title: Luciferase reporter gene assay on human 5-HT receptor: which response element should be chosen?
abstract: 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.
authors.affiliation: Shanghai Key Laboratory of new drug design, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China.
authors.forename: Yiming
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Shanghai Key Laboratory of new drug design, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China.
authors.forename: Zhongyu
authors.initials: Z
authors.lastname: Xu
authors.affiliation: Shanghai Key Laboratory of new drug design, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China.
authors.forename: Dang
authors.initials: D
authors.lastname: Wu
authors.affiliation: Shanghai Key Laboratory of new drug design, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Li
authors.affiliation: School of Material Science and Engineering, Nanchang Hangkong University, Nanchang 330063, China.
authors.forename: Cheng
authors.initials: C
authors.lastname: Song
authors.affiliation: 1] Shanghai Key Laboratory of new drug design, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China [2] Shanghai Key Laboratory of Regulatory Biology, The Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai 200241, China.
authors.forename: Weiqiang
authors.initials: W
authors.lastname: Lu
authors.affiliation: Shanghai Key Laboratory of new drug design, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China.
authors.forename: Jin
authors.initials: J
authors.lastname: Huang
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D017930:Genes, Reporter; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism*; D011985:Receptors, Serotonin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020218:Response Elements / Q000235:genetics; D015398:Signal Transduction
chemical_list: D011985:Receptors, Serotonin; D008156:Luciferases