RC/BTB2对纤毛形成及精子发生的作用

Cilia are conserved organelles that project from cell surface, they exist in almost all mammalian cells. There are two general types of cilia, primary cilia and motile cilia. Recent studies demonstrated that primary cilia are involved in several signal pathways, and they play an essential role in the embryonic development. The major functions of motile cilia are to regulate sperm motility, remove pathogens from respiratory tract, and circulate Cerebro-Spinal fluid. Defect in ciliary function gives rise to many genetic diseases, including male infertility, polycystic kidney, Bardet-Biedl Syndrome, hydrocephalus, infection in upper respiratory tract et al. RC/BTB2 is a protein that interacts with sperm association antigen 16 (SPAG16), a key regulator for sperm flagella formation and motility. In the testis, the protein is present in all the structures that are involved in sperm flagella formation, such as Golgi body, manchette, centrioles of germ cells, and is expressed in mature sperm flagella. In somatic cells, it is also present in structures related in ciliogenesis, including the Golgi bodies and centrioles, thus, defect in RC/BTB2 might be a major cause for ciliopathies. The purpose of this project is to study the role of this gene in ciliogenesis in vivo by comprehensive techniques, including gene targeting. This study will help to dissect novel mechanism of ciliogenesis and spermatogenesis, and provide foundation to treat the disease.

纤毛是一种突出于细胞表面且结构保守的细胞器，分原纤毛和运动纤毛，其中原纤毛主要参与胚胎发育和器官形成，运动纤毛主要参与精子鞭毛运动，支气管异物清除及脑脊液循环等。纤毛结构和功能障碍将导致胚胎发育不良、器官形成缺陷、男性不孕、呼吸系统炎症、脑水肿等多种疾病。RC/BTB2蛋白与精子相关抗原16相互作用，该蛋白不仅在与精子鞭毛形成相关结构如高尔基体、精子领、中心体中表达，且在可形成原纤毛的小鼠成纤维细胞和肾内髓集合管3上皮细胞中定位于调节纤毛形成的重要细胞器，提示该蛋白可能同时调节原纤毛和运动纤毛形成，该基因功能异常可能是导致纤毛病的重要原因。本研究将在小鼠整体水平，结合基因敲除等手段，系统研究观察RC/BTB2对纤毛形成和精子发生的影响，为纤毛病发生机制和防治药物开发提供理论依据。

纤毛分原纤毛和运动纤毛，原纤毛主要参与胚胎发育和器官形成，而运动纤毛主要参与精子鞭毛运动，支气管异物清除及脑脊液循环等。纤毛结构和功能障碍将导致胚胎发育不良、器官形成缺陷、男性不孕、呼吸系统炎症、脑水肿等多种疾病。Spag16调节精子发生，而RC/BTB2蛋白与spag16相互作用，所以RC/BTB2可能参与精子运动及纤毛形成。本研究在细胞水平和小鼠整体水平，结合酵母双杂交，RNA干扰，基因敲除等手段，全面系统的研究RC/BTB2对纤毛形成和精子发生的影响。结果显示，在小鼠睾丸中，RC/BTB2蛋白不仅在与精子鞭毛形成相关结构如高尔基体、精子领、中心体中表达，而且在NIH3T3细胞和IMCD3细胞中定位于调节纤毛形成的重要细胞器（高尔基体及中心体），RC/BTB2 RNA干扰可显著减少细胞纤毛的形成。总之，我们的结果提示RC/BTB2蛋白可能同时调节原纤毛和运动纤毛形成，该基因功能异常可能是导致纤毛病的重要原因。