Genetic and Proteomic Analysis of Protein Sorting and Retrieval During Synaptic Vesicle Endocytosis

神经系统的一个独特特征是大量的细胞-细胞连接，称为突触。 在突触处，神经细胞彼此之间以及与其他靶细胞进行通信。 在化学突触中，当激活时，神经细胞从小突触囊泡（SV）中释放递质，然后回收这些囊泡以重新使用。 因此，SV再循环在神经元功能中起主要作用。 网格蛋白及其相关分子是囊泡循环的关键参与者。 然而，SV回收的分子机制尚未完全了解。 该项目旨在了解SV蛋白如何被招募并回收到新形成的囊泡中。 来自张博士实验室和其他人的遗传和生物化学研究表明，网格蛋白辅助蛋白AP 180调节网格蛋白介导的SV再循环的速率和保真度。 这个项目将测试的假设，即AP 180也发挥了作用，帮助回收囊泡蛋白在囊泡回收。 他们将研究在缺乏AP 180的突变果蝇（果蝇）中SV的组成是否以及如何改变，并将研究AP 180可能与囊泡蛋白相互作用的模式。 这些研究有望促进对SV再循环和神经细胞功能的理解。 该项目还将研究与教育、培训和对当地社区的宣传紧密结合起来。 他培训和指导过本科生、研究生和博士后研究员，包括来自代表性不足群体的妇女和学生。 11名本科生与他共同撰写了之前NSF赞助的项目，其中一人是第一作者。 在此资助期间，他将继续在学生辅导和课程开发方面的教育工作。 此外，他还将接触当地学校，并通过一个由萨姆诺布尔俄克拉荷马州自然历史博物馆组织的暑期项目培训小学科学教师。

A unique feature of the nervous system is the vast number of cell-cell connections called synapses. At synapses, nerve cells communicate with each other and with other target cells. At chemical synapses, and upon activation, nerve cells release transmitter from small synaptic vesicles (SVs) and then recycle these vesicles for reuse. Hence, SV recycling plays a major role in neuronal function. Clathrin and its associated molecules are critical players in vesicle recycling. However, the molecular mechanisms by which SVs are recycled are not fully understood. This project is directed towards understanding how SV proteins are recruited and recycled into newly formed vesicles. Genetic and biochemical studies from Dr. Zhang's laboratory and others suggest that the clathrin-accessory protein AP180 regulates both the rate and the fidelity of clathrin-mediated recycling of SVs. This project will test the hypothesis that AP180 also plays a role in helping retrieve vesicle proteins during vesicle recycling. They will be examining whether and how the composition of SVs is altered in mutant fruit flies (Drosophila) deficient of AP180 and also will investigate the mode by which AP180 might interact with vesicular proteins. These studies are expected to advance the understanding of SV recycling and nerve cell function. This project also closely integrates research with education, training, and outreach to local communities. He has trained and mentored undergraduates, graduates, and postdoctoral fellows, including women and students from underrepresented groups. Eleven undergraduates were co-authors with him on a prior NSF-sponsored project, including one as first author. During this funding period, he will continue his educational efforts in student mentoring and course development. In addition, he will be reach out to local schools and train elementary science teachers through a summer program organized by the Sam Noble Oklahoma Museum of Natural History at OU.