Generation and analysis of candidate genes from substantia nigra

这个项目的目标是研究黑质差异基因的表达，以产生帕金森病(PD)的候选基因。帕金森病是一种以黑质病理性变性为特征的遗传性神经疾病。研究人员假设，变性与该组织中基因表达模式的改变有关；因此，在受影响的个体中，该组织中表达显著上调或下调的基因构成了帕金森病和相关疾病的良好候选基因。其他候选基因将被确定为那些在正常黑质中表达水平高于正常人脑其他区域的基因。基因表达序列分析(SAGE)是一种强大的技术，它通过克隆和测序与每个基因相关的大量唯一的13个碱基对标签来定量确定特定组织中存在的所有转录本。万斯博士和他的合作者计划使用SAGE来比较来自三个不同来源的黑质中表达的基因群体：正常对照组、路易体阳性帕金森病患者和FTDP患者。FTDP是一种路易体阴性神经退行性疾病，也是一种以黑质胶质增生为特征的疾病。这最后一组样本将使研究人员能够控制PD黑质神经元表达不足的混杂效应。在疾病状态下在黑质中表达不足或过度的基因将被认为是潜在的候选基因，任何在正常黑质中特异表达的基因都将被认为是潜在的候选基因。这些候选基因将在项目I中使用基于亚基的关联分析在特发性帕金森病患者中进行测试，如项目I中概述的那样。这种双重筛选-在适当组织中的差异表达，以及与疾病的统计关联-将是选择候选基因的非常强大的机制。

The goal of this project is to investigate differential gene expression in the substantia nigra in order to generate candidate genes for Parkinson's Disease (PD), a heritable neurological disorder characterized by pathological degeneration of substantia nigra. The investigators hypothesize that degeneration is associated with an altered pattern of gene expression in this tissue; therefore, genes whose expression in this tissue is significantly up-or down-regulated in affected individuals constitute good candidate genes for PD and related disorders. Additional candidate genes will be identified as those genes that are expressed in the normal substantia nigra at a higher level than in other regions of normal human brains. Serial Analysis of Gene Expression (SAGE) is a powerful technique that allows quantitative determination of all transcripts present in a given tissue by cloning and sequencing large numbers of unique 13-base pair tags associated with each gene. Dr. Vance and his collaborators plan to use SAGE to compare the population of genes expressed in substantia nigra isolated from three different sources: normal control individuals, individuals affected with Lewy body positive PD, and individuals affected with FTDP, a Lewy body negative neurodegenerative disease also characterized by gliosis of the substantia nigra. This last group of samples will allow the investigators to control for the confounding effects of under-representation of neurons in the PD substantia nigra. Genes that are under- or over-expressed in the substantia nigra in the disease-state will be considered potential candidate genes, as will any genes expressed specifically in the normal substantia nigra. These candidate genes will be tested for association with PD in Project I using sub-based association analysis among patients with idiopathic PD as outlined in Project I. This double screen-differential expression in the appropriate tissues, as well as statistical association with disease-will be a very powerful mechanism for selecting candidate genes.