Biochemical and Genetic Determinants of Alcohol Consumption

描述（由申请人提供）：本项目基于nia - west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，删除6个INIA候选神经炎症基因中的任何一个都会减少酒精消耗，激活免疫信号会增加酒精消耗。这些数据表明，在人类酒精中毒和我们的遗传动物模型中，大脑中存在促炎信号的错误调节。我们的几个候选基因是特定toll样受体（TLR4）信号通路的一部分，我们将对其进行行为和生化研究。具体目标1：确定TLR4信号中负责促进过度饮酒的分子成分。这些研究将使用缺乏该系统关键成分的零突变小鼠。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AE1-329。具体目标2：定义小鼠过度饮酒和神经免疫激活所干扰的基因网络，并将其与人类酒精中毒的基因表达变化进行比较。这一目标还将通过测量使用消炎药减少酒精消耗的小鼠大脑中细胞因子的水平，来确定与酒精消耗调节相关的脑细胞因子的变化。Specific Aim 3是一个核心功能，将为使用RNAi、条件零突变小鼠和药理学方法的其他INIA项目提供新的INIA候选基因的行为测试。INIA相互作用：基因操作小鼠将使用来自Lasek和Homanics INIA核心的RNAi和零突变小鼠。我们将为Heberlein和Ponomarev项目提供行为测试，并给老鼠注射Ponomarev。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核心用于药物测试，Siggins和Morrisett项目用于电生理学。

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.