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：（由申请人提供） 该提案的目标有两个： 1. 识别基因表达变化 特定于急性和/或慢性酒精暴露以及随后的戒断 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）的遗传筛选工具 分析。 QTL 是染色体上的一个位点，与 正在研究的性状，因此可能包含一个或多个基因 影响该特质。对于遗传性疾病和复杂特征 多基因或非孟德尔时尚（受多个基因或多个基因影响） 基因），QTL 分析最近成功应用于小鼠，目前是一种 绘制哺乳动物性状的常用方法。然而，一旦 QM 完成 绘制、确定这些基因或调控元件的基础 在很大程度上，感兴趣的区域仍然是一项艰巨的任务。脱氧核糖核酸 应用于遗传图谱的微阵列分析为以下方面提供了逻辑策略： 影响基因的重要遗传差异的检测和验证 特定的特质。使用具有染色体的同系小鼠品系 感兴趣的区域与小鼠的遗传背景回交， 显示不同的表型，反之亦然，允许直接比较 该单个染色体区域的基因表达水平。初级 目的是分析大脑区域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 小鼠品系已广泛用于 实际上有数以万计的遗传、行为、生化和 科学界的药理实验。分析中 这些和几个同类之间的大脑基因表达多态性 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 利用 BXD 策略，包括大量与酒精相关的研究。其他 目的是将分析直接集中在急性和慢性酒精上 通过使用独特的小鼠选择系进行戒断。在这种情况下，微阵列 分析将用于识别遗传以及酒精引起的特定 差异。最后，北方人，RPA，原位杂交，以及哪里 可行的蛋白质印迹和免疫组织化学将用于验证 与 QTL 作图相关的表达和随后的蛋白质水平变化 以及基于 DNA 微阵列的酒精效应分析。有关的所有信息 C57BL/6J 和 DBA/2J 的基因表达谱将免费共享 沉积在 MGI 数据库 (http://www.informatics.jax.org) 以及 创建一个详细介绍协议和结果的网站；对于一个 示例参见：V. Iyer，http://genome-stanford.edu/seruin/）。