Identification of pathways that regulate circadian rhythms using a larval zebrafish small molecule screen
Abstract
The circadian clock ensures that behavioral and physiological processes occur at appropriate times during the 24-hour day/night cycle, and is regulated at both the cellular and organismal levels. To identify pathways acting on intact animals, we performed a small molecule screen using a luminescent reporter of molecular circadian rhythms in zebrafish larvae. We identified both known and novel pathways that affect circadian period, amplitude and phase. Several drugs identified in the screen did not affect circadian rhythms in cultured cells derived from luminescent reporter embryos or in established zebrafish and mammalian cell lines, suggesting they act via mechanisms absent in cell culture. Strikingly, using drugs that promote or inhibit inflammation, as well as a mutant that lacks microglia, we found that inflammatory state affects circadian amplitude. These results demonstrate a benefit of performing drug screens using intact animals and provide novel targets for treating circadian rhythm disorders.
Additional Information
© 2019 The Author(s). This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Received 21 March 2018; Accepted 31 July 2019; Published 27 August 2019. Data Availability: The datasets generated and analyzed in the current study are available from the corresponding author on reasonable request. We thank Daisy Chilin, Viveca Sapin, Ayato Sato and Natsuko Ono for technical assistance, and William Hurst and Sanofi-Aventis for providing reagents. This work was supported by grants from the NIH (NS070911, NS101158, NS095824, NS101665) and Sanofi-Aventis to D.A.P., and the JSPS (15H05590) to T.H. Author Contributions: E.A.M. and D.A.P. designed research. E.A.M., S.L., M.H. and A.W. performed the screen. E.A.M. and C.S. performed additional experiments. E.A.M. and C.N.C. analyzed data. T.K.T. and T.H. performed cell culture experiments. A.G. provided small molecules. E.A.M. and D.A.P. wrote the paper, with input from T.K.T., T.H. and S.A.K. The authors declare no competing interests.Attached Files
Published - s41598-019-48914-7.pdf
Supplemental Material - 41598_2019_48914_MOESM1_ESM.pdf
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Additional details
- PMCID
- PMC6712016
- Eprint ID
- 98268
- Resolver ID
- CaltechAUTHORS:20190827-104428064
- NS070911
- NIH
- NS101158
- NIH
- NS095824
- NIH
- NS101665
- NIH
- Sanofi-Aventis
- 15H05590
- Japan Society for the Promotion of Science (JSPS)
- Created
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2019-08-27Created from EPrint's datestamp field
- Updated
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2021-11-16Created from EPrint's last_modified field