High throughput Danio rerio energy expenditure assay

Savannah Y. Williams, Benjamin J Renquist

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

Zebrafish are an important model organism with inherent advantages that have the potential to make zebrafish a widely applied model for the study of energy homeostasis and obesity. The small size of zebrafish allows for assays on embryos to be conducted in a 96- or 384-well plate format, Morpholino and CRISPR based technologies promote ease of genetic manipulation, and drug treatment by bath application is viable. Moreover, zebrafish are ideal for forward genetic screens allowing for novel gene discovery. Given the relative novelty of zebrafish as a model for obesity, it is necessary to develop tools that fully exploit these benefits. Herein, we describe a method to measure energy expenditure in thousands of embryonic zebrafish simultaneously. We have developed a whole animal microplate platform in which we use 96-well plates to isolate individual fish and we assess cumulative NADH2 production using the commercially available cell culture viability reagent alamarBlue. In poikilotherms the relationship between NADH2 production and energy expenditure is tightly linked. This energy expenditure assay creates the potential to rapidly screen pharmacological or genetic manipulations that directly alter energy expenditure or alter the response to an applied drug (e.g. insulin sensitizers).

Original languageEnglish (US)
Article numbere53297
JournalJournal of Visualized Experiments
Volume2016
Issue number107
DOIs
Publication statusPublished - Jan 27 2016

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Keywords

  • Danio rerio
  • Drug discovery
  • Energy expenditure
  • High-throughput
  • Issue 107
  • Metabolic syndrome
  • Molecular biology
  • Obesity
  • Oxygen consumption
  • Zebrafish

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Immunology and Microbiology(all)
  • Biochemistry, Genetics and Molecular Biology(all)

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