Article | Published:

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

Nature Chemical Biology volume 6, pages 231237 (2010) | Download Citation

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.

  • Compound C11H17NO3

    isoproterenol

  • Compound C16H13ClN2O

    diazepam

  • Compound C17H17NO2

    apomorphine

  • Compound C23H34O4

    digitoxigenin

  • Compound C13H14N4O2

    6-nitroquipazine

  • Compound C12H18Cl2N2O

    Clenbuterol

  • Compound C9H13NO3

    Epinepherine

  • Compound C35H41N5O5

    Dihydroergocristine

  • Compound C17H17NO2

    Dihydrexidine

  • Compound C14H19NO3

    3,4,4a,10b-Tetrahydro-4-propyl-2H,5H-(1)benzopyrano(4,3-b)-1,4-oxazin-9-ol

  • Compound C34H47NO11

    aconitine

  • Compound C36H51NO11

    Veratridine

  • Compound C25H32O7

    Strophanthidinic acid

  • Compound C14H19N3O2

    Eserine

  • Compound C18H22N2O3

    2-(4-Azepan-1-yl-4-oxo-butyl)-isoindole-1,3-dione

  • Compound C18H9ClN4O

    10-(2-Chlorophenyl)-1,3,4,10a-tetraaza-cyclopenta[b]fluoren-9-one

  • Compound C20H16O4

    6-Oxo-7,8,9,10-tetrahydrobenzo[c]chromen-3-yl benzoate

  • Compound C11H13N

    Pargyline

  • Compound C19H16O4

    Warfarin

  • Compound C10H12N3O3PS2

    Azinphos methyl

  • Compound C17H23NO3

    Atropine

  • Compound C7H9N2O+

    Pralidoxime

  • Compound C23H28N2O3

    Bopindolol

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References

  1. 1.

    & Discoveries in Biological Psychiatry (Lippincott, Philadelphia, 1970).

  2. 2.

    et al. How can drug discovery for psychiatric disorders be improved? Nat. Rev. Drug Discov. 6, 189–201 (2007).

  3. 3.

    Chemical genetics: ligand-based discovery of gene function. Nat. Rev. Genet. 1, 116–125 (2000).

  4. 4.

    , , & Combination chemical genetics. Nat. Chem. Biol. 4, 674–681 (2008).

  5. 5.

    Cooperative activation of action potential Na+ ionophore by neurotoxins. Proc. Natl. Acad. Sci. USA 72, 1782–1786 (1975).

  6. 6.

    Membrane adenosinetriphosphatase: a digitalis receptor? Science 198, 569–574 (1977).

  7. 7.

    , , , & Quantifying the relationships among drug classes. J. Chem. Inf. Model. 48, 755–765 (2008).

  8. 8.

    et al. Relating protein pharmacology by ligand chemistry. Nat. Biotechnol. 25, 197–206 (2007).

  9. 9.

    et al. Inhibition of monoamine oxidases by functionalized coumarin derivatives: biological activities, QSARs, and 3D-QSARs. J. Med. Chem. 43, 4747–4758 (2000).

  10. 10.

    et al. Quantitative structure-activity relationship and complex network approach to monoamine oxidase A and B inhibitors. J. Med. Chem. 51, 6740–6751 (2008).

  11. 11.

    Organophosphates/nerve agent poisoning: mechanism of action, diagnosis, prophylaxis, and treatment. Adv. Clin. Chem. 38, 151–216 (2004).

  12. 12.

    et al. Physostigmine: improvement of long-term memory processes in normal humans. Science 201, 272–274 (1978).

  13. 13.

    , & MAOIs in the contemporary treatment of depression. Neuropsychopharmacology 12, 185–219 (1995).

  14. 14.

    et al. Rapid mutation of endogenous zebrafish genes using zinc finger nucleases made by Oligomerized Pool ENgineering (OPEN). PLoS ONE 4, e4348 (2009).

  15. 15.

    , , , & Targeted gene inactivation in zebrafish using engineered zinc-finger nucleases. Nat. Biotechnol. 26, 695–701 (2008).

  16. 16.

    et al. Heritable targeted gene disruption in zebrafish using designed zinc-finger nucleases. Nat. Biotechnol. 26, 702–708 (2008).

  17. 17.

    & Re-evaluation of animal numbers and costs for in vivo tests to accomplish REACH legislation requirements for chemicals — a report by the Transatlantic Think Tank for Toxicology (t4). ALTEX 26, 187–208 (2009).

  18. 18.

    & A peroxidase-coupled continuous absorbance plate-reader assay for flavin monoamine oxidases, copper-containing amine oxidases and related enzymes. Nat. Protocols 1, 2498 (2006).

  19. 19.

    , & Daylight theory manual (Daylight Chemical Information Systems, 1995).

  20. 20.

    Chemoinformatics in Drug Discovery. (Wiley, Weinheim, Germany, 2005).

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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.).

Author information

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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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.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to David Kokel or Randall T Peterson.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–5, Supplementary Tables 1–2 and Supplementary Methods

Videos

  1. 1.

    Supplementary Movie 1

    Movie of the PMR in zebrafish embryos in a petri dish at low magnification.

  2. 2.

    Supplementary Movie 2

    Movie of the PMR at higher magnification.

  3. 3.

    Supplementary Movie 3

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

  4. 4.

    Supplementary Movie 4

    Movie of the robotic screening hardware delivering light pulses to the individual wells of a 96-well plate.

  5. 5.

    Supplementary Movie 5

    Movie of an untreated control well in the ETR assay.

  6. 6.

    Supplementary Movie 6

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

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DOI

https://doi.org/10.1038/nchembio.307

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