Identification of microRNAs that mediate ethanol behavioral responses

介导乙醇行为反应的 microRNA 的鉴定

基本信息

批准号：
9320642
负责人：
NIGEL S ATKINSON
金额：
$ 22.6万
依托单位：
UNIVERSITY OF TEXAS AT AUSTIN
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-08-01 至 2019-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9320642
关键词：
Acute Address Affect Alcohol consumption Alcohol dependence Alcohol withdrawal syndrome Alcoholism Alcohols Behavior Behavioral Biological Biological Assay Brain Calcium-Activated Potassium Channel Characteristics Collaborations Complex Data Dependence Development Disease Drosophila genus Epigenetic Process Ethanol Gene Expression Gene Rearrangement Genes Genetic Genetic Transcription Goals Health Individual Lead Mammals Mediating Messenger RNA MicroRNAs Modeling Molecular Nervous system structure Neuraxis Neurons Organ Pharmaceutical Preparations Phenotype Play Porifera Post-Transcriptional Regulation Production Public Health RNA Splicing Regulation Research Resistance Review Literature Role Sedation procedure Shapes System Testing Time Tissues Transcript Transgenes Translations United States Untranslated RNA Variant Withdrawal Symptom Work alcohol behavior alcohol exposure alcohol related problem alcohol response base behavioral response behavioral tolerance cost fly large-conductance calcium-activated potassium channels neuroadaptation next generation sequencing novel overexpression problem drinker public health relevance stem transcriptome transcriptome sequencing

项目摘要

DESCRIPTION (provided by applicant): At the core of the behavioral changes characteristic of alcoholism is the rearrangement of gene expression in the brain of an addicted individual. However, there exists a fundamental gap in our understanding of how alcohol consumption produces this rearrangement and leads to subsequent changes in behavior. Previous work in a mammalian system described how a particular microRNA (miRNA) acts as a key intermediary in the production of alcohol-induced changes in gene expression that produce functional-alcohol tolerance. The objective of this application is to combine the power of next generation sequencing with Drosophila genetics to identify all alcohol-sensitive miRNAs and to functionally test them for a role in producing an alcoholism-related behavior: functional ethanol tolerance (inducible ethanol resistance). This research goal stems from collaboration between two labs, one specializing in alcohol regulation of miRNA in the mammalian system, the other being an expert in fruit fly genetics and epigenetic mechanisms of alcohol actions. Based upon preliminary data and review of the literature, the central hypothesis is that even acute ethanol exposure causes long lasting changes in miRNA expression that lead to a rearrangement of the brain transcriptome. To test this hypothesis, miRNA-Seq and RNA-Seq will be performed on flies that have been briefly sedated with ethanol. Three different time points will be assessed in order to capture early and late changes in miRNA expression and their targets. Then, each ethanol-responsive miRNA will be functionally tested to determine if it contributes to functional ethanol tolerance (inducible-ethanol resistance). In flies, ethanol tolerance is a product only of neural adaptation to ethanol. Therefore, we will manipulate miRNA expression levels in the nervous system using the Gal4/UAS system and then determine its effect on functional ethanol tolerance and ethanol resistance. Achieving these goals will help identify and evaluate the role of miRNAs in the neural adaptations that underlie ethanol tolerance and will contribute to our longer-term goal to understand how ethanol consumption modulates gene expression to promote alcoholism.

描述（由适用提供）：酒精中毒的行为变化特征的核心是上瘾个体大脑中基因表达的重排。但是，我们对饮酒如何产生这种重排的理解存在根本差距，并导致行为变化。哺乳动物系统中的先前工作描述了特定的microRNA（miRNA）如何充当酒精诱导的基因表达变化的关键中间体，从而产生功能性杯子耐受性。该应用的目的是将下一代测序的力量与果蝇遗传学相结合，以鉴定所有对酒精敏感的miRNA，并在功能上测试它们在产生与酒精中毒相关的行为中的作用：功能性乙醇耐受性（诱导乙醇耐药性）。该研究目标源于两个实验室之间的合作，一个实验室在哺乳动物系统中专门从事miRNA的酒精调节，另一个是果蝇遗传学和酒精作用表观遗传机制的专家。基于初步数据和文献综述，中心假设是，即使急性乙醇暴露也会导致miRNA表达的持久变化，从而导致大脑转录组重新排列。为了检验这一假设，将对用乙醇短暂镇静的果蝇进行miRNA-Seq和RNA-Seq。将评估三个不同的时间点，以捕获miRNA表达及其目标的早期和晚期变化。然后，将对每个乙醇反应性miRNA进行功能测试，以确定它是否有助于功能性乙醇耐受性（诱导乙醇抗性）。在苍蝇中，乙醇耐受性仅是对乙醇的神经适应性的产物。因此，我们将使用GAL4/UAS系统操纵神经系统中的miRNA表达水平，然后确定其对功能性乙醇耐受性和乙醇耐药性的影响。实现这些目标将有助于识别和评估miRNA在乙醇耐受性的神经适应性中的作用，并将有助于我们的长期目标，以了解乙醇消耗如何调节基因表达以促进酒精中毒。