Monitoring DAT in Live Rat

摘要根据数百项相关研究，多巴胺转运蛋白基因（DAT/SLC6A3）仅在多巴胺能神经元中表达，是包括药物滥用、抑郁症和双相情感障碍在内的许多疾病的危险因素。由于人类DAT中的DNA序列变化与人类纹状体DAT蛋白水平相关，因此假设DAT活性的改变会带来风险。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，我们建议开发一种小鼠系，其中DAT携带一个报告基因，该基因编码分泌的高斯荧光素酶（GLuc），具有所需的报告品质，如体内半衰期短，灵敏度高

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). PUBLIC HEALTH RELEVANCE: Monitoring DAT in live mice Project Narrative This research will generate a mouse line that uses luciferase to report the activity of the dopamine transporter gene (DAT), whose human orthologue is a risk factor for more than ten neuropsychiatric diseases. This DAT-reporting line will provide the research community a new tool to model and treat DAT-related diseases.