physiological role for ROS production in equine sperm.

Research Methodology

  • The study involved examination of motile equine spermatozoa. These live sperm cells were segregated on a Percoll gradient, a concoction designed to separate particles based on their size and density.
  • The separated sperm cells were suspended in Tyrode’s medium, a solution designed to maintain the ion balance in cells and supplemented with Bovine Serum Albumin (BSA) and polyvinyl alcohol. This was meant to mimic the natural environment of the sperm cells.
  • These cells were then subjected to two-hour incubation at 39 degrees Celcius, both with and without the xanthine/xanthine oxidase (X/XO) system or NADPH, chemicals known to induce ROS.
  • The researchers used catalase and superoxide dismutase (enzymes that neutralize hydrogen peroxide and superoxide respectively) to determine the importance of these ROS in capacitation.
  • Subsequent to incubation, acrosomal exocytosis (the process by which the sperm cell prepares itself for fusion with an egg cell) was stimulated using progesterone, a steroid hormone.
  • The survival rate of the sperm cells and their acrosomal integrity (the structural soundness of the sperm cell’s head where genetic material is stored) were determined using a stain called Hoechst 33258 and fluoroisothiocyanate-conjugated Pisum sativum agglutin.

Key Findings

  • Compared to the control group, using the X-XO system or NADPH to induce capacitation led to a significant increase in live acrosome-reacted (acrosome exocytosis had occurred and remained viable) spermatozoa.
  • This increase in live acrosome-reacted spermatozoa could be prevented by adding catalase or superoxide dismutase, indicating the importance of hydrogen peroxide and superoxide, types of ROS, in the capacitation process.
  • Incubation with the X-XO system resulted in a significant increase in tyrosine phosphorylation (a process that potentially makes the sperm cells more active) when compared to the control samples. This could also be prevented by the addition of catalase, but not superoxide dismutase.
  • The important conclusion of this research is that ROS seems to play a significant role in promoting equine sperm capacitation and tyrosine phosphorylation. This suggests that equine spermatozoa may physiologically generate ROS for these reproductive processes.

Cite This Article

APA
Baumber J, Sabeur K, Vo A, Ball BA. (2003). Reactive oxygen species promote tyrosine phosphorylation and capacitation in equine spermatozoa. Theriogenology, 60(7), 1239-1247. https://doi.org/10.1016/s0093-691x(03)00144-4

Publication

Theriogenology
ISSN: 0093-691X
NlmUniqueID: 0421510
Country: United States
Language: English
Volume: 60
Issue: 7
Pages: 1239-1247

Researcher Affiliations

Baumber, J
  • Department of Population Health & Reproduction, 1114 Tupper Hall, University of California, Davis, CA 95616, USA.
Sabeur, K
    Vo, A
      Ball, B A

        MeSH Terms

        • Animals
        • Blotting, Western
        • Catalase / administration & dosage
        • Catalase / metabolism
        • Electrophoresis, Polyacrylamide Gel
        • Horses
        • Hydrogen Peroxide / metabolism
        • Hydrogen Peroxide / pharmacology
        • Male
        • NADP / administration & dosage
        • Phosphorylation
        • Reactive Oxygen Species / pharmacology
        • Sperm Capacitation / drug effects
        • Spermatozoa / drug effects
        • Spermatozoa / metabolism
        • Superoxide Dismutase / administration & dosage
        • Superoxide Dismutase / metabolism
        • Superoxides / metabolism
        • Superoxides / pharmacology
        • Tyrosine / metabolism
        • Xanthine / administration & dosage
        • Xanthine Oxidase / administration & dosage

        Citations

        This article has been cited 15 times.
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