Dong He-Ling, Wu Hong-Yuan, Fu You, Dai Meng, Bai Xiao-Chun, Wang Hong
College of Sports Science, Jinan University, Guangzhou 510632, China. E-mail:
Nan Fang Yi Ke Da Xue Xue Bao. 2017 Oct 20;37(10):1322-1329. doi: 10.3969/j.issn.1673-4254.2017.10.07.
To investigate the role of Rictor/mTORC2 in the formation of blood testis barrier (BTB), testicular development, and spermatogenesis.
Amh Cre positive mice homozygous for rictor loxP with Sertoli cell specific deletion of rictor were obtained by cross breeding Amh Cre mice with rictor loxP mice. The histology of the reproductive organs, seminiferous tubules and epididymis of the transgenic mice was observed with HE staining. The cell subgroups of the germ cells in the seminiferous tubule were detected by flow cytometry with propidium iodide labeling. The expression levels of Ki 67 and separase were detected with immunofluorescence assay, and the expression levels of BTB associated proteins were detected with immunofluorescence and Western blotting.
Compared with the control (Amh Cre, rictor or rictor) mice, the mice with Sertoli cell specific rictor deletion showed significantly decreased testicular weight and epididymis weight (P<0.05), significantly increased diploid cells (P<0.01), and decreased haploid cells (P<0.01) but comparable tetraploid cells and similar expression levels of Ki 67 and separase. The mice with rictor knockout also showed aberrant localization of BTB associated proteins, which were scattered over the whole seminiferous epithelium, but the expression levels of the protein remained stable.
Rictor in testicular Sertoli cells is essential for maintaining BTB integrity and function and ensuring normal spermatogenesis in mice.
探讨Rictor/mTORC2在血睾屏障（BTB）形成、睾丸发育及精子发生中的作用。
通过将Amh Cre小鼠与rictor loxP小鼠杂交，获得Sertoli细胞特异性敲除rictor的Amh Cre阳性且rictor loxP纯合的小鼠。用苏木精-伊红（HE）染色观察转基因小鼠生殖器官、生精小管和附睾的组织学变化。用碘化丙啶标记，通过流式细胞术检测生精小管中生殖细胞的细胞亚群。用免疫荧光法检测Ki 67和分离酶的表达水平，用免疫荧光和蛋白质印迹法检测BTB相关蛋白的表达水平。
与对照（Amh Cre、rictor或rictor）小鼠相比，Sertoli细胞特异性敲除rictor的小鼠睾丸重量和附睾重量显著降低（P<0.05），二倍体细胞显著增加（P<0.01），单倍体细胞减少（P<0.01），但四倍体细胞数量相当，Ki 67和分离酶的表达水平相似，并表现出BTB相关蛋白的异常定位，这些蛋白散布在整个生精上皮，但蛋白表达水平保持稳定。
睾丸Sertoli细胞中的Rictor对于维持小鼠BTB的完整性和功能以及确保正常精子发生至关重要。
