Genetic and alcohol regulation of brain RNA levels

大脑 RNA 水平的遗传和酒精调节

• 批准号: 6620451
• 负责人: SUSAN E. BERGESON
• 金额: $ 13.39万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-03-01 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6620451
• 关键词: alcoholism /alcohol abuse; behavior test; behavioral genetics; behavioral habituation /sensitization; brain; differential display technique; dosage; drug tolerance; ethanol; gene expression; genetic polymorphism; genetic regulation; genetic strain; immunologic techniques; laboratory mouse; linkage mapping; messenger RNA; microarray technology; molecular biology; molecular biology information system; neuroanatomy; neurotoxicology; nucleic acid hybridization; nucleic acid sequence; postdoctoral investigator

DESCRIPTION (provided by applicant): The goal of this proposal is to provide Dr. Susan Bergeson career development opportunities including mentoring by Dr. Adron Harris, microarray training with Dr. Vishy Iayer, and a reduction in teaching effort that will allow time to develop an independent research program. The excellent research environment, faculty and facilities of the University of Texas will allow expansion of her expertise in molecular biology toward a new and innovative analysis of the role of gene expression in alcohol tolerance and dependence. The aims of the research component of the proposal are to 1) Identify changes in brain MRNA levels produced by acute alcohol exposure and to 2) Utilize mouse behavioral genetics to link I coordinated changes in expression to adaptations in molecular neurocircuitry that cause tolerance, dependence and sensitization. This will be accomplished by MRNA differential display (DD) coupled with DNA microarray analysis. Genes within known Quantitative Trait Loci (QTL) will be given priority and analysis of congenic mice constructed for acute withdrawal QTLs will provide a proof-of-concept. QTL analysis is currently a common method for mapping chromosomal regions that contain genes important in complex (polygenic) mammalian traits, including responses to alcohol but is only a first step in understanding mechanisms underlying alcohol's effects. Several expression studies, nave identified genes whose mRNA levels are changed by alcohol's actions yet no clear picture of the molecular, consequences of alcohol action have emerged. Combining genetic tools and molecular techniques offers a logical strategy for the detection and validation of important genetic differences that influence alcohol-related traits. A modified DD analysis, which has the capacity to screen most expressed transcripts as well as identify insertion and deletion polymorphisms, will be completed on C57Bl/6J (B6) and DBA/2J (D2) mice and two congenic strains created as a result of QTL analysis for acute alcohol withdrawal. First, DD analysis of B6, D2, and the congenic strains will provide alcohol-responsive and strain specific expression differences. Next, microarrays will be used for validation, and dose-effect, time-course, and brain region specificity will be defined by adding the DD isolated genes as additional targets for DNA microarrays analysis providing sufficient power to elucidate, by cluster analyses, molecular pathways with convergent regulation by ethanol. In addition, expression and sequence polymorphisms between B6, D2 and these congenic strains will be of value to numerous studies that have or will use a BXD strategy. All genetic and profiling data will be freely shared by deposition in the MGI data base (http:/www.informaticsjax.org) and a web site created at UT that details both the protocol and results; for an example see: V.Iyer,(http://genome-www.stanford.edu/serum).

描述（由申请人提供）： 该提案的目标是为 Susan Bergeson 博士提供职业发展 机会包括 Adron Harris 博士的指导、微阵列培训 与 Vishy Iayer 博士合作，减少教学工作量，留出时间 制定独立的研究计划。优秀的研究 德克萨斯大学的环境、师资和设施将允许 将她在分子生物学方面的专业知识扩展到新的和创新的领域 分析基因表达在酒精耐受和依赖中的作用。 该提案研究部分的目标是 1) 识别变化 急性酒精暴露产生的大脑 mRNA 水平以及 2) 利用 小鼠行为遗传学将 I 协调表达的变化与 分子神经回路的适应导致耐受性、依赖性和 敏化。这将通过 mRNA 差异显示 (DD) 来完成 结合 DNA 微阵列分析。已知数量性状中的基因 优先考虑位点（QTL）并分析构建的同系小鼠 对于急性戒断 QTL 将提供概念验证。 QTL分析是 目前绘制包含基因的染色体区域的常用方法 对复杂（多基因）哺乳动物特征很重要，包括对 酒精，但这只是理解潜在机制的第一步 酒精的影响。一些表达研究，nave 确定了基因 mRNA 水平会因酒精的作用而改变，但尚不清楚具体的情况 酒精作用的分子后果已经显现。结合遗传 工具和分子技术为检测和分析提供了逻辑策略 验证影响酒精相关的重要遗传差异 特征。 改进的 DD 分析，能够筛选大多数 表达的转录本以及识别插入和删除 多态性，将在 C57Bl/6J (B6) 和 DBA/2J (D2) 小鼠和两只 急性酒精 QTL 分析产生的同源菌株 撤回。首先，对B6、D2和同系株进行DD分析 提供酒精响应和菌株特异性表达差异。下一个， 微阵列将用于验证、剂量效应、时程和 大脑区域特异性将通过添加 DD 分离基因来定义： DNA 微阵列分析的其他目标提供了足够的能力 通过聚类分析阐明具有收敛调控的分子途径 通过乙醇。此外，B6、D2之间的表达和序列多态性 这些同源菌株对于许多具有或 将使用 BXD 策略。所有遗传和分析数据将免费共享 通过沉积在 MGI 数据库 (http://www.informaticsjax.org) 和网络中 UT 创建的网站详细介绍了协议和结果；举个例子 参见：V.Iyer，(http://genome-www.stanford.edu/serum)。