Cell-type-specific and high-resolution profiling of postsynaptic proteomes altered in Alzheimer's disease model mice

项目总结/摘要 阿尔茨海默氏病（AD）是痴呆症的最常见原因，伴随着记忆和认知的进行性丧失。 功能协调发展的在几种AD模型小鼠中，记忆和认知功能的缺陷与AD的发生高度相关。 基础突触传递和突触可塑性受损。有趣的是，许多AD模型小鼠显示， 在b-淀粉样蛋白沉积发展之前，突触传递和可塑性的显著缺陷， AD病理学的典型标志。因此，突触功能障碍是AD的潜在重要组成部分 发病机制然而，为了在分子水平上了解AD中的突触功能障碍，我们面临着一个重大的挑战。 障碍，这是由于神经元异质性和突触历史（突触 可塑性）。因此，迫切需要开发一种生物化学方法，可以分析突触 分子成分的时空方式，以了解在物质上的突触分子的变化 参与AD。 这里，本申请的总体目标是开发系统型生物化学方法， 能够对突触后蛋白质组进行细胞类型特异性和高分辨率的分析。该项目的基本原理是 这些方法的发展很可能会促进分子均匀的突触后神经元的分离， 蛋白质组，这将最终提供更实质性的地图突触蛋白的变化，有助于 AD中的突触功能障碍。 为了实现总体目标，将追求以下两个具体目标： 使用Cre重组酶依赖性策略的突触后蛋白质组的分析方法;和2）开发高- 分辨率分析方法来识别突触下蛋白质组。突触后蛋白质组学的这一创新是 这将使我们能够从突触和突触下的特定群体中分离出突触后蛋白质组。 在AD模型小鼠中的结构域。AD模型小鼠中突触分子改变的更多实质性特征将 在分子水平上对AD中的突触功能障碍的理解提供了重大进展， 发现AD治疗靶点的机会。

PROJECT SUMMARY/ABSTRACT Alzheimer's disease (AD) is the most common cause of dementia with progressive loss of memory and cognitive functions. In several AD model mice, the deficits in memory and cognitive functions are highly correlated with an impairment in basal synaptic transmission and synaptic plasticity. Intriguingly, many of the AD model mice show significant deficits in synaptic transmission and plasticity before the development of b-amyloid deposits, which is a typical hallmark of AD pathology. Thus, synaptic dysfunction is a potentially important component of AD pathogenesis. However, to understand the synaptic dysfunction in AD at molecular levels, we face a significant obstacle, which is heterogeneity at multiple levels, due to neuronal heterogeneity and synaptic history (synaptic plasticity). There is, therefore, a critical need to develop a biochemical method that can analyze synaptic molecular constituents in a spatiotemporal manner to understand in substance the synaptic molecular alterations involved in AD. Here, the overall objective in this application is the development of systems-type biochemical methods that enable cell-type-specific and high-resolution profiling of postsynaptic proteomes. The rationale for this project is that these method developments are likely to advance the isolation of molecularly homogeneous postsynaptic proteomes, which will ultimately provide more substantive maps of synaptic protein changes that contribute to the synaptic dysfunction seen in AD. To attain the overall objectives, the following two specific aims will be pursued: 1) Develop cell-type-specific profiling methods of postsynaptic proteomes using Cre recombinase-dependent strategies; and 2) Develop high- resolution profiling methods to identify sub-synaptic proteomes. This innovation in postsynaptic proteomics is expected to allow us to isolate postsynaptic proteomes from the defined population of synapses and sub-synaptic domains in AD model mice. More substantive profiles of synaptic molecular alterations in AD model mice will provide a significant advance in the understanding of synaptic dysfunction in AD at molecular levels and new opportunities for discovering therapeutic targets of AD.