PUBLICATION

High Throughput Danio Rerio Energy Expenditure Assay

Authors
Williams, S.Y., Renquist, B.J.
ID
ZDB-PUB-160211-1
Date
2016
Source
Journal of visualized experiments : JoVE   (107): e53297 (Journal)
Registered Authors
Williams, Savannah
Keywords
Molecular Biology, Energy Expenditure, Oxygen Consumption, Danio rerio, Zebrafish, Drug Discovery, High-throughput, Obesity, Metabolic Syndrome
MeSH Terms
  • Animals
  • Biological Assay
  • Energy Metabolism/physiology*
  • Indicators and Reagents/pharmacokinetics
  • Morpholinos/pharmacokinetics*
  • Oxazines/pharmacokinetics
  • Xanthenes/pharmacokinetics
  • Zebrafish/embryology*
PubMed
26863590 Full text @ J. Vis. Exp.
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).
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping