Department of Urology, Affiliated Shenzhen Longhua People's Hospital, Southern Medical University 518109, Guangdong, China.
Central Laboratory, Affiliated Shenzhen Longhua People's Hospital, Southern Medical University 518109, Guangdong, China.
Syst Biol Reprod Med. 2022 Feb;68(1):70-79. doi: 10.1080/19396368.2021.2004471. Epub 2021 Dec 12.
Androgen and AR are essential for maintaining spermatogenesis and male fertility. Previous studies have shown that the phosphatidyl ethanolamine binding protein 1 () gene is down-regulated in the selective ablation of the AR in the Sertoli cells of mouse testes compared with wild-type mice, indicating that is a candidate target of AR. The ChIP-PCR data and ChIP-sequencing results of this study verified that is a target gene regulated by AR. Real-time PCR, Western blot analysis, and immunofluorescence data showed that is expressed at all stages of testicular development, with an increasing trend from 1 to 8 weeks of postnatal development. PEBP1 was principally located in the cytoplasm, and high-intensity fluorescence revealed PEBP in the lumen of the testicular tubules. Bioinformatics analysis indicated effective androgen-responsive elements (AREs) located in the promotor of gene. Dual fluorescence assay data showed that androgens and AR could bind to the AREs of and induce an increase of gene expression. These data suggest that is a newfound target gene regulated by androgens and AR in mouse Sertoli cells. However, the detailed molecular mechanism of their role in spermatogenesis still needs to be further studied. AR: androgen receptor; Pebp1: phosphatidyl ethanolamine binding protein 1; ARKO: androgen receptor knockout; WT: wild type; SCARKO: Sertoli cell-selective androgen receptor knockout; ChIP: chromatin immunoprecipitation; RKIP: Raf kinase inhibitory protein; MAPK: mitogen-activated protein kinase; NF-κB: nuclear factor kappa-light-chain-enhancer of activated B cells; GSK-3: glycogen synthase kinase-3; RT-PCR: reverse transcriptase polymerase chain reaction; SEM: standard error of the mean.
雄激素和 AR 对于维持精子发生和男性生育能力至关重要。先前的研究表明，与野生型小鼠相比，在小鼠睾丸支持细胞中 AR 的选择性缺失中，磷酸乙醇胺结合蛋白 1 () 基因下调，表明 是 AR 的候选靶标。本研究的 ChIP-PCR 数据和 ChIP-seq 结果验证了 是受 AR 调节的靶基因。实时 PCR、Western blot 分析和免疫荧光数据表明，在睾丸发育的所有阶段都表达 ，从出生后 1 周到 8 周呈上升趋势。PEBP1 主要位于细胞质中，高强度荧光显示睾丸小管管腔中的 PEBP。生物信息学分析表明，位于 基因启动子中的有效雄激素反应元件 (AREs)。双荧光测定数据表明，雄激素和 AR 可以与 的 AREs 结合并诱导基因表达增加。这些数据表明，是小鼠支持细胞中受雄激素和 AR 调节的新发现的靶基因。然而，它们在精子发生中的作用的详细分子机制仍需要进一步研究。AR：雄激素受体；Pebp1：磷酸乙醇胺结合蛋白 1；ARKO：雄激素受体敲除；WT：野生型；SCARKO：支持细胞选择性雄激素受体敲除；ChIP：染色质免疫沉淀；RKIP：Raf 激酶抑制剂蛋白；MAPK：丝裂原激活蛋白激酶；NF-κB：核因子 kappa-轻链增强子的 B 细胞；GSK-3：糖原合酶激酶-3；RT-PCR：逆转录聚合酶链反应；SEM：平均值的标准误差。
