Identification of Genes for Alcohol Preference in Noninbred Rats and Mice

研究提供了相当多的证据表明，酗酒的风险受遗传因素的影响。该项目的长期目标是通过识别影响酒精偏好的基因，为酒精治疗提供新的靶点。本研究的目的是鉴定和表征选择性远交高饮酒量（HAD）和低饮酒量（LAD）大鼠以及高饮酒量偏好（HAP）和低饮酒量偏好（LAP）小鼠精细定位染色体（Chr） QTL区域的强候选基因。我们对QTL区域精细图谱的建议将通过应用一种新颖的、多点的、非常适合于分析非近交群体的血统识别方法来完成。随着QTL区间的精细定位，微阵列分析的结果和生物信息学资源的利用将使我们能够确定哪些差异表达基因在缩小的QTL区间内。这些基因的差异表达将通过实时荧光定量PCR （qRT-PCR）分析来证实。然后使用定量Western Blot分析检测强候选基因的蛋白表达差异。最后，我们将确定影响差异表达候选基因表达的编码和/或调控序列变化，并通过瞬时转染报告基因试验确定其功能意义。提出的研究将在此应用中发挥优势

Research has provided considerable evidence that the risk for alcoholism is influenced by genetic factors. The long term goal of this project is to provide novel targets for alcohol treatment by identifying genes that influence alcohol preference. The objective of this proposal is to identify and characterize the strong candidate genes in the fine mapped chromosome (Chr) QTL regions in the selectively outbred high alcohol drinking (HAD) and low alcohol drinking (LAD) rats and the high alcohol preferring (HAP) and low alcohol preference (LAP) mice. Our proposal to fine map the QTL regions will be accomplished by applying a novel, multipoint, identity by descent approach well suited to analyses of noninbred populations. Following fine mapping of the QTL intervals, the results of microarray analyses and the utilization of bioinformatic resources will allow us to determine which of the differentially expressed genes are within the narrowed QTL intervals. The differential expression of these genes will be confirmed by quantitative Real-Time PCR (qRT-PCR) analysis. Strong candidate genes will then be tested for protein expression differences by using quantitative Western Blot analyses. Finally, we will identify coding and/or regulatory sequence changes that influence the expression of the differentially expressed candidate genes and determine functional significance by transient transfection reporter assays. The proposed research in this application will take advantage of unique noninbred animal models in combination with new and innovative analyses to narrow the QTL regions and identify candidate genes that underlie alcohol preference.