m4 & m5 muscarinic acetylcholine receptor subtypes in ethanol-induced locomotion

米4

• 批准号: 7608771
• 负责人: Angela C Scibelli
• 金额: $ 4.1万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-30 至 2010-03-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7608771
• 关键词: Acute; Affect; Alcohol consumption; Alcoholism; Animal Model; Area; Behavior; Behavioral; Biological Assay; Brain region; Breeding; Chromosome Mapping; Chromosomes, Human, Pair 2; Corpus striatum structure; Data; Development; Dopamine; Dopamine D1 Receptor; Ethanol; Future; Gene Expression; Gene Silencing; Genes; Genetic; Goals; Human; Injection of therapeutic agent; Laboratory Finding; Lasers; Locomotion; Microinjections; Mission; Modeling; Motor Activity; Mus; Muscarinic Acetylcholine Receptor; Muscarinic Antagonists; National Institute on Alcohol Abuse and Alcoholism; Nucleus Accumbens; Olfactory tubercle; Pathway interactions; Pharmaceutical Preparations; Pharmacogenetics; Pharmacotherapy; Play; Predisposition; Prefrontal Cortex; Psychological reinforcement; RNA Interference; Receptor Gene; Rewards; Role; Scopolamine; Substantia nigra structure; System; Testing; Transcript; Ventral Tegmental Area; Western Blotting; alcohol abuse therapy; alcohol effect; alcohol related problem; alcohol response; alcohol sensitivity; base; brain tissue; endophenotype; human CHRM4 protein; mouse model; receptor; response; trait

DESCRIPTION (provided by applicant): Our long-term goal is to identify genes that influence sensitivity to acute locomotor stimulation to ethanol, a putative mouse model of an endophenotype (behavioral stimulation) for alcoholism. The overall goal of this proposal is to determine if the m4 and/or m5 muscarinic acetylcholine receptor (mAChR) subtype genes have a role in acute locomotor stimulation to ethanol. Genes on mouse chromosome 2 (Chr 2) influence sensitivity to ethanol-induced stimulation; within the relevant region on Chr 2 reside the m4 and m5 mAChR subtype genes. The proposed studies are intended to elucidate if differences in m4 and/or m5 gene expression are associated with predisposition to ethanol-induced stimulation, and if they are directly involved in this behavior. The FAST and SLOW lines of mice were selectively bred for high and low sensitivity to the locomotor stimulant effects of ethanol, and are therefore a sensitive genetic animal model in which to study these differences. The specific aims of this proposal are to 1.) determine whether there are specific brain regions in which the m4 and/or m5 mAChR subtype genes are differentially expressed in FAST and SLOW mice, and 2.) determine whether selective gene silencing of the m4 and/or m5 mAChR subtype genes affects acute locomotor stimulation to ethanol. Gene expression will be assessed in lasercapture-dissected brain regions using qRT-PCR and Western blot analysis. Silencing of the m4 and m5 receptor subtype genes will be accomplished by microinjecting selective RNA interference (RNAi) transcripts into the nucleus accumbens (NAc) and ventral tegmental area (VTA), respectively. It is hypothesized that FAST mice will display decreased expression of the m4 gene in the NAc and enhanced expression of the m5 gene in the VTA as compared to SLOW mice. It is also hypothesized that microinjection of m4-selective RNAi into the NAc of SLOW mice will enhance the acute locomotor stimulant response to ethanol, while microinjection of m5-selective RNAi into the VTA of FAST mice will block ethanol-induced stimulation. It is important to understand the pharmacogenetic underpinnings of acute sensitivity to ethanol because it has some value for the prediction of future alcohol use/abuse. Findings from this proposal may inform future pharmacotherapies for development in the treatment of alcoholism.

描述（由申请人提供）：我们的长期目标是鉴定影响对乙醇急性运动刺激敏感性的基因，这是一种酒精中毒内表型（行为刺激）的推定小鼠模型。本提案的总体目标是确定M4和/或M5毒蕈碱乙酰胆碱受体（mAChR）亚型基因是否在乙醇急性运动刺激中发挥作用。小鼠2号染色体（Chr 2）上的基因影响对乙醇诱导刺激的敏感性;在Chr 2上的相关区域内存在m4和m5 mAChR亚型基因。拟定的研究旨在阐明m4和/或m5基因表达的差异是否与乙醇诱导刺激的易感性相关，以及它们是否直接参与这种行为。FAST和SLOW系的小鼠被选择性地培育为对乙醇的运动刺激作用的高敏感性和低敏感性，因此是研究这些差异的敏感遗传动物模型。本提案的具体目标是：（1）确定是否存在其中m4和/或m5 mAChR亚型基因在脑中差异表达的特定脑区域， FAST和SLOW小鼠，以及2.）确定m4和/或m5 mAChR亚型基因的选择性基因沉默是否影响对乙醇的急性运动刺激。将使用qRT-PCR和Western印迹分析在激光捕获解剖的脑区域中评估基因表达。通过将选择性RNA干扰（RNAi）转录物分别显微注射到丘脑核（NAc）和腹侧被盖区（VTA）来实现m4和m5受体亚型基因的沉默。假设与SLOW小鼠相比，FAST小鼠将显示NAc中m4基因的表达降低和VTA中m5基因的表达增强。还假设将m4选择性RNAi显微注射到SLOW小鼠的NAc中将增强对乙醇的急性运动刺激反应，而将m5选择性RNAi显微注射到FAST小鼠的VTA中将阻断乙醇诱导的刺激。了解乙醇急性敏感性的药物遗传学基础是很重要的，因为它对预测未来的酒精使用/滥用有一定的价值。这项建议的发现可能会为未来酒精中毒治疗的药物治疗提供信息。