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Reactive oxygen species promote tyrosine phosphorylation and capacitation in equine spermatozoa.
Theriogenology 2003; 60(7):1239-47T

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.

Authors+Show Affiliations

Department of Population Health & Reproduction, 1114 Tupper Hall, University of California, Davis, CA 95616, USA.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

14511778

Citation

Baumber, J, et al. "Reactive Oxygen Species Promote Tyrosine Phosphorylation and Capacitation in Equine Spermatozoa." Theriogenology, vol. 60, no. 7, 2003, pp. 1239-47.
Baumber J, Sabeur K, Vo A, et al. Reactive oxygen species promote tyrosine phosphorylation and capacitation in equine spermatozoa. Theriogenology. 2003;60(7):1239-47.
Baumber, J., Sabeur, K., Vo, A., & Ball, B. A. (2003). Reactive oxygen species promote tyrosine phosphorylation and capacitation in equine spermatozoa. Theriogenology, 60(7), pp. 1239-47.
Baumber J, et al. Reactive Oxygen Species Promote Tyrosine Phosphorylation and Capacitation in Equine Spermatozoa. Theriogenology. 2003 Oct 15;60(7):1239-47. PubMed PMID: 14511778.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Reactive oxygen species promote tyrosine phosphorylation and capacitation in equine spermatozoa. AU - Baumber,J, AU - Sabeur,K, AU - Vo,A, AU - Ball,B A, PY - 2003/9/27/pubmed PY - 2003/12/3/medline PY - 2003/9/27/entrez SP - 1239 EP - 47 JF - Theriogenology JO - Theriogenology VL - 60 IS - 7 N2 - 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. SN - 0093-691X UR - https://www.unboundmedicine.com/medline/citation/14511778/Reactive_oxygen_species_promote_tyrosine_phosphorylation_and_capacitation_in_equine_spermatozoa_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0093691X03001444 DB - PRIME DP - Unbound Medicine ER -