Small-molecule CEM3 strengthens single-cell oscillators in the suprachiasmatic nucleus

A robust endogenous clock is required for proper function of many physiological processes. The suprachiasmatic nucleus (SCN) constitutes our central circadian clock and allows us to adapt to daily changes in the environment. Aging can cause a decline in the amplitude of circadian rhythms in SCN and peripheral clocks, which contributes to increased risk of several chronic diseases. Strengthening clock function would therefore be an effective strategy to improve health. A high-throughput chemical screening has identified clock-enhancing molecule 3 (CEM3) as small molecule that increases circadian rhythm amplitude in cell lines and SCN explants. It is, however, currently not known whether CEM3 acts by enhancing the amplitude of individual single-cell oscillators or by enhancing synchrony among neurons. In view of CEM3's potential, it is of evident importance to clarify the mode of action of CEM3. Here, we investigated the effects of CEM3 on single-cell PERIOD2::LUCIFERASE rhythms in mouse SCN explants. CEM3 increased the amplitude in approximately 80%-90% of the individual cells in the SCN without disrupting the phase and/or period of their rhythms. Noticeably, CEM3's effect on amplitude is independent of the cell's initial amplitude. These findings make CEM3 a potential therapeutic candidate to restore compromised amplitude in circadian rhythms and will boost the development of other molecular approaches to improve health. Circadian rhythms are generated by the suprachiasmatic nucleus (SCN). Disruption of rhythms is a causal risk factor for the development of diseases, like depression or immune disorders. The small-molecule CEM3 strengthens SCN rhythms by enhancement of amplitude. We asked whether amplitude enhancement is a single-cell or a network phenomenon. The results indicate that CEM3 operates by increasing the amplitude of individual SCN neurons. This makes CEM3 an interesting therapeutic candidate for improving health by strengthening our internal clock.image