Mints: Adaptor Proteins Coupling APP of Alzheimer's Disease to the Synapse

描述（申请人提供）：大脑功能需要神经元之间的适当网络和通信。神经功能发育和维持不当导致神经系统异常。因此，大脑中生理到病理过程的潜在机制尚不清楚。目前，Thomas c.s¿dhof博士的实验室使我能够将突触传递的基本事件扩展到阿尔茨海默病的临床相关问题。我们已经确定了一个名为Mints的重要衔接蛋白家族，它与突触功能的耦合有关，如蛋白质靶向神经末梢和神经传递，以及与阿尔茨海默病相关的淀粉样前体蛋白（APP）加工的调节。为了直接确定薄荷的功能，我们产生了缺乏单个薄荷蛋白（同种异构体1-3）或所有可能的薄荷家族成员组合的小鼠。我们现在可以直接研究：(1)通过细胞膜表面生物素化来靶向膜蛋白，并从形态学上观察薄荷相互作用蛋白的表达和细胞分布；(2)利用海马体切片电生理记录和培养神经元的光学记录技术观察突触囊泡的动力学，从功能上检查突触传递。我们将通过电子显微镜和E-PTA染色来表征突触连接的结构动态，并量化突触的形态参数。为了探索薄荷和APP加工的意义，我们培育了薄荷缺乏的小鼠，这些小鼠携带了一种共表达突变APP和早老素1的转基因。我们将结合形态学和生化技术，通过检测年龄依赖性APP蛋白水解和β淀粉样蛋白沉积来研究导致疾病状态的致病事件。这些研究不仅将阐明Mints在靶向和突触传递中的作用，而且将拓宽我们对Mints和Mints生物学的认识。APP在阿尔茨海默病中的作用。我的长期目标是整合我在过去的研究生和博士后培训中所学到的工具和概念方法，追求我对神经元可塑性和神经退行性疾病的分子机制的理解。这个奖项将让我有一个过渡期，在此期间我可以扩展我的知识和技术基础，成为一名独立的首席研究员。

DESCRIPTION (provided by applicant): Brain function requires the proper networking and communication between neurons. Improper development and maintenance of neuronal function leads to neurological abnormalities. As such, mechanisms underlying physiological to pathological processes in the brain are not clear. Currently, the laboratory of Dr. Thomas C. S¿dhof has allowed me to expand my knowledge in basic events of synaptic transmission to clinically associated problems in Alzheimer's disease. We have identified an essential family of adaptor proteins named Mints that have been implicated in coupling synaptic functions such as targeting of proteins to nerve terminals, and neurotransmission, to the regulation of amyloid precursor protein (APP) processing relating to Alzheimer's disease. To ascertain Mints function directly, we have generated mice lacking individual Mint proteins (isoforms 1-3), or all possible combination of Mint family members. We can now directly study: (1) membrane protein targeting by surface biotinylation of cell membranes, and morphologically examine the expression and cellular distribution of proteins that Mints interact with; (2) functionally examine synaptic transmission by using hippocampal slice electrophysiological recordings, and optical recording techniques of cultured neurons to look at kinetics of synaptic vesicles. We will characterize the structural dynamic of synaptic junctions by electron microscopy, and E-PTA staining to quantify morphological parameter of synapses. To explore the significance of Mint and APP processing, we have generated mice deficient of Mints which carry a transgene that coexpresses mutant APP, and presenilin 1. We will study the pathogenic events leading to disease state by examining age-dependent APP proteolysis and amyloid beta deposition by combining morphological and biochemical techniques. These studies will not only clarify the function of Mints in targeting, and synaptic transmission, but will broaden our understanding in the biology of Mints and. APP in Alzheimer's disease. My long-term goal is to pursue my understanding of molecular mechanisms underlying neuronal plasticity, and neurodegenerative diseases integrating the tools and conceptual approaches that I have learned and gained over my past graduate and postdoctoral training. This award will allow me to have a transition period during which I can expand my knowledge and technical foundations to become an independent principal investigator.