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适当的网络和通信。神经功能发育和维持不当会导致神经功能异常。因此，大脑中生理和病理过程的潜在机制尚不清楚。目前，托马斯·C·S博士的实验室使我能够将我在突触传递基本事件方面的知识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的重要适配器蛋白家族，它们参与了突触功能的偶联，如将蛋白质靶向神经末梢和神经传递，以调节与阿尔茨海默病相关的淀粉样前体蛋白(APP)的处理。为了直接确定薄荷的功能，我们培育了缺乏单个Mint蛋白(亚型1-3)或所有可能的Mint家族成员组合的小鼠。我们现在可以直接研究：(1)通过细胞膜表面的生物素化来靶向膜蛋白，并从形态上检测Mints与之相互作用的蛋白的表达和细胞分布；(2)利用海马片电生理记录和光学记录技术从功能上研究突触传递，观察突触小泡的动力学。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，以量化突触的形态参数。为了探索薄荷和APP加工的意义，我们产生了薄荷缺失的小鼠，这些小鼠携带着共同表达突变APP和早老素1的转基因。我们将结合形态学和生化技术，通过检测依赖年龄的APP蛋白分解和淀粉样β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薄荷糖在靶向和突触传递中的功能，而且将拓宽我们对薄荷糖和薄荷糖生物学的理解。阿尔茨海默病中的APP。我的长期目标是追求我对神经元可塑性和神经退行性疾病潜在的分子机制的理解，整合我在过去的研究生和博士后培训中学到的工具和概念方法。这一奖项将使我有一个过渡期，在此期间我可以扩大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.