Detection of endogenous circadian rhythms of clock gene mRNA expression in mouse lung tissue using slice cultures

Detection of endogenous circadian rhythms in clock gene mRNA expression re-quires that mice be sacrificed at regular time intervals over one or more days. This protocol uses culture tissue slices obtained from a single mouse to collect time-course samples. We describe the procedure from preparation of lung slices to rhythmicity analysis of mRNA expression, including details to create hand-made culture inserts. This protocol is useful for many mammalian biological clock researchers because it allows a decrease in animal sacrifice. For complete details on the use and execution of this protocol, please refer to Matsumura et al. (2022).1