Department of Molecular and Cellular Pharmacology, Mie University Graduate School of Medicine, Tsu, Mie, Japan.
PLoS One. 2012;7(12):e52549. doi: 10.1371/journal.pone.0052549. Epub 2012 Dec 26.
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.
越来越多的人患有代谢综合征和肥胖症，这不仅是发达国家，也是发展中国家面临的一个严重问题。然而，目前用于治疗肥胖症的药物很少。现有的药物主要是食欲抑制剂和胃肠道脂肪阻滞剂。我们开发了一种简单快速的斑马鱼（Danio rerio）摄食量测量方法。该方法可用于筛选食欲抑制剂和增强剂。在这项研究中，我们用荧光标记的活草履虫喂养斑马鱼，并使用 96 孔微孔板读数仪测量摄食量。通过脑源性神经营养因子（bdnf）的基因表达分析、食欲调节基因（神经肽 Y、前胰岛素原、黑素皮质素 4 受体、刺鼠相关蛋白和大麻素受体 1）的敲低以及临床食欲抑制剂（氟西汀、西布曲明、mazindol、苯丙醇胺和利莫那班）的给药，揭示了斑马鱼和哺乳动物中调节食欲的机制的相似性。结合行为分析，我们能够评估基因敲低和化学处理对运动活动的不良影响。总之，我们开发了一种使用斑马鱼的测定方法，可用于食欲抑制剂和增强剂的高通量筛选和靶基因发现。
