Genetic Studies of Alcohol Tolerance

描述（由申请人提供）：发生酒精中毒的一个重要遗传风险因素是对急性剂量酒精的不同敏感性。急性酒精反应是初始敏感性和急性功能耐受（AFT）共同作用的结果，两者都受遗传因素的影响。使用近亲繁殖的小鼠品系，我们一直在使用一种被称为快速耐受性的范式——在单次接触酒精后24小时内产生耐受性——作为研究急性酒精反应的遗传学工具。我们发现，自交系长睡眠和短睡眠小鼠品系（ILS和ISS）在发展快速耐受性的能力上存在很大差异，使用失转直反射测试（LORR）作为敏感性的衡量标准。这种菌株依赖的差异似乎至少部分是由对AFT的不同影响介导的。我们假设，快速耐受性的遗传变异是由于基线基因表达、酒精介导的基因表达影响以及基因序列和结构差异的基因型依赖差异造成的。因此，我们建议利用快速耐受性模型，利用下一代高通量深度测序技术来研究急性反应的分子和遗传基础。利用ILS和ISS衍生的LXS重组自交系（RI）小鼠品系面板，研究快速耐受性、初始敏感性和AFT的遗传学。表达谱分析将使用定量RNA测序（RNA-seq）进行，可以研究对选择性剪接以及转录物丰度的影响。提出了以下六个具体目标：1)确定LXS - RIs中对LORR反应的初始敏感性、AFT和快速耐受性之间的关系；2)为Aim 1确定的应答绘制数量性状位点（qtl）；3)对ILS和ISS的全基因组进行测序；4)在LXS - RIs的大脑中进行表达谱分析；5)绘制在Aim 4中鉴定的基因和在Aim 2中确定的行为QTL区间内发生的基因的表达QTL (eQTLs)；6)确认Aims 4和5中鉴定基因的表达结果。我们建议，这些实验的结果将提供深入了解急性酒精敏感性的遗传变异的本质。这反过来将有助于更深入地了解人类酗酒的遗传风险。

DESCRIPTION (provided by applicant): An important genetic risk factor for the development of alcoholism is differential sensitivity to an acute dose of alcohol. Acute alcohol responses are a function of the combined effects of initial sensitivity and acute functional tolerance (AFT), bot of which are influenced by genetic factors. Using inbred mouse strains, we have been using a paradigm known as rapid tolerance - tolerance that develops within 24 hrs following a single exposure to alcohol - as a tool to investigate the genetics of acute alcohol responses. We have found that the Inbred Long and Short Sleep mouse strains (ILS and ISS) differ considerably in their ability to develop rapid tolerance using the loss of righting reflex test (LORR) as the measure of sensitivity. This strain- dependent difference appears to be mediated at least partly by differential effects on AFT. We hypothesize that genetic variance in rapid tolerance occurs as a result of genotype-dependent differences in baseline gene expression, in alcohol-mediated effects on gene expression, and in differences in gene sequence and structure. Thus, we propose to exploit the rapid tolerance model to examine the molecular and genetic basis of acute responses using Next Generation high-throughput deep sequencing technologies. The genetics of rapid tolerance, initial sensitivity, and AFT will be investigated using the LXS recombinant inbred (RI) mouse strain panel which was derived from the ILS and ISS. Expression profiling will be conducted using quantitative RNA sequencing (RNA-seq) with which it is possible to investigate effects on alternative splicing as well as on transcript abundance. The following six Specific Aims are being proposed: 1) determine relationships between initial sensitivity, AFT, and rapid tolerance for the LORR response in the LXS RIs; 2) map quantitative trait loci (QTLs) for the responses determined in Aim 1; 3) sequence the full genomes of the ILS and ISS; 4) conduct expression profiling in the brains of the LXS RIs; 5) map expression QTLs (eQTLs) for genes identified in Aim 4 and for genes that occur within the behavioral QTL intervals determined in Aim 2; and 6) confirm expression results for genes identified in Aims 4 and 5. We propose that the results of these experiments will offer insight into the nature of genetic variance for acute alcohol sensitivity. This in turn will contribute to a deeper understanding of genetic risk for human alcoholism. PUBLIC HEALTH RELEVANCE: The initiation and maintenance of alcoholism is influenced by both environmental and genetic factors. This project aims to identify genes that influence variation in acute alcohol sensitivity, a trait that is thought to contribute to genetic risk for alcoholism. Such knowledge is essential for a complete understanding of the molecular basis of alcoholism and for the development of new or improved strategies for its treatment.