文献类型：期刊文献

英文题名：Luciferase Reporter Gene Assay on Human 5-HT Receptor: Which Response Element Should Be Chosen?

作者：Chen, Yiming[1];Xu, Zhongyu[1];Wu, Dang[1];Li, Jian[1];Song, Cheng[2];Lu, Weiqiang[1,3,4];Huang, Jin[1]

机构：[1]E China Univ Sci & Technol, Sch Pharm, Shanghai Key Lab New Drug Design, Shanghai 200237, Peoples R China;[2]Nanchang Hangkong Univ, Sch Mat Sci & Engn, Nanchang 330063, Peoples R China;[3]E China Normal Univ, Inst Biomed Sci, Shanghai Key Lab Regulatory Biol, Shanghai 200241, Peoples R China;[4]E China Normal Univ, Sch Life Sci, Shanghai 200241, Peoples R China

年份：2015

卷号：5

外文期刊名：SCIENTIFIC REPORTS

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000348351800010)】；

基金：This work was supported by the National Natural Science Foundation of China (Grants 81402482, 91313303, 81102420 and 51003044), the Shanghai Committee of Science and Technology (14ZR1411100), China Postdoctoral Science Foundation grant (2014M551361), the Jiangxi Natural Science Foundation (20132BAB206035) and the Floor plan of scientific and technological projects in Jiangxi (KJLD13023).

语种：英文

摘要：Serotonin (5-HT) receptors are valuable molecular targets for antipsychotic drug discovery. Current reported methods for detecting 5-HT receptors, such as cAMP accumulation and calcium influx assay, are often demanding specialized instruments and inconvenient. The luciferase reporter gene assay, based on the responsible-element-regulated expression of luciferase, has been widely applied in the high-throughput functional assay for many targets because of its high sensitivity and reliability. However, 5-HT receptors couple to multiple G-proteins regulate respective downstream signalling pathways and are usually detected using different response elements. Hence, finding a suitable response element to fulfil the detection of different 5-HT receptors and make the results of luciferase reporter gene assays generalizable is very useful for active compounds screening. Here, we conducted three luciferase reporter assays using CRE, NFAT, and SRE response elements attached to 5-HT to detect the activation of different 5-HT receptors in CHO-K1 cells. The potencies and efficacies of the reported ligands (agonists and antagonists) were determined and compared. Our results indicate that CRE-luciferase reporter gene is sensitive and reliable to detect the activities of G protein-coupled 5-HT receptors.