Browse > Article
http://dx.doi.org/10.5713/ab.21.0151

Cysteine improves boar sperm quality via glutathione biosynthesis during the liquid storage  

Zhu, Zhendong (College of Animal Science and Technology, Qingdao Agricultural University)
Zeng, Yao (Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University)
Zeng, Wenxian (Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University)
Publication Information
Animal Bioscience / v.35, no.2, 2022 , pp. 166-176 More about this Journal
Abstract
Objective: Sperm is particularly susceptible to reactive oxygen species (ROS) stress. Glutathione (GSH) is an endogenous antioxidant that regulates sperm redox homeostasis. However, it is not clear whether boar sperm could utilize cysteine for synthesis GSH to protect sperm quality from ROS damage. Therefore, the present study was undertaken to elucidate the mechanism of how cysteine is involved in protecting boar sperm quality during liquid storage. Methods: Sperm motility, membrane integrity, lipid peroxidation, 4-hydroxyIlonenal (4-HNE) modifications, mitochondrial membrane potential, as well as the levels of ROS, GSH, and, ATP were evaluated. Moreover, the enzymes (GCLC: glutamate cysteine ligase; GSS: glutathione synthetase) that are involved in glutathione synthesis from cysteine precursor were detected by western blotting. Results: Compared to the control, addition of 1.25 mM cysteine to the liquid storage significantly increased boar sperm progressive motility, straight-line velocity, curvilinear velocity, beat-cross frequency, membrane integrity, mitochondrial membrane potential, ATP level, acrosome integrity, activities of superoxide dismutase and catalase, and GSH level, while reducing the ROS level, lipid peroxidation and 4-HNE modifications. It was also observed that the GCLC and GSS were expressed in boar sperm. Interestingly, when we used menadione to induce sperm with ROS stress, the menadione associated damages were observed to be reduced by the cysteine supplementation. Moreover, compared to the cysteine treatment, the γ-glutamylcysteine synthetase (γ-GCS) activity, GSH level, mitochondrial membrane potential, ATP level, membrane integrity and progressive motility in boar sperm were decreased by supplementing with an inhibitor of GSH synthesis, buthionine sulfoximine. Conclusion: These data suggest that boar sperm could biosynthesize the GSH from cysteine in vitro. Therefore, during storage, addition of cysteine improves boar sperm quality via enhancing the GSH synthesis to resist ROS stress.
Keywords
Boar Sperm; Cysteine; Glutathione Synthesis; Reactive Oxygen Species Stress;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Zhu Z, Li R, Fan X, et al. Resveratrol improves boar sperm quality via 5'AMP-activated protein kinase activation during cryopreservation. Oxid Med Cell Longev 2019;2019:5921503. https://doi.org/10.1155/2019/5921503   DOI
2 Zhu Z, Li R, Wang L, et al. Glycogen synthase kinase-3 regulates sperm motility and acrosome reaction via affecting energy metabolism in goats. Front Physiol 2019;10:968. https://doi.org/10.3389/fphys.2019.00968   DOI
3 Murphy MP. How mitochondria produce reactive oxygen species. Biochem J 2009;417:1-13. https://doi.org/10.1016/j.freeradbiomed.2019.06.018   DOI
4 Mahfouz R, Sharma R, Thiyagarajan A, et al. Semen characteristics and sperm DNA fragmentation in infertile men with low and high levels of seminal reactive oxygen species. Fertil Steril 2010;94:2141-6. https://doi.org/10.1016/j.fertnstert.2009.12.030   DOI
5 Zhu Z, Fan X, Lv Y, et al. Vitamin E analogue improves rabbit sperm quality during the process of cryopreservation through its antioxidative action. PloS one 2015;10:e0145383. https://doi.org/10.1371/journal.pone.0145383   DOI
6 Zhu Z, Umehara T, Okazaki T, et al. Gene expression and protein synthesis in mitochondria enhance the duration of high-speed linear motility in boar sperm. Front Physiol 2019;10:252. https://doi.org/10.3389/fphys.2019.00252   DOI
7 Wang ST, Chen HW, Sheen LY, Lii CK. Methionine and cysteine affect glutathione level, glutathione-related enzyme activities and the expression of glutathione S-transferase isozymes in rat hepatocytes. J Nutr 1997;127:2135-41. https://doi.org/10.1093/jn/127.11.2135   DOI
8 Aitken RJ, Jones KT, Robertson SA. Reactive oxygen species and sperm function--in sickness and in health. J Androl 2012;33:1096-106. https://doi.org/10.2164/jandrol.112.016535   DOI
9 Aitken RJ. Reactive oxygen species as mediators of sperm capacitation and pathological damage. Mol Reprod Dev 2017;84:1039-52. https://doi.org/10.1002/mrd.22871   DOI
10 O'Donnell L, Nicholls PK, O'Bryan MK, McLachlan RI, Stanton PG. Spermiation: The process of sperm release. Spermatogenesis 2011;1:14-35. https://doi.org/10.4161/spmg.1.1.14525   DOI
11 Knox RV. Artificial insemination in pigs today. Theriogenology 2016;85:83-93. https://doi.org/10.1016/j.theriogenology.2015.07.009   DOI
12 Gadea J, Gumbao D, Gomez-Gimenez B, Gardon JC. Supplementation of the thawing medium with reduced glutathione improves function of frozen-thawed goat spermatozoa. Reprod Biol 2013;13:24-33. https://doi.org/10.1016/j.repbio.2013.01.174   DOI
13 Jan SZ, Hamer G, Repping S, de Rooij DG, van Pelt AMM, Vormer TL. Molecular control of rodent spermatogenesis. Biochim Biophys Acta Mol Basis Dis 2012;1822:1838-50. https://doi.org/10.1016/j.bbadis.2012.02.008   DOI
14 Kothari S, Thompson A, Agarwal A, du Plessis SS. Free radicals: their beneficial and detrimental effects on sperm function. Indian J Exp Biol 2010;48:425-35.
15 Zhu Z, Kawai T, Umehara T, MasudulHoque SA, Zeng W, Shimada M. Negative effects of ROS generated during linear sperm motility on gene expression and ATP generation in boar sperm mitochondria. Free Radic Biol Med 2019;141:159-71. https://doi.org/10.1016/j.freeradbiomed.2019.06.018   DOI
16 Gadea J, Selles E, Marco MA, et al. Decrease in glutathione content in boar sperm after cryopreservation: Effect of the addition of reduced glutathione to the freezing and thawing extenders. Theriogenology 2004;62:690-701. https://doi.org/10.1016/j.theriogenology.2003.11.013   DOI
17 Lu SC. Regulation of glutathione synthesis. Mol Aspects Med 2009;30:42-59. https://doi.org/10.1016/j.mam.2008.05.005   DOI
18 Ortega-Ferrusola C, Martin Munoz P, Ortiz-Rodriguez JM, et al. Depletion of thiols leads to redox deregulation, production of 4-hydroxinonenal and sperm senescence: a possible role for GSH regulation in spermatozoa. Biol Reprod 2019; 100:1090-107. https://doi.org/10.1093/biolre/ioy241   DOI
19 Birben E, Sahiner UM, Sackesen C, Erzurum S, Kalayci O. Oxidative stress and antioxidant defense. World Allergy Organ J 2012;5:9-19. https://doi.org/10.1097/WOX.0b013e3182439613   DOI
20 Gadea J, Gumbao D, Matas C, Romar R. Supplementation of the thawing media with reduced glutathione improves function and the in vitro fertilizing ability of boar spermatozoa after cryopreservation. J Androl 2005;26:749-56. https://doi.org/10.2164/jandrol.05057   DOI
21 Takeo T, Nakagata N. Reduced glutathione enhances fertility of frozen/thawed C57BL/6 mouse sperm after exposure to methyl-beta-cyclodextrin. Biol Reprod 2011;85:1066-72. https://doi.org/10.1095/biolreprod.111.092536   DOI
22 Lu SC. Glutathione synthesis. Biochim Biophys Acta Gen Subj 2013;1830:3143-53. https://doi.org/10.1016/j.bbagen.2012.09.008   DOI
23 Pagl R, Aurich C, Kankofer M. Anti-oxidative status and semen quality during cooled storage in stallions. J Vet Med A Physiol Pathol Clin Med 2006;53:486-9. https://doi.org/10.1111/j.1439-0442.2006.00879.x   DOI
24 Waberski D, Riesenbeck A, Schulze M, Weitze KF, Johnson L. Application of preserved boar semen for artificial insemination: Past, present and future challenges. Theriogenology 2019;137:2-7. https://doi.org/10.1016/j.theriogenology.2019.05.030   DOI
25 Lee AS, Lee SH, Lee S, Yang BK. Effects of streptozotocin and S-allyl-L-cysteine on motility, plasma membrane integrity, and mitochondrial activity of boar spermatozoa. Trop Anim Health Prod 2020;52:437-44. https://doi.org/10.1007/s11250-019-01983-2   DOI
26 Amaral S, Amaral A, Ramalho-Santos J. Aging and male reproductive function: a mitochondrial perspective. Front Biosci (Schol Ed) 2013;5:181-97. https://doi.org/10.2741/s365   DOI
27 Zhu Z, Li R, Feng C, et al. Exogenous oleic acid and palmitic acid improve boar sperm motility via enhancing mitochondrial beta-oxidation for ATP generation. Animals (Basel) 2020;10:591. https://doi.org/10.3390/ani10040591   DOI
28 Pena FJ, O'Flaherty C, Ortiz Rodriguez JM, et al. Redox regulation and oxidative stress: the particular case of the stallion spermatozoa. Antioxidants (Basel) 2019;8:567. https://doi.org/10.3390/antiox8110567   DOI
29 Aitken RJ, Smith TB, Jobling MS, Baker MA, De Iuliis GN. Oxidative stress and male reproductive health. Asian J Androl 2014;16:31-8. https://doi.org/10.4103/1008-682X.122203   DOI
30 Schulze M, Nitsche-Melkus E, Jakop U, Jung M, Waberski D. New trends in production management in European pig AI centers. Theriogenology 2019;137:88-92. https://doi.org/10.1016/j.theriogenology.2019.05.042   DOI
31 Salvador I, Yaniz J, Viudes-de-Castro MP, Gomez EA, Silvestre MA. Effect of solid storage on caprine semen conservation at 5 degrees C. Theriogenology 2006;66:974-81. https://doi.org/10.1016/j.theriogenology.2006.02.042   DOI
32 Tuncer PB, Bucak MN, Buyukleblebici S, et al. The effect of cysteine and glutathione on sperm and oxidative stress parameters of post-thawed bull semen. Cryobiology 2010;61:303-7. https://doi.org/10.1016/j.cryobiol.2010.09.009   DOI
33 Partyka A, Nizanski W, Bratkowska M, Maslikowski P. Effects of N-acetyl-L-cysteine and catalase on the viability and motility of chicken sperm during liquid storage. Reprod Biol 2015;15:126-9. https://doi.org/10.1016/j.repbio.2015.03.001   DOI
34 Fernandez M, O'Flaherty C, Moawad A, O'Flaherty C. Peroxiredoxins are key players of the enzymatic antioxidant system in human spermatozoa. Protein science. Hoboken, NJ, USA: Wiley; 2017. Vol. 26, pp. 165.
35 Zhu Z, Fan X, Lv Y, et al. Glutamine protects rabbit spermatozoa against oxidative stress via glutathione synthesis during cryopreservation. Reprod Fertil Dev 2017;29:2183-94. https://doi.org/10.1071/RD17020   DOI
36 Zhu Z, Ren Z, Fan X, et al. Cysteine protects rabbit spermatozoa against reactive oxygen species-induced damages. PLoS One 2017;12:e0181110. https://doi.org/10.1371/journal.pone.0181110   DOI
37 Gadea J, Molla M, Selles E, et al. Reduced glutathione content in human sperm is decreased after cryopreservation: Effect of the addition of reduced glutathione to the freezing and thawing extenders. Cryobiology 2011;62:40-6. https://doi.org/10.1016/j.cryobiol.2010.12.001   DOI
38 Bilodeau JF, Chatterjee S, Sirard MA, Gagnon C. Levels of antioxidant defenses are decreased in bovine spermatozoa after a cycle of freezing and thawing. Mol Reprod Dev 2000;55:282-8. https://doi.org/10.1002/(SICI)1098-2795(200003)55:3<282::AID-MRD6>3.0.CO;2-7   DOI