Luoyang Normal University, Luoyang, 471934, Henan, China.
Cell Tissue Res. 2022 Jul;389(1):145-157. doi: 10.1007/s00441-022-03631-y. Epub 2022 May 10.
Communication between Sertoli cell is essential during spermatogenesis and testicular development in mice, and the dynamic balance of this communication is regulated by some adhesion proteins. In this study, we found that SPATA33 and CTNNA3 were involved in this process. Quantitative real-time PCR and western blotting showed similar trend of expression of two proteins in the testis of mice of different ages. Subsequently, CRISPR-Cas9 technique was used to prepare Spata33 knockout cell lines with TM4 cells, cell wound scratch assay showed that Spata33 gene knockout affected cell migration, and flow cytometry assay showed that Spata33 knockout resulted in a decreased percentage of G1 phase cells in TM4 cell line. In addition, phalloidin staining assay showed that Spata33 gene knockout disrupted the formation of F-actin. Moreover, the protein immunoprecipitation experiment showed the interaction between SPATA33 and CTNNA3, which affected the interaction between CTNNA3 and CTNNB1. SPATA33 inhibits the formation of CDH1-CTNNB1-CTNNA3 complex through its interaction with CTNNA3, thus weakening adhesion between Sertoli cell and promoting cell migration.
在精子发生和睾丸发育过程中，生精细胞之间的通讯对于精子发生和睾丸发育至关重要，这种通讯的动态平衡由一些黏附蛋白调节。在本研究中，我们发现 SPATA33 和 CTNNA3 参与了这一过程。定量实时 PCR 和 Western blot 显示，两种蛋白在不同年龄小鼠睾丸中的表达趋势相似。随后，我们使用 CRISPR-Cas9 技术制备了 Spata33 基因敲除 TM4 细胞系，细胞划痕实验表明 Spata33 基因敲除影响细胞迁移，流式细胞术实验表明 Spata33 基因敲除导致 TM4 细胞系中 G1 期细胞的百分比降低。此外，鬼笔环肽染色实验表明 Spata33 基因敲除破坏了 F-肌动蛋白的形成。此外，蛋白质免疫沉淀实验表明 SPATA33 与 CTNNA3 相互作用，从而影响 CTNNA3 与 CTNNB1 的相互作用。SPATA33 通过与 CTNNA3 的相互作用抑制 CDH1-CTNNB1-CTNNA3 复合物的形成，从而削弱了生精细胞之间的黏附，促进了细胞迁移。
