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High-throughput assay for small molecules that modulate zebrafish embryonic heart rate

Nature Chemical Biology volume 1, pages 263–264 (2005)Cite this article

Abstract

To increase the facility and throughput of scoring phenotypic traits in embryonic zebrafish, we developed an automated micro-well assay for heart rate using automated fluorescence microscopy of transgenic embryos expressing green fluorescent protein in myocardium. The assay measures heart rates efficiently and accurately over a large linear dynamic range, and it rapidly characterizes dose dependence and kinetics of small molecule–induced changes in heart rate. This is the first high-throughput micro-well assay for organ function in an intact vertebrate.

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Figure 1: High-throughput heart-rate assay.
Figure 2: Rapid characterization of dose dependence and kinetics of response to drugs that decrease heart rate.

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Acknowledgements

We thank C. Felts for assistance with the automated microscope and Metamorph (Molecular Devices) software, D. Gilmour for providing the GFP expression plasmid pG1 and R. Peterson for use of the automated microscope. This work was funded by the US National Institutes of Health (NIH) grant 5T32HL07208-26 to Massachusetts General Hospital. C.G.B. was funded by National Institutes on Aging/NIH grant 1K01AG023562-01 and National Institute of Neurological Disorders and Stroke/NIH grant 1R03NS050806-01.

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Author notes

  1. Eric J Grande, Wolfgang Rottbauer & Mark C Fishman

    Present address: Present addresses: Novartis Institutes for Biomedical Research Inc., 250 Massachusetts Avenue, Cambridge, Massachusetts, 02139, USA (E.J.G., M.C.F.) and Department of Medicine III, University of Heidelberg, D-69120, Heidelberg, Germany (W.R.).,

Authors and Affiliations

  1. Developmental Biology Laboratory, Cardiovascular Research Center, Massachusetts General Hospital, 149 13th Street, Charlestown, 02129, Massachusetts, USA

    C Geoffrey Burns, David J Milan, Eric J Grande, Wolfgang Rottbauer, Calum A MacRae & Mark C Fishman

  2. Department of Medicine, Harvard Medical School, Boston, 02115, Massachusetts, USA

    C Geoffrey Burns, David J Milan, Eric J Grande, Wolfgang Rottbauer, Calum A MacRae & Mark C Fishman

Authors
  1. C Geoffrey Burns
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  2. David J Milan
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  3. Eric J Grande
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Corresponding author

Correspondence to C Geoffrey Burns.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Pre-exposing wells to fluorescent light prior to subsite examination increased assay efficiency. (PDF 51 kb)

Supplementary Fig. 2 (PDF 77 kb)

Supplementary Table 1

Automated microscopy algorithm for acquisition and analysis of video images for measuring fluorescent heart rates of Tg(cmlc2:GFP) embryos arrayed in 96-well plates. (PDF 121 kb)

Supplementary Methods (PDF 35 kb)

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Burns, C., Milan, D., Grande, E. et al. High-throughput assay for small molecules that modulate zebrafish embryonic heart rate. Nat Chem Biol 1, 263–264 (2005). https://doi.org/10.1038/nchembio732

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