3D MOUSE BRAIN PROTEOME MAPPING OF NORMAL AND PARKINSON'S DISEASE MOUSE MODELS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 来自加州大学洛杉矶分校和PNNL的研究人员正在合作开发小鼠大脑转录组和蛋白质组结构的第一个集成三维（3-D）模型，以揭示整个正常大脑中mRNA和蛋白质的表达模式，并将正常区域的表达模式与疾病，药物滥用或其他创伤明显受损的类似区域进行比较。 采用双重策略：1）系统地绘制具有1立方毫米大小的空间配准体积元素（称为体素）中的蛋白质和mRNA表达水平，以及2）研究脑区域（例如，纹状体）已知特别受某些疾病的影响。 我们最初的试点研究将是甲基苯丙胺和1-甲基-4-苯基-1，2，3，6-四氢吡啶（MPTP）诱导的脑损伤，产生类似于帕金森病（PD）的生理影响，主要损害大脑的纹状体。 目的是确定和量化受损纹状体内蛋白质的差异丰度，并建立与PD型症状最密切相关的蛋白质列表。 为了定量绘制体素内的蛋白质表达水平，我们正在开发新的自动化微尺度样品处理和高通量分析平台，用于有效处理和分析从单个小鼠大脑获得的约600个体素，以生成蛋白质分布的3-D图。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Researchers from UCLA and PNNL are collaborating to develop the first integrated three-dimensional (3-D) model of the transcriptome and proteome constructs of the mouse brain in order to reveal the expression patterns of mRNA and proteins throughout the normal brain and to compare the expression patterns of normal regions to similar regions apparently damaged from disease, drug abuse, or other trauma. A two-fold strategy is being employed to: 1) systematically map the protein and mRNA expression levels in spatially registered volume elements (known as voxels) with 1 cubic millimeter size, and 2) investigate the regulation of mRNA and proteins in brain regions (e.g., striatum) known to be specifically affected by certain diseases. Our initial pilot study will be on methamphetamine and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced brain damage, which produces physiological affects similar to Parkinson's Disease (PD) and primarily damages the striatum of the brain. The goal is to identify and quantify the differential abundance of proteins within the damaged striatum and establish a list of proteins that are most intimately associated with PD-type symptoms. In order to quantitatively map protein expression levels within voxels, we are developing new automated microscale sample-processing and high throughput analysis platform for efficient processing and analyzing the ~600 voxels obtained from a single mouse brain to generate 3-D maps of protein distributions.