Generation and analysis of candidate genes from substantia nigra

该项目的目的是研究黑质中的差异基因表达，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系统疾病。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，在患病个体中，该组织中表达显著上调或下调的基因构成了PD及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）是一项强大的技术，通过克隆和测序与每个基因相关的大量独特的13碱基对标签，可以定量测定给定组织中存在的所有转录本。Vance博士和他的合作者计划使用SAGE来比较从三种不同来源分离的黑质中表达的基因群体：正常对照个体、路易体阳性PD患者和FTDP患者（一种路易体阴性神经退行性疾病，也以黑质胶质瘤为特征）。最后一组样本将允许研究人员控制PD黑质中神经元代表性不足的混淆效应。在疾病状态下，黑质中表达过少或过少的基因将被视为潜在的候选基因，正常黑质中特异性表达的任何基因也将被视为潜在的候选基因。这些候选基因将在项目I中使用项目I中概述的特发性PD患者的亚基关联分析来测试与PD的关联。这种双筛选-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.