Improvement of Ram’s Frozen Sperm Quality Using Glutathione Antioxidant

Document Type : Full Research Paper

Authors

1 Animal Science Research Institute of Iran (ASRI), Agricultural Research Education and Extension Organization (AREEO), Karaj, Iran

2 Department of Animal Science, University College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran

Abstract

The aim of this study was improvement of ram frozen semen efficiency using addition of glutathione antioxidant to freezing medium. In this experiment, semen samples were collected from five Zell rams and add to the extender containing 0, 0.5, 1, 2, 4 and 8 mM glutathione. In sperm quality evaluation, sperm samples were evaluated and motion parameters, membrane integrity, morphology, acrosome integrity, mitochondria activity, viability and lipid peroxidation were assessed. The results showed using 4 mM glutathione resulted in improving of sperm total and progressive motility, membrane integrity, mitochondria activity, acrosome integrity, and viability as well as reduction of apoptosis rate and lipid peroxidation. Using the treatment of 8 mM glutathione caused to reduction of sperm quality after thawing and the antioxidantd did not have any effect on sperm morphology. In conclusion, using 4 mM glutathione in freezing extender could be a suitable method to improve ram sperm reproductive performance after freeze-thaw process.

Keywords


1 - Ansari, M.S., B.A. Rakha, N. Ullah, S.M.H. Andrabi and et al. 2010. Effect of exogenous glutathione in extender on the freezability of Nili-Ravi buffalo (Bubalus bubalis) bull spermatozoa. Animal Science Reports. 28: 235–244.
2 - Armstrong, J.S., M. Rajasekaran, W. Chamulitrat, P. Gatti and et al. 1999. Characterization of reactive oxygen species induced effects on human spermatozoa movement and energy metabolism. Free Radical Biology and Medicine. 26: 869–880.
3 - Bailey, J.L., J.F. Bilodeau and N. Cormier. 2000. Semen cryopreservation in domestic animals: a damaging and capacitating phenomenon. Journal of Andrology. 21: 1–7.
4 - Bartoov, B., F. Eltes, R. Weissenberg and B. Lunenfeld. 1980. Morphological characterization of abnormal human spermatozoa using transmission electron microscopy. Archives of Andrology. 5: 305–322.
5 - Bilodeau, J.F., S. Blanchette, C. Gagnon and M.A. Sirard. 2001. Thiols Prevent H2O2-mediated loss of sperm motility in cryopreserved bull semen. Theriogenology. 56: 275–286.
6 - Bucak, M.N. and N. Tekin. 2007. Protective effect of taurine, glutathione and trehalose on the liquid storage of ram semen. Small Ruminant Research. 73: 103–108.
7 - Chenoweth P.J. 2005. Genetic sperm defects. Theriogenology. 457–468.
8 - Estrada, E., J.E. Rodríguez-Gil, L.G. Rocha, S. Balasch and et al. 2014. Supplementing cryopreservation media with reduced glutathione increases fertility and prolificacy of sows inseminated with frozen-thawed boar semen. Andrology. 2: 88–99.
9 - Fattah, A., M. Sharafi, R. Masoudi, A. Shahverdi and et al. 2017. L-Carnitine in rooster semen cryopreservation: Flow cytometric, biochemical and motion findings for frozen-thawed sperm. Cryobiology. 74: 148–153.
10 - Gadea, J., D. Gumbao, C. Matás and R. Romar. 2005. Supplementation of the thawing media with reduced glutathione improves function and the in vitro fertilizing ability of boar spermatozoa after cryopreservation, Journal of Andrology. 26: 749–756.
11 - Gadea, J., M. Molla, E. Selles, M.A. Marco and et al. 2011. Reduced glutathione content in human sperm is decreased after cryopreservation: Effect of the addition of reduced glutathione to the freezing and thawing extenders. Cryobiology. 62: 40–46.
12 - Gravance, C.G., D.L. Garner, M.G. Miller and T. Berger. 2000. Fluorescent probes and flow cytometry to assess rat sperm integrity and mitochondrial function. Reproductive Toxicology. 15: 5–10.
13 - Lotfi, S., M. Mehri, M. Sharafi and R. Masoudi. 2017. Hyaluronic acid improves frozen-thawed sperm quality and fertility potential in rooster. Animal Reproduction Science. 184: 204–210.
14 - Lu, S.C. 2013. Glutathione synthesis. Biochimica et Biophysica Acta (BBA)-General Subjects. 1830: 3143–53.
15 - Masoudi, R., M. Sharafi, A. Zareh Shahneh, A. Towhidi and et al. 2016. Fertility and flow cytometry study of frozen-thawed sperm in cryopreservation medium supplemented with soybean lecithin. Cryobiology. 73: 69–72.
16 - Masoudi, R., M. Sharafi, A. Zareh Shahneh, A. Towhidi and et al. 2016. Effect of dietary fish oil supplementation on ram semen freeze ability and fertility using soybean lecithin– and egg yolk–based extenders. Theriogenology. 86: 1583–1588.
17 -Masoudi, R., A. Zare Shahneh, A. Towhidi and et al. 2017. Fertility response of artificial insemination methods in sheep with fresh and frozen-thawed semen Cryobiology. 74: 77–80.
18 - Meseguer, M., N. Garrido, J.A. Martínez-Conejero, C. Simón and et al. 2004. Role of cholesterol, calcium, and mitochondrial activity in the susceptibility for cryodamage after a cycle of freezing and thawing, Fertility and Sterility. 81: 588–594.
19 -Revell, S.G. and R.A. Mrode. 1994. An osmotic resistance test for bovine semen. Animal Reproduction Science. 36: 77–86.
20 - Salmani, H., M.M. Nabi, H. Vaseghi-Dodaran, M.B. Rahman and et al. 2013. Effect of glutathione in soybean lecithin-based semen extender on goat semen quality after freeze-thawing. Small Ruminant Research. 112: 123–127.
21 - Sharafi, M., M. Forouzanfar, S.M. Hosseini, M. Hajian and et al. 2009. In vitro comparison of soybean lecithin based-extender with commercially available extender for ram semen cryopreservation. International Journal of Fertility and Sterility. 3: 149–152.
22 - Lector, C. 1996. Oxidative stress and role of antioxidants in normal and abnormal sperm function. Frontiers in Bioscience. 1: 78-86.
23 - Sinha, M.P. 1996. The effect of glutathione on the motility, enzyme leakage and fertility of frozen goat semen, Animal Reproduction Science. 41: 237–243.
24- Stradaioli, G., T. Noro, L. Sylla and M. Monaci. 2007. Decrease in glutathione (GSH) content in bovine sperm after cryopreservation: Comparison between two extenders. Theriogenology. 67: 1249–1255.
25 -Tuncer, P.B., M.N. Bucak, S. Büyükleblebici, S. Sariözkan and et al. 2010. The effect of cysteine and glutathione on sperm and oxidative stress parameters of post-thawed bull semen. Cryobiology. 61: 303–307.
26 -Uysal, O. and M.N. Bucak. 2007. Effects of oxidized glutathione, bovine serum albumin, cysteine and lycopene on the quality of frozen-thawed ram semen. Acta Veterinaria Brno. 76: 383–390.