Biochemical and Genetic Determinants of Alcohol Consumption

描述（由申请人提供）：该项目基于INIA-West研究，该研究显示了酒精摄入动物模型和人类酗酒者大脑中神经免疫基因表达的变化。我们发现，六个INIA候选神经炎症基因中的任何一个的缺失都会减少酒精消耗，而免疫信号的激活会增加酒精消耗。这些数据表明，在人类酒精中毒和我们的遗传动物模型中，大脑中的促炎信号存在失调。我们的几个候选基因是一个特定的Toll样受体（TLR 4）信号通路的一部分，我们将研究行为和生化。具体目标1：定义负责促进过量饮酒的TLR 4信号传导的分子组分。这些研究将使用缺乏该系统关键组分的无效突变小鼠。神经炎症信号也是酒精中毒药物开发的潜在目标，我们将测试三种抗炎药物：米诺环素，吡格列酮和AE 1 -329。具体目标2将：定义小鼠中被过量饮酒和神经免疫激活干扰的基因网络，并将其与人类酒精中毒中的基因表达变化进行比较。该目标还将通过测量用减少酒精消耗的抗炎药物治疗的小鼠脑中的细胞因子水平来定义与酒精消耗调节相关的脑细胞因子的变化。具体目标3是一个核心功能，将使用RNAi、条件无效突变小鼠和药理学方法为其他INIA项目提供新INIA候选基因的行为测试。INIA相互作用：小鼠中的遗传操作将使用来自Lasek和Homanics INIA核心的RNAi和无效突变小鼠。我们将为Heberlein和Ponomarev项目提供行为测试，并为Ponomarev治疗小鼠。我们将与Mayfield和Ponomarev项目合作，比较我们的基因表达谱数据（人类和小鼠），Roberts/Kosten核心药物测试以及Siggins和Morrisett项目的电生理学。

DESCRIPTION (provided by applicant): This project is based on INIA-West studies showing changes in neuroimmune gene expression in animal models of alcohol intake and in brain of human alcoholics. We found that deletion of any of six INIA candidate neuroinflammatory genes decreased alcohol consumption and activation of immune signaling increased alcohol consumption. These data suggest that in human alcoholism and in our genetic animal models there is a misregulation of pro-inflammatory signaling in brain. Several of our candidate genes are part of a specific toll-like receptor (TLR4) signaling pathway that we will study behaviorally and biochemically. Specific Aim 1 will: Define the molecular components of TLR4 signaling that are responsible for promotion of excessive alcohol consumption. These studies will use null mutant mice lacking key components of this system. Neuroinflammatory signaling is also a potential target for medication development for alcoholism and we will test three anti-inflammatory drugs: Minocycline, Pioglitazone and AE1-329. Specific Aim 2 will: Define the gene networks that are perturbed by excessive alcohol consumption and neuroimmune activation in mouse and compare these to gene expression changes in human alcoholism. This aim will also define changes in brain cytokines related to regulation of alcohol consumption by measuring cytokine levels in brain of mice treated with anti-inflammatory drugs which reduce alcohol consumption. Specific Aim 3 is a Core function that will provide behavioral testing of new INIA candidate genes for other INIA projects using RNAi, conditional null mutant mice and pharmacological approaches. INIA Interactions: Genetic manipulation In mice will use RNAi and null mutant mice from the Lasek and Homanics INIA cores. We will provide behavioral testing for the Heberlein and Ponomarev projects and treated mice to Ponomarev. We will collaborate with the Mayfield and Ponomarev projects to compare our data for gene expression profiling (human and mouse), the Roberts/Kosten cores for medication testing and the Siggins and Morrisett projects for electrophysiology.