Real time, in vivo measurement of neuronal and peripheral clocks in Drosophila melanogaster

  1. Peter S Johnstone
  2. Maite Ogueta
  3. Olga Akay
  4. Inan Top
  5. Sheyum Syed
  6. Ralf Stanewsky
  7. Deniz Top  Is a corresponding author
  1. Dalhousie University, Canada
  2. Westfälische Wilhelms University, Germany
  3. You.i Labs Inc, Canada
  4. University of Miami, United States

Abstract

Circadian clocks are highly conserved transcriptional regulators that control ~24-hour oscillations in gene expression, physiological function, and behavior. Circadian clocks exist in almost every tissue and are thought to control tissue-specific gene expression and function, synchronized by the brain clock. Many disease states are associated with loss of circadian regulation. How and when circadian clocks fail during pathogenesis remains largely unknown because it is currently difficult to monitor tissue-specific clock function in intact organisms. Here, we developed a method to directly measure the transcriptional oscillation of distinct neuronal and peripheral clocks in live, intact Drosophila, which we term Locally Activatable BioLuminescence, or LABL. Using this method, we observed that specific neuronal and peripheral clocks exhibit distinct transcriptional properties. Loss of the receptor for PDF, a circadian neurotransmitter critical for the function of the brain clock, disrupts circadian locomotor activity but not all tissue-specific circadian clocks. We found that, while peripheral clocks in non-neuronal tissues were less stable after the loss of PDF signaling, they continued to oscillate. We also demonstrate that distinct clocks exhibit differences in their loss of oscillatory amplitude or their change in period, depending on their anatomical location, mutation, or fly age. Our results demonstrate that LABL is an effective tool that allows rapid, affordable, and direct real-time monitoring of individual clocks in vivo.

Data availability

The codes used in data analysis can be found at https://github.com/deniztop/LABLAll data points used in generating the figures can be found at Dryad

The following data sets were generated

Article and author information

Author details

  1. Peter S Johnstone

    Department of Pediatrics, Dalhousie University, Halifax, Canada
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9421-811X
  2. Maite Ogueta

    Institute of Neuro- and Behavioral Biology, Westfälische Wilhelms University, Münster, Germany
    Competing interests
    The authors declare that no competing interests exist.
  3. Olga Akay

    Department of Pediatrics, Dalhousie University, Halifax, Canada
    Competing interests
    The authors declare that no competing interests exist.
  4. Inan Top

    You.i Labs Inc, Ottawa, Canada
    Competing interests
    The authors declare that no competing interests exist.
  5. Sheyum Syed

    Department of Physics, University of Miami, Miami, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Ralf Stanewsky

    Institute of Neuro- and Behavioral Biology, Westfälische Wilhelms University, Münster, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8238-6864
  7. Deniz Top

    Department of Pediatrics, Dalhousie University, Halifax, Canada
    For correspondence
    dtop@dal.ca
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1042-8460

Funding

Natural Sciences and Engineering Research Council of Canada (RGPIN-2019-06101)

  • Deniz Top

National Science Foundation (IOS 1656603)

  • Sheyum Syed

Deutsche Forschungsgemeinschaft (INST 211/835-1 FUGG)

  • Ralf Stanewsky

Deutsche Forschungsgemeinschaft (STA421/7-1)

  • Ralf Stanewsky

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

© 2022, Johnstone et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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https://doi.org/10.7554/eLife.77029

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