BACKCROSS STRATEGY TO IDENTIFY ALCOHOL RELATED B6 QTLS

识别酒精相关 B6 QTLS 的回交策略

• 批准号: 2882038
• 负责人: LEE M SILVER
• 金额: $ 20.35万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-03-01 至 2001-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2882038
• 关键词: Mus musculus; alcoholism /alcohol abuse; alleles; animal breeding; animal genetic material tag; behavior test; behavioral /social science research tag; behavioral genetics; genetic mapping; genetic markers; genetic transcription; genome; genotype; linkage mapping; neurochemistry; nucleic acid sequence; phenotype; polymerase chain reaction; preference; single strand conformation polymorphism

We propose the use of an intercross-backcross breeding protocol with selective genotyping to identify, map, and characterize the genes responsible for the high alcohol preference trait expressed by C57BL/6 mice. In our studies to date, we have identified and localized a previously-unpredicted alcohol preference locus - Alcp1 - at a high level of significance (P <0.00018) to an 8 cM interval on one mouse chromosome. A second alcohol preference locus - Alcp2 - has been tentatively localized on a second chromosome (P <0.0013). Both of these loci map close to genes involved in serotonin activity. The specific aims of this research proposal are: 1. Identification of loci involved in the alcohol preference trait expressed by B6 mice. Additional offspring from the (B6 X DBA) X B6 backcross will be screened to obtain a total set of 300 animals that express the high alcohol preference trait. A stratified analysis of these animals with sets of microsatellite markers will allow us to identify and map QTLs showing an association of at least 6O% with the alcohol preferring class. Correlations among multiple QTLs will be defined and used to evaluate various models of genetic interactions. A second backcross will be analyzed to assess the generality of the results obtained with B6 and DBA. 2. Genetic and molecular analysis of candidate alcohol preference genes. Candidate serotonin-related genes already identified, and any others suggested by the mapping studies of specific aim 1, will be tested for co-segregation with Alcp1, Alcp2 , or further Alcp loci that are uncovered. Candidate genes that survive this genetic test will be assayed for coding sequence and RNA transcriptional differences between parental B6 and DBA alleles. 3. Further neurochemical and behavioral studies of alcohol-preferring mice. In collaboration with Professor Bart Hoebel, alcohol-preferring mice will be analyzed for the expression of additional addictive behavior traits and for specific neurochemicals predicted to play a role in these traits based on the genetic results of aims 1 and 2. If additional associations of any kind are observed, the accumulated data will be used to determine the specificity of genotype-phenotype relationships.

我们提出使用与 选择性基因分型以识别，映射和表征基因 负责C57BL/6表达的高酒精偏好特征 老鼠。在迄今为止的研究中，我们已经确定并局部 以前未经预测的酒精偏好基因座-Alcp1-高水平 一只小鼠染色体的显着性（p <0.00018）至8 cm的间隔。 第二个酒精偏好基因座-Alcp2-已暂时定位 在第二个染色体上（p <0.0013）。这两个基因座图都接近基因 参与5-羟色胺活性。这项研究的具体目的 建议是：1。鉴定酒精偏爱的基因座 由B6小鼠表达的特征。 （B6 x DBA）X B6的其他后代 将筛选反向交叉以获取总计300只动物 表达高酒精偏好特征。对这些的分层分析 具有一组微卫星标记的动物将使我们能够识别和 地图QTL显示至少6o％与酒精的关联 喜欢课。将定义多个QTL之间的相关性，并 用于评估各种遗传相互作用模型。第二 将分析反向交叉以评估结果的普遍性 用B6和DBA获得。 2。候选者的遗传和分子分析 酒精偏好基因。候选5-羟色胺相关基因已经 确定的，以及针对特定的映射研究所建议的 AIM 1将测试与Alcp1，Alcp2或进一步的共隔离 发现的ALCP基因座。生存这种遗传的候选基因 测试将用于编码序列和RNA转录的测试 父母B6和DBA等位基因之间的差异。 3。进一步的神经化学 和饮酒小鼠的行为研究。与 Bart Hoebel教授，将分析饮酒小鼠的小鼠 表达其他上瘾的行为特征和特定的 神经化学物质预测在这些特征中起作用 目标1和2的遗传结果。如果任何类型的其他关联 观察到，累积数据将用于确定 基因型 - 表型关系的特异性。