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Rapid behavior-based identification of neuroactive small molecules in the zebrafish

Nature Chemical Biology volume 6, pages 231–237 (2010)Cite this article

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Abstract

Neuroactive small molecules are indispensable tools for treating mental illnesses and dissecting nervous system function. However, it has been difficult to discover novel neuroactive drugs. Here, we describe a high-throughput, behavior-based approach to neuroactive small molecule discovery in the zebrafish. We used automated screening assays to evaluate thousands of chemical compounds and found that diverse classes of neuroactive molecules caused distinct patterns of behavior. These 'behavioral barcodes' can be used to rapidly identify new psychotropic chemicals and to predict their molecular targets. For example, we identified new acetylcholinesterase and monoamine oxidase inhibitors using phenotypic comparisons and computational techniques. By combining high-throughput screening technologies with behavioral phenotyping in vivo, behavior-based chemical screens can accelerate the pace of neuroactive drug discovery and provide small-molecule tools for understanding vertebrate behavior.

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Figure 1: The PMR and high-throughput behavioral barcoding.
Figure 2: Neuroactive chemicals cause specific patterns of behavior.
Figure 3: Hierarchical clustering reveals that compounds cluster with functionally similar molecules.
Figure 4: Behavior-based discovery of novel neuroactive small molecules.
Figure 5: Chemical suppression of behavioral phenotypes.

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Acknowledgements

We thank E. Scolnick, M. Granato, J. Dowling, D. Milan, C. Felts, J. Rihel, A. Schier and members of our research groups for encouragement and advice. This work was supported by US National Institutes of Health training grant HL07208 (D.K.) and grants NS063733 (R.T.P.), MH085205 (R.T.P.), MH086867 (R.T.P.) and GM71896 (B.K.S. and J. Irwin), the National Sciences and Engineering Council of Canada (J.B.), the Canadian Institutes of Health Research (J.B.), the Max Kade Foundation (C.L.) and the Stanley Medical Research Institute (S.J.H.).

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Authors and Affiliations

  1. Cardiovascular Research Center and Division of Cardiology, Department of Medicine, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts, USA

    David Kokel, Chung Yan J Cheung, Rita Mateus, David Healey, Sonia Kim, Andreas A Werdich, Calum A MacRae & Randall T Peterson

  2. Broad Institute, Cambridge, Massachusetts, USA

    David Kokel, Chung Yan J Cheung, Rita Mateus, David Healey, Sonia Kim, Stephen J Haggarty & Randall T Peterson

  3. Department of Statistics, University of British Columbia, Vancouver, British Columbia, Canada

    Jennifer Bryan & Rick White

  4. Michael Smith Laboratories, University of British Columbia, Vancouver, British Columbia, Canada

    Jennifer Bryan

  5. Department of Pharmaceutical Chemistry, University of California, San Francisco, California, USA

    Christian Laggner & Brian Shoichet

  6. Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, USA

    Stephen J Haggarty

  7. Stanley Center for Psychiatric Research, Cambridge, Massachusetts, USA

    Stephen J Haggarty

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  1. David Kokel
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Contributions

D.K. designed and performed the research, analyzed the data and wrote the manuscript. J.B., C.L., R.W. and B.S. analyzed and interpreted the data and contributed to the manuscript. C.Y.J.C., R.M., D.H. and S.K. performed experiments. A.A.W. contributed to hardware design. S.J.H. and C.A.M. contributed reagents. R.T.P designed the research, analyzed the data and wrote the manuscript. All authors contributed to data interpretation and commented on the manuscript.

Corresponding authors

Correspondence to David Kokel or Randall T Peterson.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–5, Supplementary Tables 1–2 and Supplementary Methods (PDF 1464 kb)

Supplementary Movie 1

Movie of the PMR in zebrafish embryos in a petri dish at low magnification. (MOV 262 kb)

Supplementary Movie 2

Movie of the PMR at higher magnification. (MOV 455 kb)

Supplementary Movie 3

Movie of the PMR behavior at 30 hpf, showing that animals do not normally respond to a second pulse of light. (MOV 365 kb)

Supplementary Movie 4

Movie of the robotic screening hardware delivering light pulses to the individual wells of a 96-well plate. (MOV 594 kb)

Supplementary Movie 5

Movie of an untreated control well in the ETR assay. (MOV 444 kb)

Supplementary Movie 6

Movie of the slow to relax (STR) phenotype in a well treated with STR-1 during the ETR assay. (MOV 447 kb)

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Kokel, D., Bryan, J., Laggner, C. et al. Rapid behavior-based identification of neuroactive small molecules in the zebrafish. Nat Chem Biol 6, 231–237 (2010). https://doi.org/10.1038/nchembio.307

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