Creating atlases of gene expression using voxelation.

摘要

Gene expression signatures in the mammalian brain hold the key to understanding neural development and neurological diseases. The goal of this research is to provide atlases of gene and protein expression for the mouse genome in the brain. In order to accomplish this, voxelation was employed in combination with microarray and high-throughput mass spectrometry analysis. Voxelation involves dicing the brain into spatially registered voxels (cubes) at a resolution of 1 mm3. The studies presented here show gene and protein expression patterns in a coronal slice at the level of the striatum. At the most basic level, 20,000 gene and 1,500 protein expression images for this coronal slice are now available to the neuroscience community for reference to other studies. Good reliability of the microarray results were confirmed using multiple replicates, subsequent quantitative RT-PCR voxelation, mass spectrometry voxelation and publicly available in situ hybridization data. Higher levels of analysis identify clusters of genes that have unexpected patterns, such as a dorsal/ventral gradient of expression not restricted to known anatomical boundaries. Additionally, this cross-modality study investigates the relationship between transcript and protein levels which have previously been thought to be difficult to relate directly.;The final study of this dissertation involves characterizing two pharmacological mouse models of Parkinson's disease using transcriptomic and proteomic techniques. Microarray and high-throughput mass spectrometry analyses of the mouse striatum demonstrate a large number of genes and proteins that are differentially regulated in response to treatment. Again, this cross-modality study provides insights into potential translation controls of drug response. MicroRNA suppression of protein expression is a strong candidate as a controlling agent in this process. The findings of this study provide the neuroscience community with further insights into the molecular architecture of the brain in two pharmacological models of Parkinson's disease in the mouse.