3-D PROTEOME MAPPING OF BRAIN RELATED DISEASES

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。加州大学洛杉矶分校和PNNL的研究人员正在合作开发第一个小鼠大脑转录组和蛋白质组结构的集成三维(3D)模型，以揭示正常大脑中mRNA和蛋白质的表达模式，并将正常区域的表达模式与明显受到疾病、药物滥用或其他创伤损害的相似区域进行比较。一种双重策略被用来：1)系统地绘制1立方毫米大小的空间注册体积元素(称为体素)中的蛋白质和信使核糖核酸的表达水平，以及2)研究已知受特定疾病影响的大脑区域(例如，纹状体)中信使核糖核酸和蛋白质的调节。我们最初的初步研究将针对甲基苯丙胺和1-甲基-4-苯基-1，2，3，6-四氢吡啶(MPTP)引起的脑损伤，这种损伤产生类似帕金森病(PD)的生理影响，主要损害大脑纹状体。其目标是识别和量化受损纹状体中蛋白质的不同丰度，并建立与帕金森病类型症状最密切相关的蛋白质清单。为了定量绘制体素内的蛋白质表达水平图(包含少于20微克的蛋白质)，我们正在开发新的自动化微型样本处理平台，以高效处理从单个小鼠脑中获得的~600个体素，改进我们的高分辨率色谱分离以实现高通量分析，并优化LC-FTICR以实现最高的灵敏度，以提供整体高覆盖的蛋白质组。

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 (containing less than 20 micrograms of proteins), we are developing new automated microscale sample-handling platform for efficient processing the ~600 voxels obtained from a single mouse brain, refining our high-resolution chromatographic separations for high throughput analyses, and optimizing the LC-FTICR for maximum sensitivity to provide overall high coverage of the proteome.