INIA: Chimeric Analysis of Alcohol & Stress Interactions

INIA：酒精的嵌合分析

• 批准号: 6694113
• 负责人: KRISTIN M HAMRE
• 金额: $ 14.05万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-02-01 至 2005-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6694113
• 关键词: NMDA receptors; alcoholic beverage consumption; alcoholism /alcohol abuse; animal breeding; behavior test; behavioral /social science research tag; behavioral genetics; biotechnology; drug withdrawal; electrostimulus; embryo /fetus; fusion gene; gene dosage; gene environment interaction; genetic models; genetically modified animals; genotype; histology; laboratory mouse; model design /development; phenotype; protein structure; stress; substance abuse related behavior; tissue mosaicism

DESCRIPTION (provided by applicant): Alcoholism and alcohol-related phenotypes have a strong genetic component. Numerous candidate genes have been hypothesized to be critical in determining these phenotypes including NMDA, serotonin and GABA systems. Typically, the way to demonstrate which of these candidate molecules are in fact responsible for the phenotype is through the use of knock-out mice. In some instances, the ability to address this issue is complicated by the fact that the mutation is embryonic or neonatal lethal making it impossible to ascertain the role of the gene in most behavioral phenotypes, particularly ethanol-related behaviors. One method of eliminating problem is through the use of chimeric mice. Chimeric mice are made by mixing mutant and wild-tax e many instances, the presence of wild-type cells is able to eliminate the lethality allowing for the analysis of animals. NMDA receptors have been hypothesized to mediate multiple alcohol-related effects including increased alcohol consumption and severity of withdrawal effects. Additionally, NMDA receptors have been shown to be important in responses to stress. The NMDA receptor complex is made up of multiple subunits and it is unclear which of the subunits are critical for these effects. Two subunits that have been hypothesized to be important are the NRI and NR2B subunits. Knock-outs of both of these subunits are embryonic lethal and thus, the definitive resolution of the role of each receptor is unknown. To determine the role of these receptors, two different chimeric combinations will be make: NRI-/-<-> wild-type and NR2B-/-<-> wild-type chimeric mice. Adult chimeric mice will be tested on both a two-bottle choice test of ethanol consumption and in a response to stress test using foot shock evaluated in an elevated plus maze. Subsequently, the brains of each chimeric mouse will be analyzed histologically to determine 1) if there is a gene dosage effect i.e. if the behavior is more abnormal in chimeric mice with a higher percentage of mutant cells, and 2) if there is a greater importance of knock-out cells in some brain regions versus others. This data will provide insights into the role of the NR1 and NR2B genes in these phenotypes.

描述（由申请人提供）： 酒精中毒和与酒精有关的表型具有很强的遗传成分。 许多候选基因已被认为对确定至关重要 这些表型在内，包括NMDA，5-羟色胺和GABA系统。通常， 证明这些候选分子中的哪个实际上是负责任的方法 因为表型是通过使用敲除小鼠。在某些情况下， 解决这个问题的能力使突变是 胚胎或新生儿致死，使得无法确定 在大多数行为表型中，尤其是与乙醇相关的行为中的基因。 消除问题的一种方法是使用嵌合小鼠。 嵌合小鼠是通过混合突变体和野生税的许多实例制成的 野生型细胞的存在能够消除允许的致死性 动物的分析。 NMDA受体已被假设介导 多种与酒精有关的影响，包括增加酒精消耗和 戒断效果的严重程度。此外，已经显示了NMDA受体 对于压力的反应很重要。 NMDA受体复合物被组成 多个亚基，尚不清楚哪个亚基对于 这些影响。假设重要的两个亚基是 NRI和NR2B亚基。这两个亚基的敲除都是胚胎 致命，因此，每个受体的作用的明确分辨率是 未知。为了确定这些受体的作用，两个不同的嵌合 组合将是：NRI - / - <->野生型和NR2B - / - <->野生型 嵌合小鼠。成年嵌合小鼠将在两个瓶中进行测试 乙醇消耗的测试以及使用脚冲击的应力测试的响应 在高架的迷宫中评估。随后，每个嵌合体的大脑 将小鼠在组织学上分析以确定1）如果存在基因 剂量效应，即，如果嵌合小鼠的行为更为异常 突变细胞的百分比较高，2）如果有更大的重要性 某些大脑区域与其他大脑区域的敲除细胞。这些数据将提供 洞悉NR1和NR2B基因在这些表型中的作用。