Methods for Detecting PER2:LUCIFERASE Bioluminescence Rhythms in Freely Moving Mice

Authors

Blanca Martin-Burgos, Smith College
Wanqi Wang, Smith College
Ivana William, Smith College
Selma Tir, Smith College
Innus Mohammad, University of Massachusetts Chan Medical School
Reja Javed, Smith College
Stormi Smith, Smith College
Yilin Cui, Smith College
Jessica Arzavala, Smith College
Dalilah Mora, Smith College
Ciearra B. Smith, University of Massachusetts Chan Medical School
Vincent van der Vinne, University of Massachusetts Chan Medical School
Penny C. Molyneux, Smith College
Stephen C. Miller, University of Massachusetts Chan Medical School
David R. Weaver, University of Massachusetts Chan Medical School
Tanya L. Leise, Amherst College
Mary E. Harrington, Smith CollegeFollow

Document Type

Article

Publication Date

2-1-2022

Publication Title

Journal of Biological Rhythms

Abstract

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.

Keywords

bioluminescence, circadian, CycLuc1, in vivo, luciferase, PERIOD2, peripheral oscillators, reporter gene

Volume

37

Issue

1

First Page

78

Last Page

93

DOI

10.1177/07487304211062829

ISSN

07487304

Rights

© 2021 The Author(s)

Comments

Archived as published. Open access article.

Recommended Citation

Martin-Burgos, Blanca; Wang, Wanqi; William, Ivana; Tir, Selma; Mohammad, Innus; Javed, Reja; Smith, Stormi; Cui, Yilin; Arzavala, Jessica; Mora, Dalilah; Smith, Ciearra B.; van der Vinne, Vincent; Molyneux, Penny C.; Miller, Stephen C.; Weaver, David R.; Leise, Tanya L.; and Harrington, Mary E., "Methods for Detecting PER2:LUCIFERASE Bioluminescence Rhythms in Freely Moving Mice" (2022). Psychology: Faculty Publications, Smith College, Northampton, MA.
https://scholarworks.smith.edu/psy_facpubs/87