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2013/01/04
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A High-Throughput Fluorescence-Based Assay System for Appetite-Regulating Gene and Drug Screening

Abstract
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.

Citation: Shimada Y, Hirano M, Nishimura Y, Tanaka T (2012) A High-Throughput Fluorescence-Based Assay System for Appetite-Regulating Gene and Drug Screening. PLoS ONE 7(12): e52549. doi:10.1371/journal.pone.0052549

Editor: Zhiyuan Gong, National University of Singapore, Singapore

Received: August 13, 2012; Accepted: November 20, 2012; Published: December 26, 2012

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