MOLECULAR BIOLOGY OF NICOTINIC RECEPTORS

烟碱受体的分子生物学

• 批准号: 6969112
• 负责人: SHERRY LEONARD
• 金额: $ 48.9万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-27 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6969112
• 关键词: RNA splicing; disease /disorder etiology; family genetics; gene expression; gene mutation; genetic polymorphism; genetic regulatory element; genetically modified animals; genotype; human subject; human tissue; laboratory mouse; messenger RNA; microarray technology; molecular biology; molecular pathology; nicotinic receptors; nucleic acid sequence; pathologic process; patient oriented research; phenotype; postmortem; receptor expression; schizophrenia

The Center proposes four postulates for investigating the role of genetic polymorphism in the alpha7 neuronal nicotinic acetylcholine receptor gene (CHRNA7) in schizophrenia: (1) identification and association of polymorphisms in CHRNA7 with schizophrenia, (2) demonstration of a functional effect of the polymorphism expression at the cellular level, (3) relation of the polymorphisms to deficits in brain function associated with the illness, and (4) reversal of the deficits in brain function by treatment directed at the cellular dysfunction caused by CHRNA7. This Project will address Postulates 1 and 2 by providing for the Center Investigators, genotyping of DNA from both live subjects and from postmortem brain, and in vitro evaluation of association with diagnosis and with function. CHRNA7 has been selected as the most likely candidate gene in the chromosome 15q14 linkage region by multiple biological and genetic studies. Functional mutations, consistent with reduced expression of the gene, were found in the proximal promoter region of the CHRNA7 gene. These are associated with both schizophrenia and the P50 deficit in the disease. Additional regulatory regions have been found in the 5' upstream region of the gene that contain polymorphic sites. This Project, Aim 1 will complete genotyping in 5' upstream regulatory regions, sequence the remaining sections of the gene in 10 probands from linked pedigrees, and investigate an associated intronic variant that may affect RNA splicing. SNPs in the four other genes in this region will be examined as well. As suggested by the Reviewers, we will also provide SNP genotyping for another gene with functional mutations, COMT, for association with schizophrenia and the phenotypes being investigated in this and other projects. For Postulate 2, Aim 2 will investigate the association of CHRNA7 mutations with CHRNA7 mRNA and protein expression in human postmortem brain of control and schizophrenic subjects. In Aim 3, we will further investigate the effects of CHRNA7 associated and functional mutations on the expression of other genes, utilizing microarray technology. The consequences of mutations in CHRNA7 on expression of CHRNA7, itself, and of other genes may have pathophysiological importance for the projects in the Center.

该中心提出了四个假设，用于调查精神分裂症中α 7神经元烟碱乙酰胆碱受体基因（CHRNA7）遗传多态性的作用：（1）CHRNA7多态性与精神分裂症的鉴定和关联，（2）在细胞水平上证明多态性表达的功能效应，（3）多态性与疾病相关的脑功能缺陷的关系，和（4）通过针对由CHRNA 7引起的细胞功能障碍的治疗逆转脑功能缺陷。本项目将通过为中心研究者提供活体受试者和死后大脑的DNA基因分型以及与诊断和功能相关性的体外评价来解决假设1和假设2。多项生物学和遗传学研究已将CHRNA7选为染色体15q14连锁区最有可能的候选基因。在CHRNA7基因的近端启动子区发现了与基因表达减少一致的功能突变。这些都与精神分裂症和疾病中的P50缺陷有关。在该基因的5'上游区域发现了含有多态性位点的另外的调控区。该项目，目标1将完成5'上游调控区的基因分型，对来自连锁家系的10个先证者的基因的其余部分进行测序，并研究可能影响RNA剪接的相关内含子变体。将检查该区域中其他四个基因中的SNP， 好.正如评审员所建议的那样，我们还将提供另一个具有功能突变的基因COMT的SNP基因分型，用于与精神分裂症以及本项目和其他项目中正在研究的表型相关。对于假设2，目标2将研究对照和精神分裂症受试者的人类死后脑中CHRNA7突变与CHRNA7 mRNA和蛋白质表达的关联。在目标3中，我们将利用微阵列技术进一步研究CHRNA7相关和功能突变对其他基因表达的影响。 CHRNA7突变对CHRNA7本身和其他基因表达的影响可能对该中心的项目具有病理生理学意义。