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.

我们建议使用异交-回交育种方案 对基因进行选择性基因分型以识别、定位和表征基因 对C57BL/6表现出的高酒精偏好性状负责 老鼠。在我们到目前为止的研究中，我们已经确定并定位了一个 此前未预测的酒精偏好基因Alcp1处于高水平 对一条小鼠染色体上8厘米的间隔有显著意义(P&lt；0.00018)。 第二个酒精偏好基因Alcp2已初步定位 在第二条染色体上(P&lt；0.0013)。这两个基因座都位于基因附近。 参与5-羟色胺的活动。这项研究的具体目的 建议如下：1.确定与酒精偏好有关的基因座 B6小鼠所表达的特征。(B6 X DBA)X B6的额外后代 回交将被筛选，以获得总共300只动物，这些动物 表现出较高的酒精偏好特征。对这些问题的分层分析 拥有一组微卫星标记的动物将使我们能够识别和 MAP QTL显示至少60%与酒精的关联 更喜欢班级。将定义多个QTL之间的相关性并 用于评估各种遗传交互作用模型。一秒钟 将分析回交以评估结果的一般性 使用B6和DBA获得。2.候选人的遗传和分子分析 酒精偏好基因。候选5-羟色胺相关基因已经存在 以及由特定的地图研究提出的任何其他建议 目标1将测试与Alcp1、Alcp2或更高级别的共隔离 已发现的ALCP基因座。在这种基因中存活下来的候选基因 测试将检测编码序列和RNA转录 亲本B6和DBA等位基因的差异。3.进一步的神经化学 以及对偏爱酒精的小鼠的行为研究。与 Bart Hoebel教授，偏爱酒精的小鼠将被分析 其他成瘾行为特征的表现以及对特定行为的影响 神经化学物质被预测在这些特征中发挥作用，基于 目标1和目标2的遗传结果。如果任何类型的额外关联 ，则将使用累积的数据来确定 基因型-表型关系的特异性。