Identification of Genes for Alcohol Preference in Noninbred Rats and Mice

研究提供了大量证据表明，酒精中毒的风险受到遗传因素的影响。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。本研究的目的是在选择性远交的高酒精饮用（HAD）和低酒精饮用（LAD）大鼠以及高酒精偏好（HAP）和低酒精偏好（HAP）小鼠的染色体（Chr）QTL精细定位区域中鉴定和表征强候选基因。我们的建议，精细定位QTL区域将通过应用一种新的，多点，身份的后裔的方法非常适合于非近交群体的分析。在精细定位QTL间隔后，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的基因在缩小的QTL间隔内。将通过定量实时PCR（qRT-PCR）分析确认这些基因的差异表达。然后通过使用定量Western印迹分析来测试强候选基因的蛋白质表达差异。最后，我们将确定编码和/或调节序列的变化，影响差异表达的候选基因的表达，并确定功能的意义，通过瞬时转染报告分析。本申请中的拟议研究将利用 独特的非近交动物模型，结合新的和创新的分析，以缩小QTL区域，并确定候选基因的基础酒精偏好。

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.