Why do we sleep? — The Role of Calcium and Phosphorylation in Sleep

Sleep remains one of greatest remaining mysteries. At the Sleep 2012 conference, we conceived a shift from the concept of “sleep substances” to “wake substances” such as calcium, suggesting that sleep homeostasis may arise from the integration of wake-related activity. Inspired by Dr. Setsuro Ebashi’s work on calcium signaling, we investigated calcium’s role in sleep regulation.

Using our Triple-CRISPR method (Sunagawa et al. 2016), we screened 25 genes related to calcium channels and pumps, revealing calcium as a brake on brain activity to promote sleep (Tatsuki et al. 2016). We also developed a tissue-clearing method CUBIC (Susaki et al. 2014; Tainaka et al. 2014) to visualize calcium’s effects on neural circuits. Further work showed that calcium-dependent enzymes, CaMKIIα/β kinases, act as calcium “memory” devices, with phosphorylation sites controlling sleep onset, duration, and termination (Tone et al. 2022). Other direct and indirect calcium-dependent phosphatases, Calcineurin and PP1 (sleep-promoting), and opposing kinases, PKA (wake-promoting), function as synaptic sleep switches (Wang et al. 2024).

We also identified the ryanodine receptor 1, a calcium channel, as a molecular target of inhalational anesthetics, hinting at shared pathways between anesthesia and sleep (Kanaya et al. 2025). Lastly, we proposed the WISE (Wake Inhibition Sleep Enhancement) mechanism, where quiet wakefulness suppresses and deep sleep strengthens synaptic connections, explaining links between sleep, depression, and antidepressant effects (Kinoshita et al. 2025).