The Impact of NADPH Oxidase 5 Activity and Reactive Oxygen Species on Ca-pacitated Human Sperm

Hojjati Far, Mohammad; Keshtgar, Sara; Karbalaei, Narges

The Impact of NADPH Oxidase 5 Activity and Reactive Oxygen Species on Ca-pacitated Human Sperm

Vol. 25, Issue 3, / July-September 2024
(Original Article, pages 193-200)

Mohammad Hojjati Far: - Department of Physiology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
Sara Keshtgar: - Department of Physiology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
Narges Karbalaei: - Department of Physiology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran

Received: 4/24/2024 Accepted: 9/23/2024 - Publisher : Avicenna Research Institute

Other Format

Abstract

Background: Progesterone (P4) activates sperm calcium channels (CatSper), allowing calcium to enter the cell, which activates NADPH Oxidase-5 (NOX5) and produces reactive oxygen species (ROS). While calcium and ROS are essential for sperm capacitation, the role of NOX5 in capacitated sperm is unclear. This study investigated NOX5 activity in capacitated human sperm.
Methods: Forty semen samples from fertile men were processed, with motile sperm separated and divided into nine groups: control (Ham's F-10), solvent (DMSO), progesterone, diphenyleneiodonium chloride (DPI, NOX5 inhibitor), phorbol-12-myristate 13-acetate (PMA, NOX5 activator), P4+DPI, P4+PMA, Trolox, and P4+Trolox. Sperm kinematics, membrane integrity, survival rate, and ROS production was evaluated. Data were analyzed using ANOVA and Kruskal-Wallis tests, p£0.05 considered statistically significant.
Results: Progressive motility significantly decreased with DPI (56.2±2.1%) and PMA (56.5±2.1%), both alone and combined with progesterone (58.0±2.0% and 57.4±2.2%), compared to the progesterone group (66.0±1.9%). No significant change was observed in the Trolox groups. Progesterone, alone or combined with DPI, PMA, and Trolox, significantly reduced sperm linearity from 0.6±0.01 to 0.5±0.01%. Straightline velocity decreased in P4+PMA and P4+Trolox groups (88.2±4.4 and 89.7±3.9 μm/s) compared to the control group (105.0±5.5 μm/s). Trolox reduced ROS content, while other treatments had no effect on ROS levels.
Conclusion: NOX5 does not play a prominent role in capacitated sperm. The negative effects of PMA and DPI on sperm motility appear independent of their actions on NOX5 and ROS production. Trolox did not affect sperm motility and survival, indicating that capacitated sperm require little or no ROS.

Keywords: NADPH oxidase 5, Reactive oxygen species, Progesterone, Spermatozoa, Sperm capacitation, Trolox

To cite this article:

Hojjati Far M, Keshtgar S, Karbalaei N. The Impact of NADPH Oxidase 5 Activity and Reactive Oxygen Species on Ca-pacitated Human Sperm. J Reprod Infertil. 2024;25(3):193-200.

References

Lee KH, Hwang JY. Ca2+ homeostasis and male fertility: a target for a new male contraceptive system. Anim Cells Syst (Seoul). 2024;28(1):171-83. [PubMed]
Gagnon C, de Lamirande E. Controls of sperm motility. The sperm cell: production, maturation, fertilization, regeneration. Cambridge: Uk; 2006:108-33.
Puga Molina LC, Luque GM, Balestrini PA, Marin-Briggiler CI, Romarowski A, Buffone MG. Molecular basis of human sperm capacitation. Front Cell Dev Biol. 2018;6:72. [PubMed]
Delgado-Bermudez A, Yeste M, Bonet S, Pinart E. A review on the role of bicarbonate and proton transporters during sperm capacitation in mammals. Int J Mol Sci. 2022;23(11):6333. [PubMed]
Dai P, Zou M, Cai Z, Zeng X, Zhang X, Liang M. pH homeodynamics and male fertility: a coordinated regulation of acid-based balance during sperm journey to fertilization. Biomolecules. 2024;14(6):685. [PubMed]
Marín-Briggiler CI, Jha KN, Chertihin O, Buffone MG, Herr JC, Vazquez-Levin MnH, et al. Evidence of the presence of calcium/calmodulin-dependent protein kinase IV in human sperm and its involvement in motility regulation. J Cell Sci. 2005;118(Pt 9):2013-22. [PubMed]
O'Flaherty C. Redox regulation of mammalian sperm capacitation. Asian J Androl. 2015;17(4):583-90. [PubMed]
Naz RK, Rajesh PB. Role of tyrosine phosphorylation in sperm capacitation/acrosome reaction. Reprod Biol Endocrinol. 2004;2:75. [PubMed]
Luconi M, Francavilla F, Porazzi I, Macerola B, Forti G, Baldi E. Human spermatozoa as a model for studying membrane receptors mediating rapid nongenomic effects of progesterone and estrogens. Steroids. 2004;69(8-9):553-9. [PubMed]
Jiang F, Zhu Y, Chen Y, Tang X, Liu L, Chen G, et al. Progesterone activates the cyclic AMP-protein kinase a signalling pathway by upregul-ating ABHD2 in fertile men. J Int Med Res. 2021;49(3):300060521999527. [PubMed]
Musset B, Clark RA, DeCoursey TE, Petheo GL, Geiszt M, Chen Y, et al. NOX5 in human spermatozoa: expression, function, and regulation. J Biol Chem. 2012;287(12):9376-88. [PubMed]
Aitken RJ, Baker MA. The role of genetics and oxidative stress in the etiology of male infertility-a unifying hypothesis? Front Endocrinol (Lausanne). 2020;11:581838. [PubMed]
Chianese R, Pierantoni R. Mitochondrial reactive oxygen species (ROS) production alters sperm quality. Antioxidants (Basel). 2021;10(1):92. [PubMed]
Petrushanko IY, Lobachev VM, Kononikhin AS, Makarov AA, Devred F, Kovacic H, et al. Oxidation of Са2+-Binding Domain of NADPH Oxidase 5 (NOX5): Toward Understanding the Mechanism of Inactivation of NOX5 by ROS. PloS One. 2016;11(7):e0158726. [PubMed]
Touyz RM, Anagnostopoulou A, Rios F, Montezano AC, Camargo LL. NOX5: molecular biology and pathophysiology. Exp Physiol. 2019;104(5):605-16. [PubMed]
Keshtgar S, Ghani E. Impact of calcium and reactive oxygen species on human sperm function: role of NOX5. Andrologia. 2022;54(8):e14470. [PubMed]
Tirone F, Cox JA. NADPH oxidase 5 (NOX5) interacts with and is regulated by calmodulin. FEBS Lett. 2007;581(6):1202-8. [PubMed]
Chen F, Yu Y, Haigh S, Johnson J, Lucas R, Stepp DW, et al. Regulation of NADPH oxidase 5 by protein kinase C isoforms. PloS One. 2014;9(2):e88405. [PubMed]
Kirichok Y, Lishko PV. Rediscovering sperm ion channels with the patch-clamp technique. Mol Hum Reprod. 2011;17(8):478-99. [PubMed]
Strunker T, Goodwin N, Brenker C, Kashikar ND, Weyand I, Seifert R, et al. The CatSper channel mediates progesterone-induced Ca2+ influx in human sperm. Nature. 2011;471(7338):382-6. [PubMed]
Lishko PV, Botchkina I, Kirichok Y. Progesterone activates the principal Ca2+ channel of human sperm. Nature. 2011;471(7338):387-91. [PubMed]
Ghanbari H, Keshtgar S, Zare HR, Gharesi-Fard B. Inhibition of CatSper and Hv1 channels and NOX5 enzyme affect progesterone-induced increase of intracellular calcium concentration and ROS generation in human sperm. Iran J Med Sci. 2019;44(2):127-34. [PubMed]
Ghanbari H, Keshtgar S, Kazeroni M. Inhibition of the CatSper channel and NOX5 enzyme activity affects the functions of the progesterone-stimulated human sperm. Iran J Med Sci. 2018;43(1):18-25. [PubMed]
World Health O. WHO laboratory manual for the examination and processing of human semen. 6th ed. Geneva: World Health Organization; 2021. 290 p.
Jokiniemi A, Magris M, Ritari J, Kuusipalo L, Lundgren T, Partanen J, et al. Post-copulatory genetic matchmaking: HLA-dependent effects of cervical mucus on human sperm function. Proc Biol Sci. 2020;287(1933):20201682. [PubMed]
Keshtgar S, Ghanbari H, Ghani E, Shid Moosavi SM. Effect of CatSper and Hv1 channel inhibition on progesterone stimulated human sperm. J Reprod Infertil. 2018;19(3):133-9. [PubMed]
Goodson SG, White S, Stevans AM, Bhat S, Kao CY, Jaworski S, et al. CASAnova: a multiclass support vector machine model for the classification of human sperm motility patterns. Biol Reprod. 2017;97(5):698-708. [PubMed]
Miller MR, Mannowetz N, Iavarone AT, Safavi R, Gracheva EO, Smith JF, et al. Unconventional endocannabinoid signaling governs sperm activ-ation via the sex hormone progesterone. Science. 2016;352(6285):555-9. [PubMed]
Calogero AE, Burrello N, Barone N, Palermo I, Grasso U, D'Agata R. Effects of progesterone on sperm function: mechanisms of action. Hum Reprod. 2000;15 Suppl 1:28-45. [PubMed]
Armstrong JS, Bivalacqua TJ, Chamulitrat W, Sikka S, Hellstrom WJ. A comparison of the NADPH oxidase in human sperm and white blood cells. Int J Androl. 2002;25(4):223-9. [PubMed]
Teijeiro JM, Marini PE, Bragado MJ, Garcia-Marin LJ. Protein kinase C activity in boar sperm. Andrology. 2017;5(2):381-91. [PubMed]
Aitken RJ, Harkiss D, Knox W, Paterson M, Irvine DS. A novel signal transduction cascade in capacitating human spermatozoa characterised by a redox-regulated, cAMP-mediated induction of tyrosine phosphorylation. J Cell Sci. 1998;111(Pt 5):645-56. [PubMed]
Pullar JM, Hampton MB. Diphenyleneiodonium triggers the efflux of glutathione from cultured cells. J Biol Chem. 2002;277(22):19402-7. [PubMed]
Zavadskis S, Weidinger A, Hanetseder D, Banerjee A, Schneider C, Wolbank S, et al. Effect of diphenyleneiodonium chloride on intracellular reactive oxygen species metabolism with emphasis on NADPH oxidase and mitochondria in two therapeutically relevant human cell types. Pharmaceutics. 2020;13(1):10. [PubMed]
Ghani E, Keshtgar S, Habibagahi M, Ghannadi A, Kazeroni M. Expression of NOX5 in human teratozoospermia compared to normozoospermia. Andrologia. 2013;45(5):351-6. [PubMed]
Keshtgar S, Ebrahimi B, Shid-Moosavi SM, Erfani N. NADPH oxidase 5 activation; a novel approach to human sperm cryoinjury. Cell Tissue Bank. 2020;21(4):675-84. [PubMed]
Homa ST, Vessey W, Perez-Miranda A, Riyait T, Agarwal A. Reactive oxygen species (ROS) in human semen: determination of a reference range. J Assist Reprod Genet. 2015;32(5):757-64. [PubMed]
Ribas-Maynou J, Yeste M. Oxidative stress in male infertility: causes, effects in assisted reproductive techniques, and protective support of antioxidants. Biology (Basel). 2020;9(4):77. [PubMed]
Dutta S, Majzoub A, Agarwal A. Oxidative stress and sperm function: a systematic review on evaluation and management. Arab J Urol. 2019;17(2):87-97. [PubMed]
Villaverde AI, Netherton J, Baker MA. From past to present: the link between reactive oxygen species in sperm and male infertility. Antioxidants (Basel). 2019;8(12):616. [PubMed]
Cos P, Hermans N, Calomme M, Maes L, De Bruyne T, Pieters L, et al. Comparative study of eight well-known polyphenolic antioxidants. J Pharm Pharmacol. 2003;55(9):1291-7. [PubMed]
Keshtgar S, Iravanpour F, Gharesi-Fard B, Kazero-oni M. Combined effect of trolox and EDTA on frozen-thawed sperm quality. Iran J Med Sci. 2016;41(3):230-7. [PubMed]
Garrido N, Meseguer M, Simon C, Pellicer A, Re-mohi J. Pro-oxidative and anti-oxidative imbalance in human semen and its relation with male fertility. Asian J Androl. 2004;6(1):59-65. [PubMed]