Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

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

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.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

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.

References

  1. 1

    Ayd, F.J. & Blackwell, B. Discoveries in Biological Psychiatry (Lippincott, Philadelphia, 1970).

  2. 2

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

    CAS  Article  Google Scholar 

  3. 3

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

    CAS  Article  Google Scholar 

  4. 4

    Lehár, J., Stockwell, B.R., Giaever, G. & Nislow, C. Combination chemical genetics. Nat. Chem. Biol. 4, 674–681 (2008).

    Article  Google Scholar 

  5. 5

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

    CAS  Article  Google Scholar 

  6. 6

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

    CAS  Article  Google Scholar 

  7. 7

    Hert, J., Keiser, M., Irwin, J., Oprea, T. & Shoichet, B. Quantifying the relationships among drug classes. J. Chem. Inf. Model. 48, 755–765 (2008).

    CAS  Article  Google Scholar 

  8. 8

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

    CAS  Article  Google Scholar 

  9. 9

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

    CAS  Article  Google Scholar 

  10. 10

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

    CAS  Article  Google Scholar 

  11. 11

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

    CAS  Article  Google Scholar 

  12. 12

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

    CAS  Article  Google Scholar 

  13. 13

    Thase, M.E., Trivedi, M.H. & Rush, A.J. MAOIs in the contemporary treatment of depression. Neuropsychopharmacology 12, 185–219 (1995).

    CAS  Article  Google Scholar 

  14. 14

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

    Article  Google Scholar 

  15. 15

    Meng, X., Noyes, M.B., Zhu, L.J., Lawson, N.D. & Wolfe, S.A. Targeted gene inactivation in zebrafish using engineered zinc-finger nucleases. Nat. Biotechnol. 26, 695–701 (2008).

    CAS  Article  Google Scholar 

  16. 16

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

    CAS  Article  Google Scholar 

  17. 17

    Rovida, C. & Hartung, T. 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).

    Article  Google Scholar 

  18. 18

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

    CAS  Article  Google Scholar 

  19. 19

    James, C.A., Weininger, D. & Delany, J. Daylight theory manual (Daylight Chemical Information Systems, 1995).

  20. 20

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

    Google Scholar 

Download references

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

Authors

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.

Ethics declarations

Competing interests

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)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

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

Download citation

Further reading

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing