Identification of Genes for Alcohol Preference:Rats/Mice

研究提供了相当多的证据表明，酗酒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繁殖的高酒和低酒(HAD)大鼠和高酒偏好(HAP)和低酒偏好(LAP)小鼠的精细定位染色体(Chr)QTL区域寻找和鉴定强候选基因。我们提出的精细定位QTL区域的建议将通过应用一种新的、多点、按血统识别的方法来实现，该方法非常适合于非自交系群体的分析。在QTL区间的精细定位之后，微阵列分析的结果和生物信息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定量聚合酶链式反应(qRT-PCR)分析来证实。然后，将使用定量Western Blot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，并通过瞬时转染报告分析确定功能意义。本应用程序中提议的研究将利用 独特的非近亲繁殖动物模型与新的和创新的分析相结合，以缩小QTL区域并识别导致饮酒偏好的候选基因。

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.