Expression of PPP3CC and not PPP3R2 is associated with asthenozoospermia
1Institute of Reproductive Medicine, Medical School, Nantong University, 226001 Nantong, China
2Institute of Life Science, Nanchang University, 330031 Nanchang, China
Submitted: 18 February 2021 Accepted: 10 March 2021
Online publish date: 20 May 2021
† These authors contributed equally.
Background and objective: Protein phosphatase 3 catalytic subunit gamma (PPP3CC) and protein phosphatase 3 regulatory subunit B, beta (PPP3R2) are respectively the catalytic and regulatory subunits of calcineurin in sperm. Deﬁciency in either protein causes impaired sperm motility leading to male infertility. Many cases of sterility are attributed to asthenozoospermia (AZS); however, it remains unknown whether PPP3CC and PPP3R2 are related to AZS.
Material and methods: Quantitative PCR and Western blotting were used to investigate the expression levels of PPP3CC and PPP3R2 in the spermatozoa of patients with AZS and to explore the clinical signiﬁcance.
Results: Two calcineurin inhibitors cyclosporine A (CsA) and tacrolimus (FK506) markedly impaired the total motility and progressive motility of human sperm, indicating that PPP3CC or PPP3R2 might be involved in AZS. PPP3CC mRNA and protein expression was lower in the ejaculated spermatozoa of patients with AZS than in normal sperm (NS). Correlation analysis showed that PPP3CC protein expression correlated positively with progressive motility (r = 0.2592, P < 0.05); however, there were no signiﬁcant differences in PPP3R2 mRNA and protein levels between AZS and NS.
Conclusion: These ﬁndings suggest that the abnormal expression of PPP3CC rather than PPP3R2 might be a pathological factor or indicator in AZS. Thus, PPP3CC may be a potential therapeutic or diagnostic target for some cases of male infertility.
Asthenozoospermia; Calcineurin; PPP3CC; PPP3R2; Sperm motility; Progressive motility
Hang Kang,Chen Chen,Feng-Xin Gao,Xu-Hui Zeng,Xiao-Ning Zhang. Expression of PPP3CC and not PPP3R2 is associated with asthenozoospermia. Journal of Men's Health. 2021.doi:10.31083/jomh.2021.039.
 Feng HL. Molecular biology of male infertility. Archives of Andrology. 2003; 49: 19–27.
 Moore FL, Reijo-Pera RA. Male sperm motility dictated by mother’s mtDNA. American Journal of Human Genetics. 2000; 67: 543–548.
 Curi SM, Ariagno JI, Chenlo PH, Mendeluk GR, Pugliese MN, Sardi Segovia LM, et al. Asthenozoospermia: analysis of a large population. Archives of Andrology. 2003; 49: 343–349.
 Cooper TG, Noonan E, von Eckardstein S, Auger J, Baker HWG, Behre HM, et al. World Health Organization reference values for human semen characteristics. Human Reproduction Update. 2010; 16: 231–245.
 Zhang X, Diao R, Zhu X, Li Z, Cai Z. Metabolic characterization of asthenozoospermia using nontargeted seminal plasma metabolomics. Clinica Chimica Acta. 2015; 450: 254–261.
 Yang Y, Cheng L, Wang Y, Han Y, Liu J, Deng X, et al. Expression of NDUFA13 in asthenozoospermia and possible pathogenesis. Repro-ductive BioMedicine Online. 2017; 34: 66–74.
 Shen S, Wang J, Liang J, Zhu C. Low-expressed testis-specific calcium-binding protein CBP86-IV (CABYR) is observed in idiopathic asthenozoospermia. World Journal of Urology. 2015; 33: 633–638.
 Cheng Y, Hu X, Peng Z, Pan T, Wang F, Chen H, et al. Lysine glutarylation in human sperm is associated with progressive motility. Human Reproduction. 2019; 34: 1186–1194.
 Borghei A, Ouyang Y, Westmuckett AD, Marcello MR, Landel CP, Evans JP, et al. Targeted disruption of tyrosylprotein sulfotransferase-2, an enzyme that catalyzes post-translational protein tyrosine O-sulfation, causes male infertility. Journal of Biological Chemistry. 2006; 281: 9423–9431.
 Gunes S, Arslan MA, Hekim GNT, Asci R. The role of epigenetics in idiopathic male infertility. Journal of Assisted Reproduction and Genetics. 2017; 33: 553–569.
 Holyoake AJ, McHugh P, Wu M, O’Carroll S, Benny P, Sin IL, et al. High incidence of single nucleotide substitutions in the mitochondrial genome is associated with poor semen parameters in men. Interna-tional Journal of Andrology. 2001; 24: 175–182.
 Pereira L, Gonçalves J, Bandelt H. Mutation C11994T in the mitochondrial ND4 gene is not a cause of low sperm motility in Portugal. Fertility and Sterility. 2008; 89: 738–741.
 Zhang L, Liu Z, Li X, Zhang P, Wang J, Zhu D, et al. Low long non-coding RNA HOTAIR expression is associated with down-regulation of Nrf2 in the spermatozoa of patients with asthenozoospermia or oligoasthenozoospermia. International Journal of Clinical and Experimental Pathology. 2015; 8: 14198–14205.
 Zhou R, Wang R, Qin Y, Ji J, Xu M, Wu W, et al. Mitochondria-related miR-151a-5p reduces cellular ATP production by targeting CYTB in asthenozoospermia. Scientific Reports. 2016; 5: 17743.
 Zhou J, Zhou Q, Lyu X, Zhu T, Chen Z, Chen M, et al. The expression of cysteine-rich secretory protein 2 (CRISP2) and its specific regulator miR-27b in the spermatozoa of patients with asthenozoospermia. Biology of Reproduction. 2015; 92: 28.
 Wang L, Chen L, Liu S, Li Y, Zou L, Guan Y, et al. Chloride channels are involved in sperm motility and are downregulated in spermatozoa from patients with asthenozoospermia. Asian Journal of Andrology. 2017; 19: 418.
 Heidary Z, Zaki-Dizaji M, Saliminejad K, Khorramkhorshid HR. Expression analysis of the CRISP2, CATSPER1, PATE1 and SEMG1 in the sperm of men with idiopathic asthenozoospermia. Journal of Reproduction & Infertility. 2020; 20: 70–75.
 An C, Jiang H, Wang Q, Yuan R, Liu J, Shi W, et al. Down-regulation of DJ-1 protein in the ejaculated spermatozoa from Chinese asthenozoospermia patients. Fertility and Sterility. 2011; 96: 19–23.e2.
 Gooch JL, Gorin Y, Zhang B, Abboud HE. Involvement of calcineurin in transforming growth factor-beta-mediated regulation of extracel-lular matrix accumulation. Journal of Biological Chemistry. 2004; 279: 15561–15570.
 Parra V, Rothermel BA. Calcineurin signaling in the heart: the importance of time and place. Journal of Molecular and Cellular Cardiology. 2017; 103: 121–136.
 Muramatsu T, Giri PR, Higuchi S, Kincaid RL. Molecular cloning of a calmodulin-dependent phosphatase from murine testis: identification of a developmentally expressed nonneural isoenzyme. Proceedings of the National Academy of Sciences of the United States of America. 1992; 89: 529–533.
 Means AR, Tash JS, Chafouleas JG. Physiological implications of the presence, distribution, and regulation of calmodulin in eukaryotic cells. Physiological Reviews. 1982; 62: 1–39.
 Moriya M, Katagiri C, Yagi K. Immuno-electron microscopic local-ization of calmodulin and calmodulin-binding proteins in the mouse germ cells during spermatogenesis and maturation. Cell and Tissue Research. 1993; 271: 441–451.
 Parte PP, Rao P, Redij S, Lobo V, D’Souza SJ, Gajbhiye R, et al. Sperm phosphoproteome profiling by ultra performance liquid chromatography followed by data independent analysis (LC-MS(E)) reveals altered proteomic signatures in asthenozoospermia. Journal of Proteomics. 2013; 75: 5861–5871.
 Cao Z, Liu J, Zhu Y, Zhou S, Qi L, Dong X, et al. Effects of different immunodepressants on the sperm parameters of kidney transplant recipients. Zhonghua Nan Ke Xue. 2006; 12: 405–407. (In Chinese)
 Xu LG, Xu YH, Xu HM, Zhu XF, Ling X, Zhang JR, et al. The effect of cyclosporine A on semen quality in kidney transplant patients. Journal of Clinical Urology. 2005; 20, 603–605.
 Misro MM, Chaki SP, Srinivas M, Chaube SK. Effect of cyclosporine on human sperm motility in vitro. Archives of Andrology. 1999; 43: 215–220.
 Miyata H, Satouh Y, Mashiko D, Muto M, Nozawa K, Shiba K, et al. Sperm calcineurin inhibition prevents mouse fertility with implications for male contraceptive. Science. 2015; 350: 442–445.
 Zhang X, Zhang P, Song D, Xiong S, Zhang H, Fu J, et al. Expression profiles and characteristics of human lncRNA in normal and asthenozoospermia sperm. Biology of Reproduction. 2019; 100: 982–993.
 Tash JS, Krinks M, Patel J, Means RL, Klee CB, Means AR. Identi-fication, characterization, and functional correlation of calmodulin-dependent protein phosphatase in sperm. Journal of Cell Biology. 1988; 106: 1625–1633.
 Liu Y, Zhang C, Wang S, Hu Y, Jing J, Ye L, et al. Dependence of sperm structural and functional integrity on testicular calcineurin isoform PPP3R2 expression. Journal of Molecular Cell Biology. 2020; 12: 515–529.
 Hisatomi A, Fujihira S, Fujimoto Y, Fujii T, Mine Y, Ohara K. Effect of Prograf (FK506) on spermatogenesis in rats. Toxicology. 1996; 109: 75–83.
 Heidary Z, Zaki‐Dizaji M, Saliminejad K, Khorram Khorshid HR. MicroRNA profiling in spermatozoa of men with unexplained asthenozoospermia. Andrologia. 2019; 51: e13284.
 Jodar M, Kalko S, Castillo J, Ballescà JL, Oliva R. Differential RNAs in the sperm cells of asthenozoospermic patients. Human Reproduction. 2012; 27: 1431–1438.
 Zhou J, Zhou Q, Yang J, Lyu X, Bian J, Guo W, et al. MicroRNA-27a-mediated repression of cysteine-rich secretory protein 2 translation in asthenoteratozoospermic patients. Asian Journal of Andrology. 2017; 19: 591–595.
 Carrera A, Moos J, Ning XP, Gerton GL, Tesarik J, Kopf GS, et al. Regulation of protein tyrosine phosphorylation in human sperm by a calcium/calmodulin-dependent mechanism: identification of a kinase anchor proteins as major substrates for tyrosine phosphorylation. Developmental Biology. 1996; 180: 284–296.
 Chan C, Shui H, Wu C, Wang C, Sun G, Chen H, et al. Motility and protein phosphorylation in healthy and asthenozoospermic sperm. Journal of Proteome Research. 2009; 8: 5382–5386.
 Vadnais ML, Aghajanian HK, Lin A, Gerton GL. Signaling in sperm: toward a molecular understanding of the acquisition of sperm motility in the mouse epididymis. Biology of Reproduction. 2013; 89: 127.
 Espino J, Mediero M, Lozano GM, Bejarano I, Ortiz Á, García JF, et al. Reduced levels of intracellular calcium releasing in spermatozoa from asthenozoospermic patients. Reproductive Biology and Endocrinol-ogy. 2009; 7: 1–11.
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