A screen to identify mutations that disrupt alcohol tolerance in Drosophila

筛选破坏果蝇酒精耐受性的突变

• 批准号: 7701455
• 负责人: Roseanna Ramazani
• 金额: $ 3.43万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-09 至 2010-09-08
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7701455
• 关键词: Affect; Affinity; Alcohol consumption; Alcoholic Intoxication; Alcoholism; Alcohols; Behavior; Behavioral; Biological Assay; Biological Models; Candidate Disease Gene; Chromosome Deletion; Chromosomes; Collection; Complex; Development; Diagnostic and Statistical Manual; Disease; Drosophila genus; Drosophila melanogaster; Environment; Ethanol; Exhibits; Exposure to; Fruit; Future; Gene Expression; Genes; Genetic; Genetic Models; Genetic Screening; Genetic Transcription; Genotype; Goals; Head; Health; Human; Lead; Learning; Link; Literature; Mammals; Measures; Messenger RNA; Methods; Modification; Molecular; Mutation; Organism; Pattern; Phase; Phenotype; Population; Qualifying; Research; Reverse Transcriptase Polymerase Chain Reaction; Scientist; Screening procedure; Source; Speed; Substance abuse problem; System; Testing; Time; Training; Work; addiction; alcohol behavior; alcohol content; alcohol effect; alcohol exposure; alcohol research; alcohol response; alcoholism prevention; base; design; experience; fly; gene induction; genetic manipulation; insight; mutant; preference; social; tool

DESCRIPTION (provided by applicant): Despite a wide and varied body of literature, many unanswered questions remain in the field of ethanol research. Research into ethanol's effects with Drosophila melanogaster has expanded in recent years, but it still remains a young field with much to be done. The key benefit in using Drosophila as a model system for ethanol lies in the ease of genetic manipulation and the speed at which genetic modifications can be linked to behavioral phenotypes. It is a good "scouting organism" to identify candidate genes that can later be tested in more complex systems. In the first specific aim, we will conduct a large scale genetic screen to identify mutations that perturb rapid tolerance to ethanol in Drosophila. This is the first ethanol screen in Drosophila to look specifically for tolerance. Since the behavior being measured is slightly different than methods previously used, we are likely to see new results. We are using two approaches. The first is to systematically screen the entire third chromosome using large chromosomal deletions, then narrow down those results to single genes. The second approach is to screen a collection of candidate genes that have been identified by ethanol work in other model systems. Using both approaches gives us a wide net to identify genes involved in tolerance. The second specific aim is to determine the temporal gene expression patterns following ethanol exposure. mRNA will be purified from fly heads and the genes identified in aim 1 will be assayed for ethanol induced changes. For this aim, I will learn and use real-time RT-PCR. This will provide a substantial training experience that will aid in my development as a well rounded scientist. The third aim is to determine whether mutations that eliminate ethanol tolerance also influence ethanol preference behavior. The mutants identified in aim 1 will be tested, and various courses of ethanol treatment will be used to see the effects of both genotype and environment on preference to ethanol. Flies have a natural preference for ethanol, due to its presence in rotting fruit, so they are an ideal model system with which to conduct these tests. Completing these aims will not only add immediately to the ethanol research field (by identifying new genes involved in ethanol tolerance), it will add new insight into the genetics of the complex disease of alcoholism. It will expand the tools in place for the use of Drosophila as an ethanol model system. By studying how the genes of fruit flies affect their response to alcohol, we will gain an understanding of alcohol's effects that helps treatment and prevention of alcoholism in the future.

描述（由申请人提供）：尽管有广泛而多样的文献，但在乙醇研究领域仍有许多未解之谜。近年来，对乙醇对黑腹果蝇的影响的研究已经扩大，但它仍然是一个年轻的领域，有很多工作要做。使用果蝇作为乙醇模型系统的关键好处在于易于进行基因操作，并且可以快速地将遗传修饰与行为表型联系起来。它是一个很好的“侦察有机体”，可以识别候选基因，然后在更复杂的系统中进行测试。在第一个具体目标中，我们将进行大规模的基因筛选，以确定干扰果蝇对乙醇快速耐受性的突变。这是第一个在果蝇中专门寻找耐受性的乙醇筛选。由于所测量的行为与以前使用的方法略有不同，我们可能会看到新的结果。我们采用了两种方法。第一种方法是系统地筛选整个第三条染色体，使用大的染色体缺失，然后将结果缩小到单个基因。第二种方法是筛选一组候选基因，这些基因已经在其他模型系统中被乙醇工作识别出来。使用这两种方法为我们提供了一个广泛的网络来识别与耐受性有关的基因。第二个具体目标是确定乙醇暴露后的时间基因表达模式。将从蝇头中纯化mRNA，并检测目的1中鉴定的基因在乙醇诱导下的变化。为此，我将学习和使用实时RT-PCR。这将提供丰富的训练经验，这将有助于我发展成为一个全面发展的科学家。第三个目的是确定消除乙醇耐受性的突变是否也会影响乙醇偏好行为。将对目标1中确定的突变体进行测试，并使用不同的乙醇处理过程来观察基因型和环境对乙醇偏好的影响。苍蝇对乙醇有天然的偏好，因为它存在于腐烂的水果中，所以它们是进行这些测试的理想模型系统。完成这些目标不仅会立即增加乙醇研究领域（通过识别涉及乙醇耐受性的新基因），还会为复杂的酒精中毒疾病的遗传学提供新的见解。它将扩展现有的工具，将果蝇用作乙醇模型系统。通过研究果蝇的基因如何影响它们对酒精的反应，我们将了解酒精的作用，从而在未来帮助治疗和预防酒精中毒。