3-D PROTEOME MAPPING OF BRAIN RELATED DISEASES

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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.