Excessive drinking and urocortin 1 neurocircuit

过量饮酒与尿皮质素1神经回路

• 批准号: 7214426
• 负责人: Andrey E Ryabinin
• 金额: $ 24.6万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-30 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7214426
• 关键词: alcoholic beverage consumption; alcoholism /alcohol abuse; behavior test; brain septal area; cooperative study; corticotropin releasing factor; ethanol; genetic models; genetically modified animals; hormone regulation /control mechanism; human tissue; immunocytochemistry; in situ hybridization; laboratory mouse; laboratory rat; microarray technology; neuroendocrine system; neurogenetics; neuroregulation; polymerase chain reaction; postmortem; psychological models

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.

描述（由申请人提供）：该提案旨在成为INIA联盟“过度饮酒的神经生物学基础”的一部分，并将神经肽尿素素1（UCN1）的重点（UCN1）组成。 UCN1是皮质激素释放因子（CRF）受体CRF1和CRF2的最有效的内源配体。大脑中UCN1的主要来源是非限制的Edinger-westphal核（NPEW）。 NPEW的主要投影区域之一是侧隔隔隔（LS）。最近的证据表明，UCN1系统对酒精非常敏感，该系统的差异使动物易于饮酒的差异，并且该系统的操纵调节酒精摄入量。基于这一证据，我们假设UCN1活性的差异是饮酒过量的重要决定因素。在这个项目中，我们建议采用协作努力来研究三个特定目标：（1）使用微阵列技术鉴定在有选择性的高和低饮酒动物之间显示出Npewand ls持续不同表达的基因。将探讨以下动物模型：在黑暗中有选择性地育出了用于过量饮用的小鼠，在预定的液体进入程序中有选择地育出了过量饮用的小鼠，使用2瓶选择程序选择性地育出了小鼠，使用2瓶选择程序选择性地育出了大鼠，并使用2瓶选择过程饲养。基因表达的差异将使用免疫组织化学，原位杂交和定量RTPCR确认。 （2）使用三种行为模型在UCN1敲除小鼠中测试饮酒：DID - 在黑暗中饮酒过多；粗毛 - 由于预定的通道而导致过多的饮酒；以及标准的2瓶饮酒程序。我们还将使用微阵列技术来研究在特定目的1中鉴定出的基因中的UCN1敲除小鼠。（3）测试在特定目标1中过度饮酒的动物模型中是否鉴定出的基因和特定目标2在人类邮政brains的NPEW和LS中表达了人类邮政的NPEW和LS LS，以及它们是否在酗酒者和对照组中差异表达。鉴定基因的人类同源物将通过定量RT-PCR进行测试。综上所述，这些研究将提供对UCN1神经电路及其参与过度饮酒的全面分析，并可以为开发俄勒冈州健康治疗方法的新方法提供基础。