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Updated: Jul 5, 2026

In Vitro Bioluminescence Assay to Characterize Circadian Rhythm in Mammary Epithelial Cells
Published on: September 28, 2017
Circadian mPer1 gene expression in mesencephalic trigeminal nucleus cultures
Daniel J Hiler1, Aritra Bhattacherjee, Shin Yamazaki
1Department of Biological Sciences and the J.P. Scott Center for Neuroscience, Mind, and Behavior, Bowling Green State University, Bowling Green, OH 43403-0208, USA. dhiler@bgnet.bgsu.edu
Abstract:
The circadian timing system includes the major circadian pacemaker in the suprachiasmatic nucleus (SCN) of the hypothalamus and less well characterized circadian pacemakers in the brain and peripheral tissues throughout the body. The coupling between these discrete circadian clocks is not well understood, although individual neurons of the SCN are considered competent circadian pacemakers that interact to produce rhythms in the SCN and in its afferents. Because the SCN is a complex assemblage of small neurons of several phenotypes, we sought a simpler circadian brain nucleus with larger neurons that might provide insight into circadian timing not easily obtained from the SCN. Using bioluminescence imaging of brain tissue explants from transgenic mice containing the firefly luciferase gene luc controlled by the mPer1 promoter, we discovered elevated transgene expression throughout the mesencephalic trigeminal nucleus (Me5) of the brain stem. Large sensory neurons of the Me5 receive proprioceptive signals from periodontal ligaments and masseter muscle spindles. The Me5 cells displayed circadian rhythms with elevated expression in culture corresponding with the dark portion of the prior light cycle. Because of known interactions between the Me5 and the tuberomammillary nucleus and because of the role of both nuclei in satiety, it is possible that a circadian clock in the Me5 serves in regulating daily feeding behavior. This newly identified circadian pacemaker in the Me5 may prove useful for single-cell analyses of circadian gene expression in clock cells and for comparison with the SCN.
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