Expression of peroxiredoxins in the human testis, epididymis and spermatozoa and their role in preventing H2O2-induced damage to spermatozoa
Abstract
Introduction. High levels of reactive oxygen species (ROS) have potential toxic effects on testicular function and sperm quality. Peroxiredoxins (PRDXs) are enzymes with a role as ROS scavenger. The aim of the study was to reveal the presence and localization of PRDXs in human testis, epididymis and spermatozoa, and the protective roles of PRDX2 and PRDX6 in sperm motility. Material and methods. The presence and localization of PRDXs in the human testis, epididymis and spermatozoa were detected by immunohistochemistry, western blot and immunofluorescence. The effect of anti-peroxidative damage to spermatozoa was examined by adding H2O2 to the recombinant protein-treated spermatozoa. Results. There were strong signals of PRDX1 in spermatogonia and round spermatids; PRDX2 in the round spermatids; PRDX4 and 5 in spermatogonia; PRDX6 in Sertoli cells. PRDXs were also found in epididymal epithelial cells where the expression of PRDX1, 4, 5, 6 in the cauda was higher than in the caput of epididymis. PRDX1-6 immunoreactivity was found throughout acrosome, post-acrosomal region, equatorial segment, neck and cytoplasmic droplet, midpiece and principal piece. The H2O2-induced reduction in sperm motility was reversed by recombinant PRDX2 or PRDX6 in a dose-dependent manner.
Conclusions. PRDX1-6 in the human testis and epididymis presented cell-specificity. PRDX2 and 6 are potential antioxidant protectors for human spermatozoa.
Keywords: Peroxiredoxinshumantestisepididymisspermatozoa
References
- Gagnon C, Iwasaki A, De Lamirande E, et al. Reactive oxygen species and human spermatozoa. Ann N Y Acad Sci. 1991; 637: 436–444.
- Garg A, Kumaresan A, Ansari MR. Effects of hydrogen peroxide (H2O2) on fresh and cryopreserved buffalo sperm functions during incubation at 37 degrees C in vitro. Reprod Domest Anim. 2009; 44(6): 907–912.
- Saleh R, Agarwal A, Kandirali E, et al. Leukocytospermia is associated with increased reactive oxygen species production by human spermatozoa. Fertility and Sterility. 2002; 78(6): 1215–1224.
- Sikka SC. Oxidative stress and role of antioxidants in normal and abnormal sperm function. Front Biosci. 1996; 1: e78–e86.
- Maneesh M, Jayalekshmi H, Dutta S, et al. Effect of chronic ethanol administration on testicular antioxidant system and steroidogenic enzyme activity in rats. Indian J Exp Biol. 2005; 43(5): 445–449.
- Padron OF, Brackett NL, Sharma RK, et al. Seminal reactive oxygen species and sperm motility and morphology in men with spinal cord injury. Fertil Steril. 1997; 67(6): 1115–1120.
- Gong S, San Gabriel MC, Zini A, et al. Low amounts and high thiol oxidation of peroxiredoxins in spermatozoa from infertile men. J Androl. 2012; 33(6): 1342–1351.
- Chae HZ, Kim HJ, Kang SW, et al. Characterization of three isoforms of mammalian peroxiredoxin that reduce peroxides in the presence of thioredoxin. Diabetes Res Clin Pract. 1999; 45(2-3): 101–112.
- Wood ZA, Schröder E, Robin Harris J, et al. Structure, mechanism and regulation of peroxiredoxins. Trends Biochem Sci. 2003; 28(1): 32–40.
- Murphy MP. How mitochondria produce reactive oxygen species. Biochem J. 2009; 417(1): 1–13.
- Anwar Sh, Yanai T, Sakai H. Overexpression of Peroxiredoxin 6 Protects Neoplastic Cells against Apoptosis in Canine Haemangiosarcoma. J Comp Pathol. 2016; 155(1): 29–39.
- Rahman MdS, Kwon WS, Lee JS, et al. Bisphenol-A affects male fertility via fertility-related proteins in spermatozoa. Sci Rep. 2015; 5: 9169.
- Sutovsky P, Aarabi M, Miranda-Vizuete A, et al. Negative biomarker based male fertility evaluation: Sperm phenotypes associated with molecular-level anomalies. Asian J Androl. 2015; 17(4): 554–560.
- Lee D, Moawad AR, Morielli T, et al. Peroxiredoxins prevent oxidative stress during human sperm capacitation. Mol Hum Reprod. 2017; 23(2): 106–115.
- Ozkosem B, Feinstein S, Fisher A, et al. Absence of Peroxiredoxin 6 Amplifies the Effect of Oxidant Stress on Mobility and SCSA/CMA3 Defined Chromatin Quality and Impairs Fertilizing Ability of Mouse Spermatozoa1. Biology of Reproduction. 2016; 94(3).
- Kwon WS, Rahman MdS, Lee JS, et al. A comprehensive proteomic approach to identifying capacitation related proteins in boar spermatozoa. BMC Genomics. 2014; 15: 897.
- Dammeyer P, Arnér ESJ. Human Protein Atlas of redox systems - what can be learnt? Biochim Biophys Acta. 2011; 1810(1): 111–138.
- Godoy J, Funke M, Ackermann W, et al. Redox atlas of the mouse. Biochimica et Biophysica Acta (BBA) - General Subjects. 2011; 1810(1): 2–92.
- Fujii T, Fujii J, Taniguchi N. Augmented expression of peroxiredoxin VI in rat lung and kidney after birth implies an antioxidative role. Eur J Biochem. 2001; 268(2): 218–225.
- O'Flaherty C, Souza AR. Hydrogen Peroxide Modifies Human Sperm Peroxiredoxins in a Dose-Dependent Manner. Biology of Reproduction. 2010; 84(2): 238–247.
- van Gestel RA, Brewis IA, Ashton PR, et al. Multiple proteins present in purified porcine sperm apical plasma membranes interact with the zona pellucida of the oocyte. Mol Hum Reprod. 2007; 13(7): 445–454.
- Shi H, Yu HJ, Wang HY, et al. Topical administration of peroxiredoxin-6 on the cornea suppresses inflammation and neovascularization induced by ultraviolet radiation. Invest Ophthalmol Vis Sci. 2012; 53(13): 8016–8028.
- Lin YQ, Li JY, Wang HY, et al. Cloning and identification of a novel sperm binding protein, HEL-75, with antibacterial activity and expressed in the human epididymis. Hum Reprod. 2008; 23(9): 2086–2094.
- Li J, Liu F, Liu X, et al. Mapping of the human testicular proteome and its relationship with that of the epididymis and spermatozoa. Mol Cell Proteomics. 2011; 10(3): M110.004630.
- Li J, Liu F, Wang H, et al. Systematic mapping and functional analysis of a family of human epididymal secretory sperm-located proteins. Mol Cell Proteomics. 2010; 9(11): 2517–2528.
- Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72: 248–254.
- Peltola V, Mäntylä E, Huhtaniemi I, et al. Lipid peroxidation and antioxidant enzyme activities in the rat testis after cigarette smoke inhalation or administration of polychlorinated biphenyls or polychlorinated naphthalenes. J Androl. 1994; 15(4): 353–361.
- Quinn PG, Payne AH. Oxygen-mediated damage of microsomal cytochrome P-450 enzymes in cultured leydig cells. Role in steroidogenic desensitization. J Biol Chem 1984; 259:4130-4135. ...nih. https://www ncbi nlm gov/pubmed/? term=J+Biol+Chem+1984%3B. ; 259: 3A4130–4135.
- Chen H, Liu J, Luo L, et al. Vitamin E, aging and Leydig cell steroidogenesis. Exp Gerontol. 2005; 40(8-9): 728–736.
- Bauché F, Fouchard MH, Jégou B. Antioxidant system in rat testicular cells. FEBS Lett. 1994; 349(3): 392–396.
- BOER PJ, POOT M, VERKERK A, et al. Glutathione-dependent defence mechanisms in isolated round spermatids from the rat. International Journal of Andrology. 1990; 13(1): 26–38.
- Aitken RJ, Curry BJ. Redox regulation of human sperm function: from the physiological control of sperm capacitation to the etiology of infertility and DNA damage in the germ line. Antioxid Redox Signal. 2011; 14(3): 367–381.
- Makker K, Agarwal A, Sharma R. Oxidative stress & male infertility. Indian J Med Res 2009; 129: 357-367. https://www.ncbi.nlm.nih.gov/pubmed/?term=Indian+J+Med+Res+2009%3B+129%3A+357-367. .
- Kodama H, Yamaguchi R, Fukuda J, et al. Increased oxidative deoxyribonucleic acid damage in the spermatozoa of infertile male patients. Fertility and Sterility. 1997; 68(3): 519–524.
- Manevich Y, Fisher AB. Peroxiredoxin 6, a 1-Cys peroxiredoxin, functions in antioxidant defense and lung phospholipid metabolism. Free Radic Biol Med. 2005; 38(11): 1422–1432.
- Nicolussi A, D'Inzeo S, Capalbo C, et al. The role of peroxiredoxins in cancer. Mol Clin Oncol. 2017; 6(2): 139–153.
- Barranco-Medina S, Lázaro JJ, Dietz KJ. The oligomeric conformation of peroxiredoxins links redox state to function. FEBS Lett. 2009; 583(12): 1809–1816.
- Tramer F, Rocco F, Micali F, et al. Antioxidant Systems in Rat Epididymal Spermatozoa1. Biology of Reproduction. 1998; 59(4): 753–758.
- Kropotov A, Usmanova N, Serikov V, et al. Mitochondrial targeting of human peroxiredoxin V protein and regulation of PRDX5 gene expression by nuclear transcription factors controlling biogenesis of mitochondria. FEBS J. 2007; 274(22): 5804–5814.
- Knoops B, Clippe A, Bogard C, et al. Cloning and characterization of AOEB166, a novel mammalian antioxidant enzyme of the peroxiredoxin family. J Biol Chem. 1999; 274(43): 30451–30458.
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