PMCA4 gene expression is regulated by the androgen receptor in the mouse testis during spermatogenesis

Sun,Rui; Liang,Hui; Guo,Huan; Wang,Zhu; Deng,Qiong

doi:10.3892/mmr.2020.11791

PMCA4 gene expression is regulated by the androgen receptor in the mouse testis during spermatogenesis

Authors:
- Rui Sun
- Hui Liang
- Huan Guo
- Zhu Wang
- Qiong Deng
View Affiliations

Affiliations: Department of Urology, The People's Hospital of Longhua, The Affiliated Hospital of Southern Medical University, Shenzhen, Guangdong 518109, P.R. China, Department of Urology, Shenzhen University General Hospital and Shenzhen University Clinical Medical Academy Center, Shenzhen University, Shenzhen, Guangdong 518052, P.R. China
Published online on: December 20, 2020 https://doi.org/10.3892/mmr.2020.11791
Article Number: 152
Copyright: © Sun et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The present study aimed to investigate the expression of ATPase Ca⁺⁺ transporting plasma membrane 4 (PMCA4) in mouse testis and to determine its role in spermatogenesis. Reverse transcription‑quantitative PCR, western blotting and immunofluorescence were performed to evaluate the expression levels of PMCA4 in mouse testes at various weeks postnatal in wild type mice, and in testes from Sertoli cell‑specific androgen receptor knockout and androgen receptor knockout (ARKO) mice. Luciferase assay, androgen receptor (AR) overexpression and AR antagonist experiments were used to confirm that AR regulated the expression of PMCA4. The results demonstrated that PMCA4 was highly expressed in mouse testes at 3‑8 weeks postnatal. PMCA4 expression levels in ARKO mouse testes were decreased compared with wild type. In addition, activation of AR by testosterone administration resulted in an increase in the activity of the PMCA4 promoter. Cells transfected with an AR‑overexpressing plasmid exhibited increased expression levels of the PMCA4 protein. Finally, the increase in PMCA4 protein levels induced by testosterone was prevented by pre‑treatment with the AR antagonist flutamide. The present results confirmed that PMCA4 was upregulated during mouse testis development and that PMCA4 mRNA and protein expression levels were regulated by androgens and AR. The present findings suggest that PMCA4 may be involved in the regulation of spermatogenesis.

View Figures

View References

1	Walker WH: Testosterone signaling and the regulation of spermatogenesis. Spermatogenesis. 1:116–120. 2011. View Article : Google Scholar : PubMed/NCBI
2	Yong EL, Lim LS, Wang Q, Mifsud A, Lim J, Ong YC and Sim KS: Androgen receptor polymorphisms and mutations in male infertility. J Endocrinol Invest. 23:573–577. 2000. View Article : Google Scholar : PubMed/NCBI
3	Tahmasbpour E, Balasubramanian D and Agarwal A: A multi-faceted approach to understanding male infertility: Gene mutations, molecular defects and assisted reproductive techniques (ART). J Assist Reprod Genet. 31:1115–1137. 2014. View Article : Google Scholar : PubMed/NCBI
4	O'Hara L and Smith LB: Androgen receptor roles in spermatogenesis and infertility. Best Pract Res Clin Endocrinol Metab. 29:595–605. 2015. View Article : Google Scholar : PubMed/NCBI
5	De Gendt K, Swinnen JV, Saunders PT, Schoonjans L, Dewerchin M, Devos A, Tan K, Atanassova N, Claessens F, Lécureuil C, et al: A Sertoli cell-selective knockout of the androgen receptor causes spermatogenic arrest in meiosis. Proc Natl Acad Sci USA. 101:1327–1332. 2004. View Article : Google Scholar : PubMed/NCBI
6	MacLean JA II and Wilkinson MF: The Rhox genes. Reproduction. 140:195–213. 2010. View Article : Google Scholar : PubMed/NCBI
7	Zhang QX, Zhang XY, Zhang ZM, Lu W, Liu L, Li G, Cai ZM, Gui YT and Chang C: Identification of testosterone-/androgen receptor-regulated genes in mouse Sertoli cells. Asian J Androl. 14:294–300. 2012. View Article : Google Scholar : PubMed/NCBI
8	Calì T, Brini M and Carafoli E: Regulation of cell calcium and role of plasma membrane calcium ATPases. Int Rev Cell Mol Biol. 332:259–296. 2017. View Article : Google Scholar : PubMed/NCBI
9	Schuh K, Cartwright EJ, Jankevics E, Bundschu K, Liebermann J, Williams JC, Armesilla AL, Emerson M, Oceandy D, Knobeloch KP, et al: Plasma membrane Ca²⁺ ATPase 4 is required for sperm motility and male fertility. J Biol Chem. 279:28220–28226. 2004. View Article : Google Scholar : PubMed/NCBI
10	Patel R, Al-Dossary AA, Stabley DL, Barone C, Galileo DS, Strehler EE and Martin-DeLeon PA: Plasma membrane Ca²⁺-ATPase 4 in murine epididymis: Secretion of splice variants in the luminal fluid and a role in sperm maturation. Biol Reprod. 89:62013. View Article : Google Scholar : PubMed/NCBI
11	Olli KE, Li K, Galileo DS and Martin-DeLeon PA: Plasma membrane calcium ATPase 4 (PMCA4) co-ordinates calcium and nitric oxide signaling in regulating murine sperm functional activity. J Cell Physiol. 233:11–22. 2018. View Article : Google Scholar : PubMed/NCBI
12	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
13	Ma Q, Li Y, Luo M, Guo H, Lin S, Chen J, Du Y, Jiang Z and Gui Y: The expression characteristics of FAM71D and its association with sperm motility. Hum Reprod. 32:2178–2187. 2017. View Article : Google Scholar : PubMed/NCBI
14	Deng Q, Zhang Z, Wu Y, Yu WY, Zhang J, Jiang ZM, Zhang Y, Liang H and Gui YT: Non-genomic action of androgens is mediated by rapid phosphorylation and regulation of androgen receptor trafficking. Cell Physiol Biochem. 43:223–236. 2017. View Article : Google Scholar : PubMed/NCBI
15	Vignini A, Buldreghini E, Nanetti L, Amoroso S, Boscaro M, Ricciardo-Lamonica G, Mazzanti L and Balercia G: Free thiols in human spermatozoa: Are Na⁺/K⁺-ATPase, Ca²⁺-ATPase activities involved in sperm motility through peroxynitrite formation? Reprod Biomed Online. 18:132–140. 2009. View Article : Google Scholar : PubMed/NCBI
16	Jimenez T, Sánchez G and Blanco G: Activity of the Na,K-ATPase α4 isoform is regulated during sperm capacitation to support sperm motility. J Androl. 33:1047–1057. 2012. View Article : Google Scholar : PubMed/NCBI
17	Wood CD, Darszon A and Whitaker M: Speract induces calcium oscillations in the sperm tail. J Cell Biol. 161:89–101. 2003. View Article : Google Scholar : PubMed/NCBI
18	Fukami K, Yoshida M, Inoue T, Kurokawa M, Fissore RA, Yoshida N, Mikoshiba K and Takenawa T: Phospholipase Cdelta4 is required for Ca²⁺ mobilization essential for acrosome reaction in sperm. J Cell Biol. 161:79–88. 2003. View Article : Google Scholar : PubMed/NCBI
19	Rahman F and Christian HC: Non-classical actions of testosterone: An update. Trends Endocrinol Metab. 18:371–378. 2007. View Article : Google Scholar : PubMed/NCBI
20	Smith LB and Walker WH: The regulation of spermatogenesis by androgens. Semin Cell Dev Biol. 30:2–13. 2014. View Article : Google Scholar : PubMed/NCBI
21	Calamera J, Buffone M, Ollero M, Alvarez J and Doncel GF: Superoxide dismutase content and fatty acid composition in subsets of human spermatozoa from normozoospermic, asthenozoospermic, and polyzoospermic semen samples. Mol Reprod Dev. 66:422–430. 2003. View Article : Google Scholar : PubMed/NCBI
22	Wilhelm B, Brandenburger T, Post H and Aumüller G: Expression and localization of PMCA4 in rat testis and epididymis. Histochem Cell Biol. 129:331–343. 2008. View Article : Google Scholar : PubMed/NCBI
23	Di Leva F, Domi T, Fedrizzi L, Lim D and Carafoli E: The plasma membrane Ca2+ ATPase of animal cells: Structure, function and regulation. Arch Biochem Biophys. 476:65–74. 2008. View Article : Google Scholar : PubMed/NCBI
24	Okunade GW, Miller ML, Pyne GJ, Sutliff RL, O'Connor KT, Neumann JC, Andringa A, Miller DA, Prasad V, Doetschman T, et al: Targeted ablation of plasma membrane Ca2+-ATPase (PMCA) 1 and 4 indicates a major housekeeping function for PMCA1 and a critical role in hyperactivated sperm motility and male fertility for PMCA4. J Biol Chem. 279:33742–33750. 2004. View Article : Google Scholar : PubMed/NCBI
25	Chen Y, Qi C, Xia L and Li G: Identification of novel genetic etiology and key molecular pathways for seminoma via network-based studies. Int J Oncol. 51:1280–1290. 2017. View Article : Google Scholar : PubMed/NCBI
26	Margolin G, Khil PP, Kim J, Bellani MA and Camerini-Otero RD: Integrated transcriptome analysis of mouse spermatogenesis. BMC Genomics. 15:392014. View Article : Google Scholar : PubMed/NCBI
27	Brandenburger T, Strehler EE, Filoteo AG, Caride AJ, Aumüller G, Post H, Schwarz A and Wilhelm B: Switch of PMCA4 splice variants in bovine epididymis results in altered isoform expression during functional sperm maturation. J Biol Chem. 286:7938–7946. 2011. View Article : Google Scholar : PubMed/NCBI
28	Deng Q, Wu Y, Zhang Z, Wang Y, Li M, Liang H and Gui Y: Androgen receptor localizes to plasma membrane by binding to caveolin-1 in mouse sertoli cells. Int J Endocrinol. 2017:39859162017. View Article : Google Scholar : PubMed/NCBI
29	Wang RS, Yeh S, Tzeng CR and Chang C: Androgen receptor roles in spermatogenesis and fertility: Lessons from testicular cell-specific androgen receptor knockout mice. Endocr Rev. 30:119–132. 2009. View Article : Google Scholar : PubMed/NCBI
30	Lestari SW, Miati DN, Seoharso P, Sugiyanto R and Pujianto DA: Sperm Na⁺, K⁺-ATPase α4 and plasma membrane Ca²⁺-ATPase (PMCA) 4 regulation in asthenozoospermia. Syst Biol Reprod Med. 63:294–302. 2017. View Article : Google Scholar : PubMed/NCBI