血影蛋白αⅡ与肌动蛋白作用维持静纤毛稳定

项目名称： 血影蛋白αⅡ与肌动蛋白作用维持静纤毛稳定

项目编号： No.81470703

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 于栋祯

作者单位： 上海交通大学

项目金额： 73万元

中文摘要： 肌球蛋白VI基因突变导致毛细胞静纤毛异常和感音神经性聋，静纤毛是以β和γ肌动蛋白为骨架。本课题组前期进行了内耳毛细胞肌球蛋白VI相互作用蛋白的研究，通过免疫共沉淀、质谱分析、Western blot、免疫荧光技术等手段，发现血影蛋白αII与肌球蛋白VI作用，血影蛋白αⅡ与肌球蛋白VI和肌动蛋白在表皮板共表达，且血影蛋白αⅡ在静纤毛根部强表达，提示血影蛋白αⅡ参与维持静纤毛结构的稳定。然而血影蛋白αⅡ在静纤毛结构的维持和听功能中的作用尚不清楚。本课题拟研究血影蛋白αⅡ下调对体外培养基底膜肌动蛋白的表达和毛细胞静纤毛的影响，并拟利用突变小鼠和斑马鱼基因敲除模型研究血影蛋白αⅡ下调对肌动蛋白的表达以及毛细胞形态包括静纤毛形态和听功能的影响。本课题旨在揭示血影蛋白αⅡ在内耳毛细胞静纤毛稳定中的作用，为防治静纤毛异常导致的耳聋提供理论基础。

中文关键词： 毛细胞；分子机制；血影蛋白αⅡ；静纤毛；小鼠模型

英文摘要： In previous study, we identified a Myosin VI interactive protein, spectrinαII. Then we studied the cellular and subcellular distributions of spectrin αⅡ by immunofluorescence analysis using confocal microscopy and found spectrin αⅡ is strongly expressed in the region of cuticular plate , and especially around the rootlets of stereocilia of inner and outer hair cells, suggesting that spectrin αⅡ plays an important role in maintaining the structure of stereocilia. However, it needs to further confirm this role and it is unclear whether mutation in spectrin αⅡ can lead to hearing loss. In this project, we are going to down regulate spectrin αⅡ gene by talen technique in vitro cultured basilar membranes, and observe the morphological changes of hair bundles of inner and outer hair cells. Then, we will observe morphological changes and auditory funtion in spectrin αⅡ mutant mouse. At last, we will down regulate spectrin αⅡ in Zebrafish and observe the development changes of neuromast and inner ear, otolith formation and hair bundles. This project is aimed to identify the action between spectrinαⅡ and actin, and explore the role of sepctrinαⅡ in maintaining the stability of hair cells apical structure. The implementation of this project helps to uncover the role of spectrinαⅡ in stereocilia stability, which will provide cues for the prevention and treatment of stereocilia dysfuntion related sensorineural hearing loss.

英文关键词： hair cells;molecular mechnism;spectrin αⅡ;stereocilia;mouse model