Mapping and Microarray Gene Expression Analysis in a Model of Excessive Drinking

描述(申请人提供)：饮酒是一个复杂的特征，因此受到多种遗传和环境因素的影响。一次检查一个基因的影响忽略了可能的相互作用的重要性(例如，上位性、等位基因内的超显性；见Phillips&Belnuap，2002)。我们建议利用子代杂交的方法来定位导致过量饮酒的基因组。已发现C57BL/6J(B6)x FVB/NJ(FVB)F1小鼠甚至比高酒精偏好的B6品系消耗更多的酒精(Blednov等人，2005年)。我们将利用这一发现，通过结合数量性状基因座(QTL)定位和微阵列基因表达分析来确定行为(BQTL)和表达QTL(EQTL)的共定位，从而解决过度饮酒的遗传模式。在具体目标1中，将使用B6FVBF2定位过度自愿乙醇消费的bQTL。F2将被用来确定每个QTL的遗传模式(加性、完全显性、超显性、上位性)，也可以用成对组合。在特定目标2中，将使用F3-F4个体进行脑区特定基因表达分析。根据他们的基因预测的高饮酒率或低饮酒率的个体将被选择进行微阵列分析。这些数据将接受eQTL分析。这将使我们能够识别定位到共同染色体区域的bQTL和eQTL，为影响饮酒特性的特定基因(S)提供证据。将从INIA神经回路小组确定的目标组织中选择要研究的大脑区域。在具体目标3中，将测量可能的遗传相关反应，以探索极端饮酒特征与其他可能的遗传关系，如酒精戒断、条件性味觉厌恶、戒断诱导饮酒、在黑暗中饮酒、酒精接受和酒精条件位置偏好。随着我们获得影响高饮酒性状的基因特定位置的证据，我们将根据先前的QTL定位数据选择其他性状进行检查，这些数据已经确定了这些性状与我们确定的位置的关联。未来的工作还将集中在我们的联合bQTL：eQTL分析所涉及的最重要的候选基因上。

DESCRIPTION (provided by applicant): Ethanol drinking is a complex trait, and thus influenced by multiple genetic and environmental factors. Examining the influence of one gene at a time ignores the importance of possible interactions (e.g., epistasis, intra-allelic overdominance; see Phillips & Belknap, 2002). We propose to utilize a filial cross approach for the mapping of gene sets that result in excessive alcohol intake. C57BL/6J (B6) x FVB/NJ (FVB) F1 mice have been found to consume more ethanol than even the high ethanol preference B6 strain (Blednov et al., 2005). We will take advantage of this finding to resolve mode of inheritance for excessive drinking by combining quantitative trait locus (QTL) mapping with microarray gene expression analysis to identify colocalization of behavioral (bQTL) and expression QTL (eQTL). In Specific Aim 1, bQTL for excessive voluntary ethanol consumption will be mapped using the B6FVBF2. The F2 will be used to determine the mode of inheritance (additive, fully dominant, overdominant, epistatic) for each QTL taken singly and also in pairwise combinations. In Specific Aim 2, F3-F4 individuals will be used for brain region specific gene expression analyses. Individuals predicted by their genotype to be high or low drinking individuals will be selected for microarray profiling. These data will be subjected to eQTL analyses. This will allow us to identify bQTL and eQTL that are mapped to common chromosomal regions providing evidence of the specific gene(s) influencing the drinking trait. Brain regions to be studied will be selected from target tissues identified by the INIA Neurocircuitry group. In Specific Aim 3, possible genetically correlated responses will be measured to explore putative genetic relationships between the extreme drinking trait and others such as ethanol withdrawal, conditioned taste aversion, withdrawal induced drinking, drinking in the dark, ethanol acceptance, and ethanol conditioned place preference. As we obtain evidence for the specific locations of genes that influence the high drinking trait, other traits will be chosen for examination based on previous QTL mapping data that have identified associations of those traits with the locations we identify. Future work will also be focused on the most significant candidate genes implicated by our combined bQTL:eQTL analyses.