Abstract:
Fat (FAT atypical cadherin) and Dchs (Dachsous cadherin-related protein) in adjacent Sertoli:Sertoli, Sertoli:spermatid, and spermatid:spermatid interfaces create an important intercellular bridge whose adhesive function is in turn supported by Fjx1, a nonreceptor Ser/Thr protein kinase. This concept is derived from earlier studies of Drosophila, which has been confirmed in this and earlier reports as well. Herein, we use the approach of knockdown of Fat1 by RNAi using primary cultures of Sertoli cells that mimicked the blood-testis barrier (BTB) in vivo, and a series of coherent experiments including functional assays to monitor the Sertoli cell tight junction (TJ) permeability barrier and a functional in vitro TJ integrity assay to assess the role of Fat1 in the testis. It was shown that planar cell polarity (PCP) protein Fat1 affected Sertoli cell function through its modulation of actin and microtubule cytoskeletal function, altering their polymerization activity through the Fat1/Fjx1 complex. Furthermore, Fat1 is intimately associated with β-catenin and α-N-catenin, as well as with Prickle 1 of the Vangl1/Prickle 1 complex, another PCP core protein to support intercellular interactions to confer PCP. In summary, these findings support the notion that the Fat:Dchs and the Vangl2:Fzd PCP intercellular bridges are tightly associated with basal ES/TJ structural proteins to stabilize PCP function at the Sertoli:Sertoli, Sertoli:spermatid, and spermatid:spermatid interface to sustain spermatogenesis.
摘要:
邻近Sertoli中的脂肪(FAT非典型钙黏着蛋白)和Dchs(Dachsous钙黏着蛋白相关蛋白):Sertoli,Sertoli:精子,和精子细胞:精子细胞创造了一个重要的细胞间桥,其粘附功能又由非受体Ser/Thr蛋白激酶Fjx1支持。这个概念来自早期对果蝇的研究,这一点在这份报告和之前的报告中也得到了证实。在这里,我们使用RNAi敲低Fat1的方法,使用体内模拟血睾丸屏障(BTB)的支持细胞的原代培养物,以及一系列相关实验,包括监测支持细胞紧密连接(TJ)-通透性屏障的功能测定和评估Fat1在睾丸中的作用的功能体外TJ完整性测定。研究表明,平面细胞极性(PCP)蛋白Fat1通过调节肌动蛋白和微管细胞骨架功能来影响支持细胞功能,通过Fat1/Fjx1复合物改变其聚合活性。此外,Fat1与β-catenin和α-N-catenin密切相关,以及Vangl1/Prickle1复合体的Prickle1,另一种支持细胞间相互作用的PCP核心蛋白赋予PCP。总之,这些发现支持Fat:Dchs和Vangl2:FzdPCP细胞间桥与基础ES/TJ结构蛋白紧密相关,以稳定Sertoli:Sertoli的PCP功能,Sertoli:精子,和精子细胞:精子细胞界面维持精子发生。