Genetic and alcohol regulation of brain RNA levels

描述（由申请人提供）： 本提案的目标是为苏珊·伯格森博士提供职业发展机会 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， 开发一个独立的研究项目优秀的研究 德克萨斯大学的环境，教师和设施将允许 她在分子生物学的专业知识扩展到一个新的和创新的 分析基因表达在酒精耐受和依赖中的作用。 该提案的研究部分的目的是：1）确定变化 在大脑mRNA水平产生的急性酒精暴露和2）利用 小鼠行为遗传学将I协调的表达变化与 分子神经回路的适应性，导致耐受性，依赖性和 致敏这将通过mRNA差异显示（DD）来实现。 再加上DNA微阵列分析。已知数量性状中的基因 基因座（QTL）将优先考虑和同类小鼠的分析构建 急性戒断的QTL将提供概念验证。QTL分析是 目前，一种用于绘制包含基因的染色体区域的常用方法 重要的复杂（多基因）哺乳动物性状，包括反应， 酒精，但这只是了解潜在机制的第一步。 酒精的影响。几项表达研究，初步鉴定了 mRNA水平会因酒精的作用而改变， 分子上，酒精作用的后果已经显现。结合遗传 工具和分子技术提供了一个合理的检测策略， 验证影响酒精相关性的重要遗传差异 性状 一个修改的DD分析，它有能力筛选最 表达的转录本以及鉴定插入和缺失 多态性，将在C57 Bl/6 J（B6）和DBA/2 J（D2）小鼠以及两只 作为急性酒精的QTL分析的结果创建的同类菌株 戒断首先，对B6、D2和同源菌株进行DD分析， 提供酒精响应性和菌株特异性表达差异。接下来， 微阵列将用于验证，剂量效应，时间过程， 脑区特异性将通过添加DD分离基因来定义， 用于DNA微阵列分析的额外靶点， 通过聚类分析阐明具有会聚调节的分子途径 乙醇。此外，B6、D2、B6、D3、D4、D5、D 6、D 7、D8、D9、D10、D11、D12、D13、D14、D16、D18、D19、D 这些同源菌株将对许多研究有价值， 将使用BXD策略。所有基因和分析数据将免费共享 通过存放在MGI数据库（http：/www.informaticsjax.org）和一个网络 在UT创建的详细说明方案和结果的站点;例如 见：V.Iyer，（http：//genome-www.stanford.edu/serum）。

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).