Tks4参与Reelin信号通路调节端脑投射神经元发育的研究

Frank-Ter Harr 综合征是人类常染色体隐性遗传病，患者表现出系列发育性异常，包括脑发育异常。该疾病致病因子所编码蛋白为TKS4，具有一个PX结构域以及4个SH3结构域，主要通过调节胞内蛋白复合物的膜分布来参与细胞内信号转导过程。单细胞测序结果显示Tks4主要在迁移神经元中表达，同时在神经元中与Crk/CrkL结合形成复合物，因此可能参与Reelin信号通路来调节神经元迁移。本项目拟在细胞水平上分析Tks4调节Crk/CrkL膜分布的具体方式；构建基因敲除小鼠分析端脑板层结构；另外联合应用FlashTag以及脑片透明化技术，建立精准分析神经元迁移的新技术手段，全面分析Tks4缺失对于神经元迁移的影响。最终通过小鼠行为变化解析Tks4调节脑发育以及脑功能的重要作用。本项目的研究结果有望进一步完善Reelin信号通路的调控方式，同时为该疾病的诊断和治疗提供重要理论基础。

Frank-Ter Harr syndrome is an autosomal recessive genetic disease in human. The patient shows a series of developmental abnormalities, including brain dysplasia. The disease pathogenic factor encodes protein-Tks4, which has one PX domain and four SH3 domains. It mainly plays roles in the signaling transduction process by regulating the membrane distribution of intracellular protein complexes. Single cell sequencing data showed that Tks4 highly expressed in the migrating neurons during brain development, and Tks4 binds with Crk/CrkL to form a protein complex in neurons. Therefore, Tks4 may be involved in the Reelin signaling transduction pathway to regulate neuronal migration. In this project, we plan to analyze the regulatory role of Tks4 in mediating Crk/CrkL membrane distribution at the cell level; construct Tks4 gene knockout/ conditional knockout mice to analyze the lamellar structure of telencephaly; in addition, we propose to apply FlashTag and ultrafast optical clearing method (FOCM) technology to establish a new strategy for precise analysis of neuronal migration, and comprehensively analyze the functions of Tks4 on neuronal migration. Finally, the important roles of Tks4 in brain development and brain function will be analyzed via analyses of mouse behavioral tests. The results of this project are expected to further delineate the fine regulation of Reelin signaling pathway, and provide an important theoretical basis for the diagnosis and treatment of Frank-Ter Harr syndrome.