BACKCROSS STRATEGY TO IDENTIFY ALCOHOL RELATED B6 QTLS

识别酒精相关 B6 QTLS 的回交策略

• 批准号: 2376088
• 负责人: LEE M SILVER
• 金额: $ 18.82万
• 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/2376088
• 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.

我们建议使用间回交育种方案 选择性基因分型以识别、绘制和表征基因 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，我们已经确定并定位了一个 先前未预测的酒精偏好基因座 - Alcp1 - 处于高水平 对一条小鼠染色体上的 8 cM 间隔具有显着性（P <0.00018）。 第二个酒精偏好基因座 - Alcp2 - 已初步定位 在第二条染色体上（P <0.0013）。这两个位点都靠近基因 参与血清素活性。本研究的具体目的 建议是： 1. 鉴定与酒精偏好有关的位点 B6小鼠表达的性状。 (B6 X DBA) X B6 的其他后代 将筛选回交以获得总共 300 只动物 表达高度酒精偏好特征。对这些内容进行分层分析 具有微卫星标记组的动物将使我们能够识别和 地图 QTL 显示至少 6O% 与酒精相关 更喜欢班级。将定义多个 QTL 之间的相关性并 用于评估遗传相互作用的各种模型。一秒钟 将分析回交以评估结果的普遍性 通过 B6 和 DBA 获得。 2. 候选者的遗传和分子分析 酒精偏好基因。候选血清素相关基因已经存在 确定的，以及特定映射研究建议的任何其他内容 目标 1，将测试与 Alcp1、Alcp2 或进一步的共分离 未发现的 Alcp 位点。在此遗传中存活下来的候选基因 测试将分析编码序列和RNA转录 亲本 B6 和 DBA 等位基因之间的差异。 3. 进一步的神经化学 以及嗜酒小鼠的行为研究。与合作 Bart Hoebel 教授，将对嗜酒小鼠进行分析 表达额外的成瘾行为特征和特定的 根据预测，神经化学物质在这些特征中发挥作用 目标 1 和 2 的遗传结果。如果有任何类型的其他关联 观察到，累积的数据将用于确定 基因型-表型关系的特异性。