Department of Biochemistry and Molecular Pharmacology; Graduate School of Biomedical Sciences, Program in Neuroscience; Department of Neurobiology; Weaver Lab
Computational Neuroscience | Molecular and Cellular Neuroscience | Systems Biology
Circadian rhythms are driven by daily oscillations of gene expression. An important tool for studying cellular and tissue rhythms is the use of a gene reporter, such as bioluminescence from the reporter gene luciferase controlled by a rhythmically expressed gene of interest. Here we describe methods that allow measurement of bioluminescence from a freely-moving mouse housed in a standard cage. Using a LumiCycle In Vivo (Actimetrics), we determined conditions that allow detection of circadian rhythms of bioluminescence from the PER2 reporter, PER2::LUC, in freely behaving mice. We tested delivery of D-luciferin via a subcutaneous minipump and in the drinking water. Further, we demonstrate that a synthetic luciferase substrate, CycLuc1, can support circadian rhythms of bioluminescence, even when delivered at a lower concentration than D-luciferin. We share our analysis scripts and suggestions for further improvements in this method. This approach will be straightforward to apply to mice with tissue-specific reporters, allowing insights into responses of specific peripheral clocks to perturbations such as environmental or pharmacological manipulations.
bioluminescence, circadian, CycLuc1, in vivo, PERIOD2, peripheral oscillators, reporter gene, luciferase, systems biology, circadian rhythms, luciferase, neuroscience
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The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It is made available under a CC-BY-NC-ND 4.0 International license.
DOI of Published Version
bioRxiv 2020.08.24.264531; doi: https://doi.org/10.1101/2020.08.24.264531. Link to preprint on bioRxiv service.
Martin-Burgos B, Wang W, William I, Tir S, Mohammad I, Javed R, Smith College, Cui Y, Smith CB, van der Vinne V, Molyneux P, Miller SC, Weaver DR, Leise T, Harrington M. (2020). Methods for detecting PER2::LUCIFERASE bioluminescence rhythms in freely moving mice. University of Massachusetts Medical School Faculty Publications. https://doi.org/10.1101/2020.08.24.264531. Retrieved from https://escholarship.umassmed.edu/faculty_pubs/1769
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This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.