Monitoring DAT in Live Rats

描述(申请人提供)：监测活体小鼠的DAT摘要基于数百项关联研究，多巴胺转运蛋白基因(DAT/SLC6A3)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。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活性的改变与这种风险有关。然而，目前尚不清楚DAT活性改变是否真的与异常行为或疾病有关。为了填补这一关键的知识空白，我们建议开发一种小鼠系，其中DAT携带报告基因，该报告基因编码具有所需的报告性质的Gaussia Luciferase(GLuc)，例如在体内的半衰期短，高灵敏度 动态范围大。这条DAT-GLuc线只需通过测量循环液中的GLuc活性，就可以在包括滥用药物在内的各种环境下连续监测同一活体小鼠的DAT活动。DAT-GLuc旨在跟踪DAT活性，并将该活性与DAT相关药物的行为或疗效相关联。这种报告基因活性的小鼠品系将补充目前的活体技术，包括脑成像(蛋白质)和微透析(小分子)。

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).