Identification of natural variants that influence responses to ethanol in C. elegans

项目总结 人类的基因变异导致个人患上酒精使用障碍的可能性。这个 因此，影响酗酒倾向的基因变异是一个重要的研究目标，但它一直是 很难确定具体的责任基因。在动物模型中的实验室研究在以下方面非常有用 阐明酒精(乙醇)的分子药理学，但实验室衍生的遗传操作 很少对在野生种群中观察到的自然发生的遗传变异进行建模。因此，预测 相关的人类等位基因变异一直很难。在这里，我们研究在野外发现的自然等位基因变异。 线虫线虫的菌株，以确定在野外和 可以调节影响对乙醇的生理反应的通路的功能。线虫是一种 与人类相关的重要模式物种；在 蠕虫和人类神经系统功能的机制，以及影响酒精反应的基因 蠕虫的行为也会影响人类患上酒精使用障碍的可能性。 这项合作建议将两个实验室的不同专业知识结合在一起。我们利用了一个 独一无二的资源，来自四个遗传差异的野生菌株的重组自交系(RIL)用于鉴定 可以调节乙醇效应的自然等位基因变异。我们已经证明了亲本野生型 菌株和衍生的rIL在不同的行为反应乙醇中表现出一系列的表型。我们会 利用处理线虫模型的效率和简便性进行高通量分析，以 将确定改变对乙醇的行为和/或转录反应的遗传变异。我们会直接 检验由数量性状基因座鉴定的候选酒精反应等位基因变异的因果性质 通过使用基因编辑技术(CRISPR-Cas9)将等位基因引入 携带不同的等位基因。比较和对比遗传变异对不同基因的影响的能力 对乙醇的反应为我们的分析带来了进一步的力量。我们将确定哪些基因会影响 在每种行为中的生理反应是唯一的，第二，我们将识别影响 不同酒精反应表型的行为反应。我们还将评估等位基因对 在行为反应中调节对酒精反应的转录。这些不同的分析可以告诉我们 对乙醇的这些不同反应背后的分子机制。 总而言之，这些研究将为人类神经遗传学提供更具体和更一般的新见解。 乙醇。我们将确定已知或新的生物途径中的遗传变异程度 调节或调节乙醇的作用可以改变对乙醇的反应以及变化的程度 这些基因在野外是可以容忍的。这些数据将有助于我们理解人类易受虐待的责任 酒精。

PROJECT SUMMARY Genetic variation in humans contributes to an individual's likelihood to develop an alcohol use disorder. The genetic variation that influences alcohol abuse liability is therefore an important target for study, but it has been difficult to identify specific liability genes. Laboratory studies in animal models have been extremely useful in elucidating the molecular pharmacology of alcohol (ethanol), but laboratory derived genetic manipulations rarely model the naturally occurring genetic variation that is observed in wild populations. As such, predicting relevant human allelic variation has been difficult. Here, we study the natural allelic variation found in wild strains of the the nematode worm Caenorhabditis elegans to identify alleles that are tolerated in the wild and can modulate the function of pathways that impact physiological responses to ethanol. C. elegans is an important model species with demonstrated relevance to humans; there is striking conservation between the machinery of nervous system function in worms and humans, and genes that influence ethanol response behaviors in worms also influence the likelihood to develop alcohol use disorders in humans. This collaborative proposal brings together the diverse expertise of two laboratories. We take advantage of a unique resource, recombinant inbred lines (RILs) derived from four genetically diverse wild strains to identify natural allelic variation that can modulate the effects of ethanol. We have shown that the parent wild-type strains and the derived RILs display a range of phenotypes in different behavioral responses ethanol. We will exploit the efficiency and ease of manipulating the C. elegans model to carry out high throughput analyses that will identify genetic variation that alters behavioral and/or transcriptional responses to ethanol. We will directly test the causal nature of candidate ethanol response allelic variants, identified by quantitative trait locus mapping, through the use of gene editing techniques (CRISPR-Cas9) to introduce the allele into strains that carry different alleles. The ability to compare and contrast the influence of genetic variants on different responses to ethanol brings further power to our analyses. We will identify the genes that impact the physiological responses in each behavior uniquely, and second, we will identify genes that affect the behavioral responses across ethanol response phenotypes. We will also assess the impact of alleles that modulate transcription in response to ethanol on behavioral responses. These different analyses can inform us of the molecular mechanisms underlying these different responses to ethanol. Together, these studies will provide both specific and more general novel insights into the neurogenetics of ethanol. We will establish the degree to which genetic variation in known or novel biological pathways that mediate or modulate the effect of ethanol can change responses to ethanol and the degree to which variation in those genes is tolerated in the wild. These data will inform our understanding of human liability to abuse alcohol.