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.
Mol Med Rep. 2021 Feb;23(2). doi: 10.3892/mmr.2020.11791. Epub 2020 Dec 23.
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.
本研究旨在探讨三磷酸腺苷酶 Ca 转运质膜 4（PMCA4）在小鼠睾丸中的表达及其在精子发生中的作用。采用反转录-定量 PCR、western blot 和免疫荧光法评估 PMCA4 在野生型小鼠出生后不同周龄睾丸以及 Sertoli 细胞特异性雄激素受体敲除和雄激素受体敲除（ARKO）小鼠睾丸中的表达水平。通过荧光素酶检测、雄激素受体（AR）过表达和 AR 拮抗剂实验证实 AR 调节 PMCA4 的表达。结果表明，PMCA4 在出生后 3-8 周的小鼠睾丸中高度表达。与野生型相比，ARKO 小鼠睾丸中 PMCA4 的表达水平降低。此外，睾酮激活 AR 导致 PMCA4 启动子活性增加。转染 AR 过表达质粒的细胞中 PMCA4 蛋白的表达水平增加。最后，AR 拮抗剂氟他胺预处理可阻止睾酮诱导的 PMCA4 蛋白水平增加。本研究结果证实 PMCA4 在小鼠睾丸发育过程中上调，并且 PMCA4 mRNA 和蛋白表达水平受雄激素和 AR 调节。本研究结果提示 PMCA4 可能参与精子发生的调控。
