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A high-throughput fluorescence-based assay system for appetite-regulating gene and drug screening
Yasuhito Shimada1, Minoru Hirano, Yuhei Nishimura
1Department of Molecular and Cellular Pharmacology, Mie University Graduate School of Medicine, Tsu, Mie, Japan.
Plos One
|January 10, 2013
Summary
A new zebrafish assay enables rapid screening of appetite suppressants and enhancers. This method aids in discovering new obesity treatments by analyzing feeding behavior and gene expression similar to mammals.
Area of Science:
- Metabolic research
- Obesity and metabolic syndrome
- Pharmacological screening
Background:
- Rising global obesity and metabolic syndrome rates necessitate novel treatments.
- Current obesity pharmacotherapies are limited, primarily consisting of appetite suppressants and fat blockers.
- Lack of efficient screening methods hinders the discovery of new appetite-modulating agents.
Purpose of the Study:
- To develop a high-throughput assay for measuring feeding volume in zebrafish (Danio rerio).
- To validate the utility of this assay for screening appetite suppressants and enhancers.
- To investigate conserved appetite-regulating mechanisms between zebrafish and mammals.
Main Methods:
- Development of a rapid feeding volume measurement assay using fluorescently-labeled paramecia in zebrafish.
- Utilized a 96-well microplate reader for quantitative feeding volume assessment.
- Conducted gene expression analysis (bdnf), gene knockdown (NPY, Ins, MC4R, AgRP, CNR1), and administration of clinical appetite suppressants (fluoxetine, sibutramine, mazindol, phentermine, rimonabant).
- Integrated behavioral analysis to assess treatment-related adverse effects on locomotor activity.
Main Results:
- The zebrafish feeding assay demonstrated high throughput and reproducibility.
- Gene expression and knockdown studies revealed conserved appetite regulation pathways between zebrafish and mammals.
- Administration of known appetite suppressants in zebrafish mirrored their effects in mammals.
- Behavioral analysis identified potential adverse effects on locomotor activity.
Conclusions:
- A novel, rapid, and high-throughput zebrafish assay for measuring feeding volume has been successfully developed.
- This assay is suitable for screening potential appetite suppressants and enhancers.
- The study validates zebrafish as a model for investigating appetite regulation and discovering novel therapeutic targets for obesity.
