Excessive drinking and urocortin 1 neurocircuit

描述（由申请人提供）：本提案旨在成为INIA联盟“过量饮酒的神经生物学基础”的一部分，重点研究神经肽尿皮质素1 （Ucn1）。Ucn1是促肾上腺皮质激素释放因子（CRF）受体CRF1和CRF2最有效的内源性配体。大脑中Ucn1的主要来源是非神经节前Edinger-Westphal核（npEW）。npEW的主要投影区域之一是侧隔（LS）。最近的证据表明，Ucn1系统对酒精极其敏感，该系统的差异使动物对酒精摄入量的差异具有易感性，并且对该系统的操纵可以调节酒精摄入量。基于这一证据，我们假设Ucn1活性的差异是过量饮酒的重要决定因素。在这个项目中，我们建议采用合作努力来研究三个具体目标：(1)使用微阵列技术鉴定在选择性繁殖的高和低酒精消耗动物中npEWand LS中表达一致的不同基因。以下动物模型将被探索：在黑暗中选择性地过度饮酒的小鼠，在预定的液体获取过程中选择性地过度饮酒的小鼠，使用2瓶选择程序选择性地饲养的小鼠，使用2瓶选择程序选择性地饲养的大鼠，以及它们各自的控制系。基因表达的差异将通过免疫组织化学、原位杂交和定量RTPCR来证实。(2)采用三种行为模型检测Ucn1基因敲除小鼠的酒精摄入量：DID -在黑暗中过量饮酒；SHAG -由于预定的访问而过量饮酒；标准的两瓶饮用程序。我们还将使用微阵列技术来研究Ucn1敲除小鼠是否在Specific Aim 1中鉴定的基因中产生代偿。(3)检测过量饮酒动物模型中特异性Aim 1和特异性Aim 2中鉴定的基因是否在人类死后大脑npEW和LS中表达，以及它们在酒精受试者和对照组之间是否存在差异表达。鉴定基因的人类同源物将通过定量RT-PCR进行检测。综上所述，这些研究将对Ucn1神经回路及其与过度饮酒的关系提供全面透彻的分析，并可为俄勒冈州健康中心治疗酒精中毒和酒精滥用障碍的新方法的开发奠定基础。

DESCRIPTION (provided by applicant): This proposal seeks to become a part of the INIA consortium "Neurobiologial Basis of Excessive Drinking", and focuses of the neuropeptide Urocortin 1 (Ucn1). Ucn1 is the most effective endogenous ligand of both corticotropin releasing factor (CRF) receptors CRF1 and CRF2. The main source of Ucn1 in the brain is the non-preganglionic Edinger-Westphal nucleus (npEW). One of the main projection areas of npEW is the lateral septum (LS). Recent evidence indicates that the Ucn1 system is extremely sensitive to alcohol, that differences in this system predispose animals to differences in alcohol consumption, and that manipulations of this system regulate alcohol intake. Based on this evidence we hypothesize that differences in Ucn1 activity are important determinants of excessive alcohol intake. In this project we propose to apply collaborative efforts to investigate three specific aims: (1) To identify genes showing consistently different expression in npEWand LS between selectively-bred high and low alcohol consuming animals using microarray technology. Following animal models will be explored: mice selectively bred for excessive drinking in the dark, mice selectively bred for excessive drinking in the scheduled fluid access procedure, mice selectively bred using the 2-bottle choice procedure, rats selectively bred using the 2-bottle choice procedure, and their respective control lines. Differences in gene expression will be confirmed using immunohistochemistry, in situ hybridization and quantitative RTPCR. (2) To test alcohol consumption in Ucn1 knockout mice using three behavioral models: DID - excessive drinking in the dark; SHAG - excessive drinking due to scheduled access; and the standard 2-bottle drinking procedure. We will also use microarray technology to investigate whether Ucn1 knockout mice developed compensations in genes identified in Specific Aim 1. (3) To test whether genes identified in animal models of excessive alcohol consumption in Specific Aim 1 and Specific Aim 2 are expressed in npEW and LS of human post-mortem brains, and whether they are differentially expressed between alcoholic subjects and controls. Human homologues of the identified genes will be tested by quantitative RT-PCR. Taken together, these studies will provide a thorough comprehensive analysis of the Ucn1 neurocircuit and its involvement in excessive alcohol consumption, and could provide groundwork for development of new approaches for Oregon Health treatments of alcoholism and alcohol abuse disorders.