Biochemical and Genetic Determinants of Differences in Alcohol Consumption

描述(由申请人提供)：INIA赠款的总体目标是定义酒精避免机制，并向其他INIA研究人员提供行为测试方面的专业知识，以评估新突变小鼠的酒精相关行为。根据我们当前INIA资助期间的数据，我们认为啮齿动物避免高浓度乙醇的原因是肝脏和大脑中促炎症细胞因子的释放和/或产生增加，这些细胞因子导致大脑中基因表达的长期变化。我们还发现，当连续饮酒并间歇每周禁欲时，最初对酒精的高度接受程度会下降，这种下降取决于遗传背景。我们开发了两种遗传小鼠模型，它们保持一致的高偏好和高消费水平(FVBxB6F1小鼠；持续酒精偏好，SAP)或形成酒精回避(NZBxB6F1小鼠；降低酒精偏好，RAP)。这些动物模型提供了比较最初饮酒、持续饮酒和戒酒发展形成的大脑基因表达谱的机会。为了验证细胞因子假说，我们将研究：1)缺乏一些促炎症细胞因子和/或其受体基因的基因敲除小鼠的酒精消耗；2)促炎症细胞因子(尤其是α-肿瘤坏死因子)和细胞因子受体拮抗剂系统处理对自愿酒精消耗的影响；3)细胞因子处理后的脑基因表达谱，以便与从酒精戒断小鼠获得的阵列数据进行比较；4)通过3个周期的戒酒和戒酒处理FVBxB6F1和NZBxB6F1杂交小鼠，并通过基因芯片分析找出关键基因；5)通过给予RNAi建立大脑区域击倒关键基因的小鼠。这项工作将使用3个INIA核心(德克萨斯阵列和信息核心，RA Harris-Pi；科罗拉多RNAi核心，W.Zawada-Pi；加利福尼亚小鼠动物模型核心，A.Roberts-Pi)以及与6个INIA调查人员的合作(A.Alcantara-奥斯汀，德克萨斯州；S.Bergeson-奥斯汀，德克萨斯州；R.D.梅菲尔德-奥斯汀；R.Davis-休斯顿；A.Ryabinin-波特兰，俄勒冈州；B.Tabakoff-奥罗拉，科罗拉多州)。

DESCRIPTION (provided by applicant): The overall objective of this INIA grant is to define mechanisms of alcohol avoidance and provide expertise in behavioral testing to other INIA investigators to evaluate ethanol-related behaviors in new mutant mice. Based on our data from the current period of INIA funding, we propose that the avoidance of high concentrations of ethanol by rodents is caused by increased release and/or production of proinflammatory cytokines in liver and brain and that these cytokines cause long-lasting changes in gene expression in the brain. We also found that initial high acceptance of alcohol can decrease when alcohol is presented continuously with intervening weekly abstinence periods, and this decrease depends upon the genetic background. We developed two genetic mouse models which maintain consistent high levels of preference and consumption (FVBxB6F1 mice; Sustained Alcohol Preference, SAP) or develop avoidance of alcohol (NZBxB6F1 mice; Reduced Alcohol Preference, RAP). These animal models provide the opportunity to compare brain gene expression profiles formed by initial alcohol consumption, sustained consumption and development of alcohol avoidance. To test the cytokine hypothesis, we will study: 1) ethanol consumption in knockout mice lacking genes for some proinflammatory cytokines and/or their receptors; 2) the effect of systemic treatment with proinflammatory cytokines (particularly a-TNF) and antagonists of cytokine receptors on voluntary ethanol consumption; 3) brain gene expression profiles after cytokine treatment to allow comparison with array data obtained from mice with ethanol avoidance; 4) treatment of FVBxB6F1 and NZBxB6F1 hybrid mice via 3 cycles of alcohol consumption and abstinence and identification of key genes by microarray analysis; 5) generation of brain regional knock-down mice of key genes by administration of RNAi. This work will use 3 INIA Cores (Texas Array and Informatics Core, RA Harris -PI; Colorado RNAi core, W. Zawada - PI; California Mouse Animal Model Core, A. Roberts - PI) and collaborations with six INIA investigators (A. Alcantara - Austin, Texas; S. Bergeson - Austin, Texas; R. D. Mayfield - Austin, Texas; R. Davis - Houston, Texas; A. Ryabinin - Portland, Oregon; B. Tabakoff - Aurora, Colorado).