Determination of glutathione peroxidase and superoxide dismutase-like activities in equine spermatozoa, seminal plasma, and reproductive tissues.
Abstract: To determine glutathione peroxidase (GPX) and superoxide dismutase (SOD)-like activities in spermatozoa, seminal plasma, and reproductive tissues (ie, testis, epididymis, bulbourethral gland, prostate, vesicular gland, and ampulla) in horses. Methods: Seminal plasma from 17 stallions, spermatozoa from 5 stallions, and reproductive tissues from 3 stallions. Methods: Activity of GPX was determined by use of assays measuring oxidation of NADPH in the presence of exogenous glutathione, cumene hydroperoxide, and glutathione reductase. Activity of SOD-like enzymes was determined by use of the nitroblue tetrazolium assay. Results: Mean GPX and SOD-like activities in seminal plasma were 1.3 +/- 0.1 nmol of NADPH oxidized/min/mg of protein and 29.2 +/- 6.6 U/mg of protein, respectively. Mean GPX activities in spermatozoa separated from seminal plasma by centrifugation and via Percoll gradient were 2.2 +/- 0.3 nmol and 6.1 +/- 1.3 nmol of NADPH oxidized/min/mg of protein, respectively. Mean SOD-like activity of spermatozoa separated by centrifugation was 58.6 +/- 22.3 U/mg of protein; SOD-like activity was not detected in Percoll-separated spermatozoa. Among reproductive tissues, the ampulla and prostate had the highest SOD-like activity, although this was not significantly different from activity in other tissues. Testes and spermatozoa from the cauda epididymis contained significantly more GPX activity than other tissues. Conclusions: Results suggest that although equine seminal plasma contains high SOD-like enzyme activity, spermatozoa have limited GPX and SOD-like activity. Enzymatic antioxidant activity in equine spermatozoa appears to be predominantly derived from seminal plasma adsorbed onto the plasma membrane. Removal of seminal plasma during semen processing may increase oxidative stress in equine-spermatozoa.
Publication Date: 2005-09-22 PubMed ID: 16173486DOI: 10.2460/ajvr.2005.66.1415Google 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
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The research aimed at measuring and comparing the activities of glutathione peroxidase (GPX) and superoxide dismutase (SOD)-like enzymes in horse sperm, seminal fluid, and reproductive tissues. It found that these enzymes’ activity, essential to handling oxidative stress, varies across the tested biological samples, with seminal plasma showing high SOD-like activity while sperm shows limited GPX and SOD-like activity.
Methodology
- The researchers collected seminal plasma from 17 stallions and sperm from 5 stallions. They also examined reproductive tissues (testis, epididymis, bulbourethral gland, prostate, vesicular gland, and ampulla) from 3 stallions.
- To measure the activity of GPX, they employed assays that calculate the oxidation of NADPH in the presence of exogenous glutathione, cumene hydroperoxide, and glutathione reductase.
- SOD-like enzyme activity was determined using the nitroblue tetrazolium assay.
Results
- The mean GPX and SOD-like activities in seminal plasma were found to be 1.3 ± 0.1 nmol of NADPH oxidized/min/mg of protein and 29.2 ± 6.6 U/mg of protein, respectively.
- The average GPX activities in sperm separated from seminal plasma through centrifugation and Percoll gradient were found to be 2.2 ± 0.3 nmol and 6.1 ± 1.3 nmol of NADPH oxidized/min/mg of protein, respectively.
- The mean SOD-like activity in sperm separated by centrifugation was 58.6 ± 22.3 U/mg of protein. However, SOD-like activity could not be detected in Percoll-separated sperm.
- Of all the reproductive tissues tested, the ampulla and prostate tissues showed the highest SOD-like activity, while testes and sperm from the cauda epididymis had significantly higher GPX activity.
Conclusion
- From the research, it is suggested that equine seminal plasma contains a high amount of SOD-like enzyme activity. In contrast, sperm shows limited activity of GPX and SOD-like enzymes.
- The enzymatic antioxidant activity in equine sperm seems to be mainly derived from seminal plasma adsorbed onto the plasma membrane. As a result, removal of seminal plasma during semen processing may subsequently increase oxidative stress in horse sperm.
Cite This Article
APA
Baumber J, Ball BA.
(2005).
Determination of glutathione peroxidase and superoxide dismutase-like activities in equine spermatozoa, seminal plasma, and reproductive tissues.
Am J Vet Res, 66(8), 1415-1419.
https://doi.org/10.2460/ajvr.2005.66.1415 Publication
Researcher Affiliations
- Department of Population Health and Reproduction, School of Veterinary Medicine, University of California, Davis, CA 95616, USA.
MeSH Terms
- Animals
- Bulbourethral Glands / enzymology
- Genitalia, Male / enzymology
- Glutathione Peroxidase / metabolism
- Horses / metabolism
- Male
- Prostate / enzymology
- Semen / enzymology
- Spermatozoa / enzymology
- Superoxide Dismutase / metabolism
- Testis / enzymology
Citations
This article has been cited 12 times.- Ayad B, Omolaoye TS, Louw N, Ramsunder Y, Skosana BT, Oyeipo PI, Du Plessis SS. Oxidative Stress and Male Infertility: Evidence From a Research Perspective.. Front Reprod Health 2022;4:822257.
- Catalán J, Yánez-Ortiz I, Tvarijonaviciute A, González-Aróstegui LG, Rubio CP, Barranco I, Yeste M, Miró J. Seminal Plasma Antioxidants Are Related to Sperm Cryotolerance in the Horse.. Antioxidants (Basel) 2022 Jun 28;11(7).
- Palacin-Martinez C, Alvarez M, Montes-Garrido R, Neila-Montero M, Anel-Lopez L, de Paz P, Anel L, Riesco MF. Frequency of Semen Collection Affects Ram Sperm Cryoresistance.. Animals (Basel) 2022 Jun 8;12(12).
- Catalán J, Yánez-Ortiz I, Tvarijonaviciute A, González-Arostegui LG, Rubio CP, Yeste M, Miró J, Barranco I. Impact of Seminal Plasma Antioxidants on Donkey Sperm Cryotolerance.. Antioxidants (Basel) 2022 Feb 18;11(2).
- Miró J, Catalán J, Marín H, Yánez-Ortiz I, Yeste M. Specific Seminal Plasma Fractions Are Responsible for the Modulation of Sperm-PMN Binding in the Donkey.. Animals (Basel) 2021 May 13;11(5).
- Khaki A, Araghi A, Lotfi M, Nourian A. Differences between some biochemical components in seminal plasma of first and second ejaculations in dual-purpose Simmental (Fleckvieh) bulls and their relationships with semen quality parameters.. Vet Res Forum 2021 Winter;12(1):39-46.
- Peña FJ, O'Flaherty C, Ortiz Rodríguez JM, Martín Cano FE, Gaitskell-Phillips GL, Gil MC, Ortega Ferrusola C. Redox Regulation and Oxidative Stress: The Particular Case of the Stallion Spermatozoa.. Antioxidants (Basel) 2019 Nov 19;8(11).
- Papas M, Catalán J, Fernandez-Fuertes B, Arroyo L, Bassols A, Miró J, Yeste M. Specific Activity of Superoxide Dismutase in Stallion Seminal Plasma Is Related to Sperm Cryotolerance.. Antioxidants (Basel) 2019 Nov 9;8(11).
- Heidari Nasirabadi M, Shirazi A, Kadivar A, Shams-Esfandabadi N, Mohebbi A, Ahmadi E. Sericin Ameliorates the Capacitation State and Chromatin Integrity of Frozen-Thawed Stallion Spermatozoa by Reducing Oxidative Stress.. Avicenna J Med Biotechnol 2019 Jul-Sep;11(3):245-252.
- Falchi L, Galleri G, Dore GM, Zedda MT, Pau S, Bogliolo L, Ariu F, Pinna A, Nieddu S, Innocenzi P, Ledda S. Effect of exposure to CeO(2) nanoparticles on ram spermatozoa during storage at 4 °C for 96 hours.. Reprod Biol Endocrinol 2018 Mar 6;16(1):19.
- Dzyuba V, Cosson J, Dzyuba B, Yamaner G, Rodina M, Linhart O. The antioxidant system of seminal fluid during in vitro storage of sterlet Acipenser ruthenus sperm.. Fish Physiol Biochem 2016 Apr;42(2):563-8.
- Dzyuba V, Dzyuba B, Cosson J, Boryshpolets S, Yamaner G, Kholodniy V, Rodina M. The antioxidant system of sterlet seminal fluid in testes and Wolffian ducts.. Fish Physiol Biochem 2014 Dec;40(6):1731-9.
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