Open AccessMethods for Detecting PER2:LUCIFERASE Bioluminescence Rhythms in Freely Moving Mice
Author(s) -
Blanca Martin-Burgos,
Wanqi Wang,
Ivana William,
Selma Tir,
Innus Mohammad,
Reja Javed,
S Kendall Smith,
Yilin Cui,
Jessica Arzavala,
Dalilah Mora,
Corey Smith,
Vincent van der Vinne,
Penny C. Molyneux,
Stephen J. Miller,
David R. Weaver,
Tanya Leise,
Mary E. Harrington
Publication year - 2021
Publication title -
journal of biological rhythms
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.484
H-Index - 101
eISSN - 1552-4531
pISSN - 0748-7304
DOI - 10.1177/07487304211062829
Subject(s) - per2 , bioluminescence , luciferase , circadian rhythm , luciferin , bioluminescence imaging , reporter gene , biology , rhythm , clock , circadian clock , microbiology and biotechnology , neuroscience , gene expression , gene , medicine , transfection , biochemistry
Circadian rhythms are driven by daily oscillations of gene expression. An important tool for studying cellular and tissue circadian 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 circadian 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. The LumiCycle In Vivo applies a background subtraction that corrects for effects of room temperature on photomultiplier tube (PMT) output. We tested delivery of d-luciferin via a subcutaneous minipump and in the drinking water. We demonstrate spikes in bioluminescence associated with drinking bouts. 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, and can support longer-term studies. A small difference in phase of the PER2::LUC bioluminescence rhythms, with females phase leading males, can be detected with this technique. 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.