Methods for detecting PER2::LUCIFERASE bioluminescence rhythms in freely moving mice [preprint]
Martin-Burgos, B. ; Wang, W. ; William, I. ; Tir, S. ; Mohammad, I. ; Javed, R. ; Smith College ; Cui, Y. ; Smith, Ciearra B. ; van der Vinne, V. ... show 5 more
Citations
Student Authors
Faculty Advisor
Academic Program
Document Type
Publication Date
Subject Area
Embargo Expiration Date
Link to Full Text
Abstract
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.
Source
bioRxiv 2020.08.24.264531; doi: https://doi.org/10.1101/2020.08.24.264531. Link to preprint on bioRxiv service.
Year of Medical School at Time of Visit
Sponsors
Dates of Travel
DOI
Permanent Link to this Item
PubMed ID
Other Identifiers
Notes
Funding and Acknowledgements
Corresponding Author
Related Resources
Now published in Journal of Biological Rhythms, doi: https://doi.org/10.1177/07487304211062829.