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Journal of Biomolecular Screening
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Article

A Whole-Animal Microplate Assay for Metabolic Rate Using Zebrafish

Khadijah Makky, Petar Duvnjak, Kallal Pramanik, Ramani Ramchandran, and Alan N. Mayer*

* To whom correspondence should be addressed. E-mail: alanmayer{at}mac.com.


  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 (npofW07-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. (Journal of Biomolecular Screening XXXX;xx-xx)

First published on November 21, 2008, doi:10.1177/1087057108326080

Journal of Biomolecular Screening 2008;13:960.

A more recent version of this article appeared on December 1, 2008

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