活细胞高分辨率成像解析clathrin介导的内吞囊泡形成早期内体的分子机制

Clathrin介导的内吞是最重要的内吞途径之一，在组织器官发育、细胞信号转导、神经传导等过程中具有重要作用。早期内体是内吞后物质在细胞内分选的第一站，近些年的研究和我们的前期研究均提示在经典的早期内体之外，还可能存在定位分布、动力学、结构和功能等不同的其他类型的早期内体结构。然而，clathrin内吞囊泡是如何形成早期内体、特别是不同类型早期内体的机制仍然未知。针对这一重要科学问题，本项目中我们将整合基因编辑、荧光探针设计和活细胞超高分辨率三维成像等多种技术，全面解析各种早期内体在细胞内精确定位、clathrin内吞囊泡形成早期内体的动态过程和分子调控机制。因此，本项目是利用多学科交叉的研究手段研究一个重要生物过程在一个核心细胞器形成中的动态机制，具有很强的创新性和重要科学意义。研究结果将揭示clathrin内吞囊泡形成早期内体的分子机制，也将为内吞膜转运领域提供新的研究策略和技术方法。

Clathrin-mediated endocytosis is one of the most important endocytic pathway, which regulates tissue and organ development, cell signaling, neural transduction and many other essential cellular processes. Early endosomes are the first cell sorting station after cargo uptake by endocytosis. Many recent researches including ours have suggested that early endosomes are highly dynamic and have several different types with different protein markers, dynamics, locations, subcellular structures and functions. However, exactly how the clathrin-mediated endocytic vesicles form early endosomes especially different types of early endosomes is largely unknown. In this project, we will employ a series of cutting-edge techniques such as genome editing, sensor design, and live-cell high-resolution quantitative imaging to perform a compressive study on the precise intracellular localizations of different types of early endosomes, and the dynamic processes and molecular mechanisms of early endosome formation from clathrin-mediated endocytic vesicles. Thus, by using these cutting-edge multidisciplinary approaches, this project will reveal the molecular mechanisms of a crucial biological process in the generation of a critical organelle. It will uncover novel molecular basis of early endosome formation from clathrin-mediated endocytic vesicles and will provide new tools and strategies for the membrane trafficking research field.