Probing the circadian clock in zebrafish larvae with in situ single cell transcri

用原位单细胞转录探测斑马鱼幼虫的生物钟

基本信息

批准号：
8787930
负责人：
Long Cai
金额：
$ 20.81万
依托单位：
CALIFORNIA INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-07-01 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8787930
关键词：
Abnormal coordination Acute Address Affect Animal Model Animals Behavior Behavior Disorders Behavioral Biological Assay Biological Models Bipolar Disorder Brain region Cells Chronic Circadian Rhythm Sleep Disorders Circadian Rhythms Complex Cues Defect Disease Drug Addiction Drug Modulation Drug abuse Dyes Ensure Environment Fluorescence Microscopy Fluorescent in Situ Hybridization Food Gene Expression Gene Expression Profiling Genes High Prevalence Hour Human In Situ Individual Label Larva Lead Light Link Major Depressive Disorder Measures Messenger RNA Methods Molecular Monitor Mood Disorders Oligonucleotides Organism Pattern Pharmaceutical Preparations Phase Physiological Processes Play Recovery Regulation Relative (related person)Resolution Rewards Role Scheme Seasonal Affective Disorder Sleep Techniques Technology Temperature Testing Therapeutic Time Tissues Transcript Vertebrates Zebrafish addiction base cell type circadian pacemaker combat fluorophore in vivo mRNA Expression multiplex detection new technology novel novel strategies novel therapeutic intervention public health relevance scale up single molecule small molecule tool

项目摘要

DESCRIPTION (provided by applicant): Disruption of normal sleep and circadian rhythms is a major factor affecting drug addiction and recovery. It has recently been shown that essentially all cells in our body have a circadian clock. Using zebrafish as a model system, we will apply and optimize a new technology in single cell gene expression analysis to characterize the diversity of behaviors in circadian clocks amongst cells in an organism. In particular, we will apply a method to profile mRNA expression levels in situ to measure the abundance of 25 circadian clock genes simultaneously in each cell of intact zebrafish larvae. This will provide a global assessment of circadian rhythms in all cells of an intact vertebrate animal at an unprecedented level of detail and robustness. We will test several hypotheses regarding circadian clock gene expression in vivo and its modulation by drugs associated with addiction and reward. We will also test the hypothesis that modulating circadian rhythms affects behaviors associated with reward and addiction. These studies could reveal novel circadian mechanisms that affect drug abuse and addiction and may lead to new therapies for these disorders. If successful, this technology can be extended to other model organisms to study large-scale gene expression changes associated with many complex behaviors.

描述（由申请人提供）：正常睡眠和昼夜节律的破坏是影响药物成瘾和恢复的主要因素。最近已经显示，本质上，我们体内的所有细胞都有昼夜节律。使用斑马鱼作为模型系统，我们将应用和优化单细胞基因表达分析中的新技术，以表征生物体中细胞中昼夜节律时钟中行为的多样性。特别是，我们将在原位介绍mRNA表达水平，以同时在完整斑马鱼幼虫的每个细胞中同时测量25个昼夜节律基因的丰度。这将在完整的脊椎动物动物的所有细胞中以前所未有的细节和鲁棒性水平提供对昼夜节律的全球评估。我们将检验几个关于体内昼夜节律时钟基因表达的假设及其对成瘾和奖励相关的药物的调节。我们还将检验以下假设：调节昼夜节律会影响与奖励和成瘾有关的行为。这些研究可能揭示了影响药物滥用和成瘾的新型昼夜节律机制，并可能导致这些疾病的新疗法。如果成功，该技术可以扩展到其他模型生物体，以研究与许多复杂行为相关的大规模基因表达变化。