High Throughput Danio Rerio Energy Expenditure Assay

Area of Science:

• * Comparative physiology and endocrinology.
• * Developmental biology and toxicology.
• * Genetics and genomics.

Background:

• * Zebrafish (Danio rerio) are increasingly recognized as a valuable model organism for studying energy homeostasis and obesity.
• * Existing advantages include small size, suitability for high-throughput screening, ease of genetic manipulation (Morpholino, CRISPR), and simple drug administration.
• * Novel tools are needed to fully leverage zebrafish for obesity research, particularly for measuring energy expenditure.

Purpose of the Study:

• * To describe a novel method for simultaneously measuring energy expenditure in thousands of embryonic zebrafish.
• * To establish a high-throughput screening platform for identifying genetic or pharmacological modulators of energy expenditure.

Main Methods:

• * Development of a whole-animal microplate platform using 96-well plates to house individual embryonic zebrafish.
• * Assessment of cumulative NADH2 production as a proxy for energy expenditure using the alamarBlue reagent.
• * Utilizing the established link between NADH2 production and energy expenditure in poikilothermic organisms.

Main Results:

• * Successful implementation of a microplate-based assay capable of measuring energy expenditure in thousands of zebrafish embryos concurrently.
• * Demonstration of the assay's utility for rapid screening of interventions affecting energy expenditure.

Conclusions:

• * The developed microplate assay provides a powerful tool for high-throughput screening in zebrafish obesity research.
• * This method facilitates the rapid assessment of pharmacological and genetic manipulations impacting energy expenditure and drug responses (e.g., insulin sensitizers).
• * The assay enhances the utility of zebrafish as a model for understanding energy balance and developing obesity treatments.