Genetic and alcohol regulation of brain RNA levels

大脑 RNA 水平的遗传和酒精调节

• 批准号: 6865676
• 负责人: SUSAN E. BERGESON
• 金额: $ 13.88万
• 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/6865676
• 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）： 该建议的目的是为Susan Bergeson博士提供职业发展 包括Adron Harris博士指导的机会，微阵列培训 与Vishy Iayer博士一起，并减少了教学工作，这将允许时间 制定独立研究计划。出色的研究 得克萨斯大学的环境，教职员工和设施将允许 扩展她在分子生物学方面的专业知识向新创新的 基因表达在酒精耐受性和依赖性中的作用分析。 提案的研究部分的目的是1）确定变化 在急性酒精暴露产生的脑mRNA水平中，到2） 鼠标行为遗传学与I的表达式变化链接到 适应分子神经记录，导致耐受性，依赖性和 致敏。这将通过mRNA差分显示（DD）完成 再加上DNA微阵列分析。已知定量性状的基因 位点（QTL）将获得优先级，并分析构造的异基小鼠 对于急性提取QTL，将提供概念验证。 QTL分析是 当前是映射包含基因的染色体区域的常见方法 在复合（多基因）哺乳动物特征中很重要，包括对 酒精，但只是理解基本机制的第一步 酒精的影响。几项表达研究，中殿鉴定了其基因 酒精的行动改变了mRNA水平，但没有清楚的了解 分子，酒精作用的后果已经出现。结合遗传 工具和分子技术为检测提供了逻辑策略 验证影响酒精相关的重要遗传差异 特质。 修改的DD分析，具有大多数筛查的能力 表达的成绩单以及识别插入和删除 多态性将在C57BL/6J（B6）和DBA/2J（D2）小鼠上完成，两只小鼠 QTL分析急性酒精产生的先天性菌株 提取。首先，B6，D2和先天性菌株的DD分析将 提供酒精反应和应变特定的表达差异。下一个， 微阵列将用于验证，剂量效果，时间表和 大脑区域特异性将通过添加DD隔离基因作为定义 DNA微阵列分析的其他目标为 通过聚类分析，阐明具有收敛调节的分子途径 由乙醇。另外，B6，D2之间的表达和序列多态性 这些先天性菌株对于众多具有或 将使用BXD策略。所有遗传和分析数据将被自由共享 通过MGI数据库（http：/www.informaticsjax.org）和Web中的沉积 在UT上创建的网站详细介绍了协议和结果；例如 请参阅：v.iyer，（http：//genome-www.stanford.edu/serum）。