PUBLICATION
A Whole-Animal Microplate Assay for Metabolic Rate Using Zebrafish
- Authors
- Makky, K., Duvnjak, P., Pramanik, K., Ramchandran, R., and Mayer, A.N.
- ID
- ZDB-PUB-081203-9
- Date
- 2008
- Source
- Journal of Biomolecular Screening 13(10): 960-967 (Journal)
- Registered Authors
- Mayer, Alan, Ramchandran, Ramani
- Keywords
- zebrafish, energy metabolism, genetic screening, chemical genetics, drug screening, energy expenditure, metabolic regulation
- MeSH Terms
-
- 2,4-Dinitrophenol/pharmacology
- Acids
- Aging/drug effects
- Animals
- Biological Assay/methods*
- Energy Metabolism*/drug effects
- Energy Metabolism*/genetics
- Genotype
- Larva/drug effects
- Larva/metabolism
- Phenotype
- Sirolimus/pharmacology
- Temperature
- Zebrafish/metabolism*
- PubMed
- 19029015 Full text @ J. Biomol. Screen.
Citation
Makky, K., Duvnjak, P., Pramanik, K., Ramchandran, R., and Mayer, A.N. (2008) A Whole-Animal Microplate Assay for Metabolic Rate Using Zebrafish. Journal of Biomolecular Screening. 13(10):960-967.
Abstract
Regulation of whole-body metabolism and energy homeostasis has been shown to require signaling between multiple organs. To identify genetic programs that determine metabolic rate, and compounds that can modify it, a whole-animal assay amenable to large-scale screening was developed. The direct correlation of acid production with metabolic rate was exploited to use a noninvasive colorimetric assay for acid secretion by individual zebrafish larvae in a 96-well plate format. A 3-fold increase in metabolic rate was detected that accompanied development between 24 and 96 h postfertilization. Dynamic changes in metabolic rate were also detected in response to different conditions such as temperature and drug treatments, in general agreement with the rate of oxygen consumption measured concomitantly. This assay was used to measure metabolic rate in the progeny of fish known to carry a recessive mutation in a gene required for ribosome biogenesis (npo(fW07-g)), which would be expected to reduce energy consumption. A strong correlation was found (p < 10(-6)) between reduced metabolic rate and genotype even before the developmental defect was visually evident. These studies support the conclusion that whole-animal acid secretion can be used as a readout for energy metabolism, thus enabling large-scale screening for genetic and chemical regulators of metabolic rate in a vertebrate.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping