Microarray Analysis of Alcohol Withdrawal Syndrome

戒酒综合症的微阵列分析

• 批准号: 6629702
• 负责人: SUSAN E. BERGESON
• 金额: $ 15万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2005-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6629702
• 关键词: alcoholism /alcohol abuse; behavioral /social science research tag; behavioral genetics; drug withdrawal; ethanol; genetic polymorphism; genetic screening; genetic susceptibility; immunocytochemistry; laboratory mouse; linkage mapping; microarray technology; neurochemistry; neurogenetics; northern blottings; pharmacogenetics; quantitative trait loci; substance abuse related behavior

DESCRIPTION: (provided by applicant) The goal of this proposal is twofold: 1. Identify gene expression changes specific to acute and/or chronic alcohol exposure and subsequent withdrawal in a discovery-based manner that will enhance the understanding of the molecular mechanisms of alcohol-related neuroadaptation. 2. Use DNA microarray technology as a genetic screening tool for Quantitative Trait Loci (QTL) analysis. A QTL is a site on a chromosome which statistically correlates with a trait under study and therefore likely contains a gene or genes which influence that trait. For disorders and complex traits that are inherited in a polygenic or non-Mendalian fashion (influenced by more than one gene or many genes), QTL analysis, recently successfully applied to mice, is currently a common method for mapping mammalian traits. However, once the QM have been mapped, determination of the genes or regulatory elements that underlie these regions of interest remains, for the most part, a formidable task. DNA microarray analysis applied to genetic mapping offers a logical strategy for the detection and validation of important genetic differences that influence a particular trait. The use of congenic strains of mice that have a chromosomal region of interest backcrossed onto a genetic background from a mouse that shows a different phenotype, and vice versa, allows direct comparison at the gene expression level, of that single chromosomal region. The primary objective is to profile genetic differences in brain regiospecific gene expression patterns that exist between C57BL/6J (B6) and DBA/2J (D2) mice (the two strains most commonly used in QTL mapping) as well as B6.D2 and D2.B6 congenic strains. B6 and D2 mouse strains have been extensively utilized in literally tens of thousands of genetic, behavioral, biochemical and pharmacological experiments by the scientific community. Analyzing polymorphisms in brain gene expression between these and several congenic strains will provide information of value to every study that has or will utilize a BXD strategy, including numerous alcohol-related studies. Another aim will be to focus the analysis directly on acute and chronic alcohol withdrawal by using unique selection lines of mice. In this case, microarray analysis will be used to identify genetic as well as alcohol-induced specific differences. Finally, Northerns, RPA, in situ hybridization, and where feasible Western blotting and immuno- histochemistry will be used to verify expression and consequent protein level changes relevant to the QTL mapping and alcohol effect DNA microarray-based analysis. All information on the genetic expression profiling of C57BL/6J and DBA/2J will be freely shared by deposition in the MGI data base (http:/www.informatics.jax.org) as well as the creation of a web site that details both the protocol and results; for an example see: V. Iyer, http://genome-stanford.edu/seruin/).

描述：(申请人提供) 这项提议的目标有两个：1.识别基因表达的变化 具体到急性和/或慢性酒精暴露和随后的戒断 一种基于发现的方式，将增强对分子的理解 酒精相关神经适应的机制。2.使用DNA微阵列 数量性状基因座(QTL)的遗传筛选技术 分析。QTL是染色体上的一个位置，它在统计上与 一种正在研究中的性状，因此很可能包含一个或多个基因 影响这一特点。对于疾病和复杂的特征来说， 多基因或非孟加拉风格(受多个基因或多个基因影响 基因)，QTL分析最近成功地应用于小鼠，目前是一种 哺乳动物性状图谱的常用方法。然而，一旦QM已经 绘制，确定构成这些基因或调控元件的基础 在很大程度上，感兴趣的地区仍然是一项艰巨的任务。脱氧核糖核酸 微阵列分析应用于遗传作图提供了一种合理的策略 检测和验证影响基因的重要遗传差异 独特的特质。利用具有染色体的同源小鼠品系 感兴趣区域从一只小鼠回交到遗传背景上 显示不同的表型，反之亦然，允许在 单个染色体区域的基因表达水平。初级阶段 目的是描述脑区特异性基因的遗传差异。 C57BL/6J(B6)和DBA/2J(D2)小鼠之间存在的表达模式( QTL定位中最常用的两个菌株)以及B6.D2和D2.B6 同源菌株。B6和D2小鼠品系已被广泛用于 从字面上看，成千上万的遗传、行为、生化和 科学界的药理实验。分析 这些基因与几个同源基因脑基因表达的多态性 菌株将为每一项已经或将要进行的研究提供有价值的信息 利用BXD策略，包括大量与酒精相关的研究。另一个 目标是将分析的重点直接放在急性和慢性酒精上 通过使用小鼠的独特选择系来戒断。在这种情况下，微阵列 分析将用于识别遗传和酒精诱导的特异性 不同之处。最后，Northerns、RPA、原位杂交以及在哪里 可行的Western blotting和免疫组织化学将用于验证 与QTL定位相关的表达和相应蛋白质水平的变化 基于DNA微阵列的酒精效应分析。上的所有信息 C57BL/6J和DBA/2J的基因表达谱将由 MGI数据库中的沉积物(http：//www.informatics.jax.org)以及 创建一个详细说明协议和结果的网站；对于 示例见：V.Iyer，http://genome-stanford.edu/seruin/).