Microarray Analysis of Alcohol Withdrawal Syndrome

戒酒综合症的微阵列分析

• 批准号: 6334233
• 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 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6334233
• 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） 该提案的目标有两个：1。识别基因表达变化 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， 酒精相关的神经适应机制2.使用DNA微阵列 技术作为数量性状基因座（QTL）遗传筛选工具 分析. QTL是染色体上的一个位点，其统计学上与 研究中的性状，因此可能包含一个或多个基因， 影响这种特质。对于遗传的疾病和复杂特征， 多基因或非Mendalian时尚（受一个以上的基因或许多基因的影响） 基因），QTL分析，最近成功地应用于小鼠，目前是一个 绘制哺乳动物特征的常用方法。然而，一旦QM 映射，确定这些基因或调控元件的基础 在大多数情况下，感兴趣的区域仍然是一项艰巨的任务。DNA 微阵列分析应用于遗传作图提供了一种逻辑策略， 检测和验证重要的遗传差异，影响 特别的特质使用具有染色体畸变的小鼠的同类品系， 将感兴趣的区域回交到来自小鼠的遗传背景上， 显示了不同的表型，反之亦然，允许直接比较 基因表达水平。主 目的是分析脑区域特异性基因的遗传差异， C57 BL/6 J（B6）和DBA/2 J（D2）小鼠之间存在的表达模式（ QTL作图中最常用的两个菌株）以及B6.D2和D2.B6 同类株系B6和D2小鼠品系已被广泛用于 数以万计的基因，行为，生化和 药理学实验，科学界。分析 这些基因与几个同类基因之间的脑基因表达多态性 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。 使用BXD策略，包括许多与酒精相关的研究。另一 目标将是直接集中分析急性和慢性酒精中毒 通过使用小鼠的独特选择系来撤回。在这种情况下，微阵列 分析将用于确定遗传以及酒精诱导的特异性 差异最后，Northerns，RPA，原位杂交，以及 将采用可行的Western印迹和免疫组织化学方法进行验证 表达和随后与QTL定位相关的蛋白质水平变化 和酒精效应DNA微阵列分析。上的所有信息 C57 BL/6 J和DBA/2 J的基因表达谱将由以下人员免费共享： 沉积在MGI数据库（http：/www.informatics.jax.org）以及 建立一个网站，详细说明协议和结果;对于一个 实例参见：V. Iyer，http：//genome-stanford.edu/seruin/）。