BACKCROSS STRATEGY TO IDENTIFY ALCOHOL RELATED B6 QTLS

识别酒精相关 B6 QTLS 的回交策略

• 批准号: 6163744
• 负责人: LEE M SILVER
• 金额: $ 21.17万
• 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 2002-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6163744
• 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.

我们建议使用一种互交-回交育种方案， 选择性基因分型，以识别、定位和表征基因 C57BL/6表达的高度酒精偏好特征 小鼠在我们迄今为止的研究中，我们已经确定并定位了一种 先前未预测的酒精偏好位点-Alcp1-处于高水平 显著性（P <0.00018）为一条小鼠染色体上的8cM间隔。 第二个酒精偏好基因座-Alcp2-已被初步定位 在第二条染色体上（P <0.0013）。这两个位点都与基因紧密相连 参与血清素活动。本研究的具体目的 建议如下：1.酒精偏好相关基因座的鉴定 B6小鼠表达的性状。来自（B6 X DBA）X B6的额外后代 将筛选回交以获得总共300只动物， 表现出高度酒精偏好的特征。对这些进行分层分析 带有微卫星标记的动物将使我们能够识别和 绘制显示与酒精至少60%关联的QTL图谱 偏爱类将定义多个QTL之间的相关性， 用于评估遗传相互作用的各种模型。第二 将分析回交以评估结果的一般性 用B6和DBA获得。2.候选人的遗传和分子分析 酒精偏好基因候选人的降钙素相关基因已经 确定的，以及由特定的绘图研究提出的任何其他建议 目标1，将检测与Alcp1、Alcp2或其他 未被覆盖的Alcp基因座。在这种遗传中存活下来的候选基因 将检测编码序列和RNA转录 亲本B6和DBA等位基因之间的差异。3.进一步神经化学 以及对嗜酒小鼠的行为研究。协同 巴特·霍贝尔教授将对喜欢喝酒的老鼠进行分析， 额外的成瘾行为特征的表达和特定的 神经化学物质预测发挥作用，在这些特征的基础上， 目标1和2的遗传结果。 如果任何形式的其他关联 观察到，累积的数据将用于确定 基因型-表型关系的特异性。