State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China
University of Chinese Academy of Sciences, Beijing, China
Reproduction. 2018 Oct 1;156(4):343–351. doi: 10.1530/REP-18-0148.
Reduced contractility of the testicular peritubular myoid (PTM) cells may contribute to human male subfertility or infertility. Transcription factor GATA4 in Sertoli and Leydig cells is essential for murine spermatogenesis, but limited attention has been paid to the potential role of GATA4 in PTM cells. In primary cultures of mouse PTM cells, siRNA knockdown of GATA4 increased the contractile activity, while GATA4 overexpression significantly attenuated the contractility of PTM cells using a collagen gel contraction assay. Using RNA sequencing and qRT-PCR, we identified a set of genes that exhibited opposite expressional alternation between Gata4 siRNA vs nontargeting siRNA-treated PTM cells and Gata4 adenovirus vs control adenovirus-treated PTM cells. Notably, ion channels, smooth muscle function, cytokines and chemokines, cytoskeleton, adhesion and extracellular matrix were the top four enriched pathways, as revealed by cluster analysis. Natriuretic peptide type B (NPPB) content was significantly upregulated by GATA4 overexpression in both PTM cells and their culture supernatant. More importantly, the addition of 100 μM NPPB could abolish the promoting effect of Gata4 silencing on PTM cell contraction. Taken together, we suggest that the inhibitory action of GATA4 on PTM cell contraction is mediated at least partly by regulating genes belonging to smooth muscle contraction pathway (e.g. Nppb).
睾丸小管周肌样细胞（PTM）收缩能力降低可能导致男性生育力下降或不育。Sertoli 和 Leydig 细胞中的转录因子 GATA4 对小鼠精子发生至关重要，但人们对 GATA4 在 PTM 细胞中的潜在作用关注有限。在原代培养的小鼠 PTM 细胞中，GATA4 的 siRNA 敲低增加了收缩活性，而使用胶原凝胶收缩测定，GATA4 的过表达显著减弱了 PTM 细胞的收缩性。通过 RNA 测序和 qRT-PCR，我们鉴定了一组基因，这些基因在 Gata4 siRNA 与非靶向 siRNA 处理的 PTM 细胞之间以及 Gata4 腺病毒与对照腺病毒处理的 PTM 细胞之间表现出相反的表达变化。值得注意的是，通过聚类分析发现，离子通道、平滑肌功能、细胞因子和趋化因子、细胞骨架、黏附和细胞外基质是前四个富集通路。GATA4 的过表达显著增加了 PTM 细胞及其培养上清液中脑钠肽 B（NPPB）的含量。更重要的是，添加 100μM 的 NPPB 可以消除 Gata4 沉默对 PTM 细胞收缩的促进作用。总之，我们认为 GATA4 对 PTM 细胞收缩的抑制作用至少部分是通过调节平滑肌收缩途径（如 Nppb）相关基因来介导的。
