Methods Mol Biol. 2022;2482:217-242. doi: 10.1007/978-1-0716-2249-0_15.
There is high interest in investigating the daily dynamics of gene expression in mammalian organs, for example, in liver. Such studies help to elucidate how and with what kinetics peripheral clocks integrate circadian signals from the suprachiasmatic nucleus, which harbors the circadian master pacemaker, with other systemic and environmental cues, such as those associated with feeding and hormones. Organ sampling around the clock, followed by the analysis of RNA and/or proteins, is the most commonly used procedure in assessing rhythmic gene expression. However, this method requires large cohorts of animals and is only applicable to behaviorally rhythmic animals whose phases are known. Real-time recording of gene expression rhythms using luciferase reporters has emerged as a powerful method to acquire continuous, high-resolution datasets from freely moving individual mice. Here, we share our experience and protocols with this technique, using the RT-Biolumicorder setup.
Keywords: Circadian rhythms; Feeding rhythms; Gene expression regulation; Luciferase; Mouse liver; Peripheral oscillators; RT-Biolumicorder; Real-time bioluminescence recording; Transcription.
- PMID: 35610430
- DOI: 10.1007/978-1-0716-2249-0_15