Monitoring DAT in Live Rats

监测活鼠 DAT

• 批准号: 8440730
• 负责人: Zhicheng Carl Lin
• 金额: $ 18.96万
• 依托单位: MCLEAN HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-03-15 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8440730
• 关键词: Address; Affect; Animals; Attention; Behavior; Behavioral; Bioinformatics; Bipolar Disorder; Brain; Brain imaging; Cell Surface Proteins; Cerebrospinal Fluid; Cocaine; Cognition; Communities; Complement; Corpus striatum structure; DNA Sequence; Disease; Dopamine; Drug abuse; Environment; Enzymes; Etiology; Evaluation; Gene Targeting; Genes; Genetic; Genetic Engineering; Genomics; Half-Life; Human; In Vitro; Investigation; Knowledge; Left; Life; Liquid substance; Luciferases; Measures; Mental Depression; Messenger RNA; Microdialysis; Midbrain structure; Modeling; Monitor; Mood stabilizers; Motivation; Movement; Mus; Neurosciences; Pathologic Processes; Pharmaceutical Preparations; Phenotype; Physiological; Proteins; Rattus; Regulatory Element; Reporter; Reporter Genes; Reporting; Risk; Risk Factors; Rodent; Short-Term Memory; Substance abuse problem; Synaptosomes; Technology; Therapeutic; Variant; Work; Zinc Fingers; abstracting; base; density; design; dopamine transporter; dopaminergic neuron; drug of abuse; fusion gene; genetic association; in vivo; inhibitor/antagonist; meetings; mutant; neuropsychiatry; nuclease; psychostimulant; response; small molecule; smoking cessation; transmission process; uptake; valproate

DESCRIPTION (provided by applicant): Monitoring DAT in Live Mice Abstract Expressed exclusively in dopaminergic neurons, the dopamine transporter gene (DAT/SLC6A3) is a risk factor for many diseases including drug abuse, depression and bipolar disorder, based on hundreds of association studies. Since DNA sequence variation in the human DAT is correlated with striatal DAT protein levels in humans, it is postulated that altered DAT activity confers the risk. However, it is not known whether or not altered DAT activity is indeed correlated with abnormal behaviors or diseases. To fill in this critical knowledge gap, here we propose to develop a mouse line where DAT carries a reporter gene that encodes a secreted Gaussia luciferase (GLuc) with desired reporter qualities such as short in vivo half-life, high sensitivity and large dynamic range. This DAT-GLuc line will allow monitoring DAT activity sequentially in the same live mice under various environments including drugs of abuse, simply by measuring GLuc activity in the circulated fluid. DAT-GLuc aims to follow the DAT activity and correlate the activity with behaviors or efficacy of DAT-related medications in mice. This gene activity-reporting mouse line will complement the current in vivo technologies including brain imaging (for proteins) and microdialysis (for small molecules).

描述（由申请人提供）：监测活体小鼠中的DAT摘要多巴胺转运蛋白基因（DAT/SLC 6A 3）仅在多巴胺能神经元中表达，是许多疾病的危险因素，包括药物滥用、抑郁症和双相情感障碍，基于数百项相关研究。由于人类DAT中的DNA序列变异与人类纹状体DAT蛋白水平相关，因此推测DAT活性改变会带来风险。然而，目前尚不清楚DAT活性的改变是否确实与异常行为或疾病相关。为了填补这一关键的知识空白，在这里，我们建议开发一种小鼠品系，其中DAT携带报告基因，该报告基因编码具有所需报告品质的分泌型高斯荧光素酶（GLuc），例如短的体内半衰期，高灵敏度 和大的动态范围。该DAT-GLuc线将允许在包括滥用药物的各种环境下连续监测相同活小鼠中的DAT活性，简单地通过测量循环流体中的GLuc活性。DAT-GLuc旨在跟踪DAT活性，并将该活性与小鼠中DAT相关药物的行为或疗效相关联。这种基因活性报告小鼠系将补充目前的体内技术，包括脑成像（蛋白质）和微透析（小分子）。