Real time, in vivo measurement of neuronal and peripheral clocks in Drosophila melanogaster
- Dalhousie University, Canada
- Westfälische Wilhelms University, Germany
- You.i Labs Inc, Canada
- 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
Article and author information
Author details
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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Real time, in vivo measurement of neuronal and peripheral clocks in Drosophila melanogaster
eLife 11:e77029.
https://doi.org/10.7554/eLife.77029
