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Luciferase Reporter Gene Assay on Human 5-HT Receptor: Which Response Element Should Be Chosen?
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, bas...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4306921/ https://www.ncbi.nlm.nih.gov/pubmed/25622827 http://dx.doi.org/10.1038/srep08060 |
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author | Chen, Yiming Xu, Zhongyu Wu, Dang Li, Jian Song, Cheng Lu, Weiqiang Huang, Jin |
author_facet | Chen, Yiming Xu, Zhongyu Wu, Dang Li, Jian Song, Cheng Lu, Weiqiang Huang, Jin |
author_sort | Chen, Yiming |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-4306921 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-43069212015-02-06 Luciferase Reporter Gene Assay on Human 5-HT Receptor: Which Response Element Should Be Chosen? Chen, Yiming Xu, Zhongyu Wu, Dang Li, Jian Song, Cheng Lu, Weiqiang Huang, Jin Sci Rep Article 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. Nature Publishing Group 2015-01-27 /pmc/articles/PMC4306921/ /pubmed/25622827 http://dx.doi.org/10.1038/srep08060 Text en Copyright © 2015, Macmillan Publishers Limited. All rights reserved http://creativecommons.org/licenses/by-nc-nd/4.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder in order to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/4.0/ |
spellingShingle | Article Chen, Yiming Xu, Zhongyu Wu, Dang Li, Jian Song, Cheng Lu, Weiqiang Huang, Jin Luciferase Reporter Gene Assay on Human 5-HT Receptor: Which Response Element Should Be Chosen? |
title | Luciferase Reporter Gene Assay on Human 5-HT Receptor: Which Response Element Should Be Chosen? |
title_full | Luciferase Reporter Gene Assay on Human 5-HT Receptor: Which Response Element Should Be Chosen? |
title_fullStr | Luciferase Reporter Gene Assay on Human 5-HT Receptor: Which Response Element Should Be Chosen? |
title_full_unstemmed | Luciferase Reporter Gene Assay on Human 5-HT Receptor: Which Response Element Should Be Chosen? |
title_short | Luciferase Reporter Gene Assay on Human 5-HT Receptor: Which Response Element Should Be Chosen? |
title_sort | luciferase reporter gene assay on human 5-ht receptor: which response element should be chosen? |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4306921/ https://www.ncbi.nlm.nih.gov/pubmed/25622827 http://dx.doi.org/10.1038/srep08060 |
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