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A High-Throughput Fluorescence-Based Assay System for Appetite-Regulating Gene and Drug Screening
The increasing number of people suffering from metabolic syndrome and obesity is becoming a serious problem not only in developed countries, but also in developing countries. However, there are few agents currently approved for the treatment of obesity. Those that are available are mainly appetite s...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3530442/ https://www.ncbi.nlm.nih.gov/pubmed/23300705 http://dx.doi.org/10.1371/journal.pone.0052549 |
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author | Shimada, Yasuhito Hirano, Minoru Nishimura, Yuhei Tanaka, Toshio |
author_facet | Shimada, Yasuhito Hirano, Minoru Nishimura, Yuhei Tanaka, Toshio |
author_sort | Shimada, Yasuhito |
collection | PubMed |
description | The increasing number of people suffering from metabolic syndrome and obesity is becoming a serious problem not only in developed countries, but also in developing countries. However, there are few agents currently approved for the treatment of obesity. Those that are available are mainly appetite suppressants and gastrointestinal fat blockers. We have developed a simple and rapid method for the measurement of the feeding volume of Danio rerio (zebrafish). This assay can be used to screen appetite suppressants and enhancers. In this study, zebrafish were fed viable paramecia that were fluorescently-labeled, and feeding volume was measured using a 96-well microplate reader. Gene expression analysis of brain-derived neurotrophic factor (bdnf), knockdown of appetite-regulating genes (neuropeptide Y, preproinsulin, melanocortin 4 receptor, agouti related protein, and cannabinoid receptor 1), and the administration of clinical appetite suppressants (fluoxetine, sibutramine, mazindol, phentermine, and rimonabant) revealed the similarity among mechanisms regulating appetite in zebrafish and mammals. In combination with behavioral analysis, we were able to evaluate adverse effects on locomotor activities from gene knockdown and chemical treatments. In conclusion, we have developed an assay that uses zebrafish, which can be applied to high-throughput screening and target gene discovery for appetite suppressants and enhancers. |
format | Online Article Text |
id | pubmed-3530442 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-35304422013-01-08 A High-Throughput Fluorescence-Based Assay System for Appetite-Regulating Gene and Drug Screening Shimada, Yasuhito Hirano, Minoru Nishimura, Yuhei Tanaka, Toshio PLoS One Research Article The increasing number of people suffering from metabolic syndrome and obesity is becoming a serious problem not only in developed countries, but also in developing countries. However, there are few agents currently approved for the treatment of obesity. Those that are available are mainly appetite suppressants and gastrointestinal fat blockers. We have developed a simple and rapid method for the measurement of the feeding volume of Danio rerio (zebrafish). This assay can be used to screen appetite suppressants and enhancers. In this study, zebrafish were fed viable paramecia that were fluorescently-labeled, and feeding volume was measured using a 96-well microplate reader. Gene expression analysis of brain-derived neurotrophic factor (bdnf), knockdown of appetite-regulating genes (neuropeptide Y, preproinsulin, melanocortin 4 receptor, agouti related protein, and cannabinoid receptor 1), and the administration of clinical appetite suppressants (fluoxetine, sibutramine, mazindol, phentermine, and rimonabant) revealed the similarity among mechanisms regulating appetite in zebrafish and mammals. In combination with behavioral analysis, we were able to evaluate adverse effects on locomotor activities from gene knockdown and chemical treatments. In conclusion, we have developed an assay that uses zebrafish, which can be applied to high-throughput screening and target gene discovery for appetite suppressants and enhancers. Public Library of Science 2012-12-26 /pmc/articles/PMC3530442/ /pubmed/23300705 http://dx.doi.org/10.1371/journal.pone.0052549 Text en © 2012 Shimada et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Shimada, Yasuhito Hirano, Minoru Nishimura, Yuhei Tanaka, Toshio A High-Throughput Fluorescence-Based Assay System for Appetite-Regulating Gene and Drug Screening |
title | A High-Throughput Fluorescence-Based Assay System for Appetite-Regulating Gene and Drug Screening |
title_full | A High-Throughput Fluorescence-Based Assay System for Appetite-Regulating Gene and Drug Screening |
title_fullStr | A High-Throughput Fluorescence-Based Assay System for Appetite-Regulating Gene and Drug Screening |
title_full_unstemmed | A High-Throughput Fluorescence-Based Assay System for Appetite-Regulating Gene and Drug Screening |
title_short | A High-Throughput Fluorescence-Based Assay System for Appetite-Regulating Gene and Drug Screening |
title_sort | high-throughput fluorescence-based assay system for appetite-regulating gene and drug screening |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3530442/ https://www.ncbi.nlm.nih.gov/pubmed/23300705 http://dx.doi.org/10.1371/journal.pone.0052549 |
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