Indiana Gene Array Core

描述（由申请人提供）：印第安纳基因阵列核心的总体目标是与INIA研究人员合作，了解涉及过度饮酒发展的扩展杏仁核中基因表达的差异。我们的假设是，基因表达的先天差异和对酒精的反应以及涉及酒精暴露或戒酒的范式（基因-环境相互作用）的差异导致了过度饮酒的发展。印第安纳基因阵列核心将专注于使用Affymetrix微阵列来测量各种治疗后大鼠大脑选定区域和果蝇突变体的基因表达；主要合作者将是McBride (U01), Ryabinin （U01）和Davis （U01）。这些研究建立在我们最初资助期间的工作基础上；在同一实验室（由许多相同的技术人员）进行后续研究将减少技术差异，并允许更有力的元分析。我们将与研究人员一起进行实验设计，从合作研究人员提供的大鼠脑组织中提取和纯化RNA，执行所有涉及GeneChip实验的质量控制和生物化学，提取数据并协助分析和生物信息学。这个核心将被安置在由PI指导的医学基因组学中心，该中心拥有所有必要的设备，并进行大量Affymetrix基因芯片实验，重点是质量控制。我们将扩展我们的服务，包括对我们处理的微阵列的关键发现进行定量RT-PCR确认，并将向INIA的其他成员提供此类服务。我们还将为INIA成员提供在微阵列实验中检测到的导致基因表达差异的顺式作用元件的分析；我们已经开发了基于模型的技术，不需要事先了解结合位点。该核心将以高效和经济的方式提供高质量的数据。我们已经与用户建立了牢固的关系，这将有利于整个项目。

DESCRIPTION (provided by applicant): The overall aim of the Indiana Gene Array Core is to collaborate with INIA investigators to understand the differences in gene expression in the extended amygdala that are involved in the development of excessive drinking. Our hypothesis is that both innate differences in gene expression and differences in response to alcohol and to paradigms involving exposure to alcohol or its withdrawal (gene-environment interactions) contribute to the development of excessive alcohol drinking. The Indiana Gene Array Core will focus on the use of Affymetrix microarrays to measure gene expression in selected regions of rat brains and in Drosophila mutants after various treatments; primary collaborators will be McBride (U01), Ryabinin (U01) and Davis (U01). These studies build upon our work during the initial grant period; carrying out the follow-up studies in the same laboratory (with many of the same technical personnel) will reduce technical variations and allow more powerful metaanalyses. We will work with investigators on experimental design, extract and purify RNA from rat brain tissues provided by the collaborating investigators, carry out all of the quality control and biochemistry involved in GeneChip experiments, extract the data and assist with analyses and bioinformatics. This core will be housed in the Center for Medical Genomics, directed by the PI, which has all of the necessary equipment and carries out a high volume of Affymetrix GeneChip experiments with a focus on quality control. We will extend our services to include quantitative RT-PCR confirmation of key findings from the microarrays that we process, and will offer such services to other members of INIA. We will also provide members of INIA analyses of cis-acting elements that contribute to the differences in gene expression detected in the microarray experiments; we have developed model-based techniques that do not require prior knowledge of binding sites. This Core will provide high quality data in an efficient and economical way. We already have strong relatioinships with the users, which will benefit the overall project.