Postsynaptic competition between calcineurin and PKA regulates mammalian sleep–wake cycles

• Published in: Nature (London) 2024-12, Vol.636 (8042), p.412-421
• Main Authors: Wang, Yimeng, Cao, Siyu, Tone, Daisuke, Fujishima, Hiroshi, Yamada, Rikuhiro G., Ohno, Rei-ichiro, Shi, Shoi, Matsuzawa, Kyoko, Yada, Saori, Kaneko, Mari, Sakamoto, Hirokazu, Onishi, Taichi, Ukai-Tadenuma, Maki, Ukai, Hideki, Hanashima, Carina, Hirose, Kenzo, Kiyonari, Hiroshi, Sumiyama, Kenta, Ode, Koji L., Ueda, Hiroki R.
• Format: Article
• Language: English
• Subjects: 45/41; 631/337/458/1733; 631/378/1385/519; 64/60; Animals; Calcineurin - deficiency; Calcineurin - genetics; Calcineurin - metabolism; CRISPR-Cas Systems - genetics; Cyclic AMP-Dependent Protein Kinases - chemistry; Cyclic AMP-Dependent Protein Kinases - deficiency; Cyclic AMP-Dependent Protein Kinases - genetics; Cyclic AMP-Dependent Protein Kinases - metabolism; Dependovirus - genetics; Dependovirus - metabolism; Female; Humanities and Social Sciences; Humans; Male; Mice; Mice, Knockout; multidisciplinary; Phosphorylation; Protein Phosphatase 1 - chemistry; Protein Phosphatase 1 - deficiency; Protein Phosphatase 1 - genetics; Protein Phosphatase 1 - metabolism; Science; Science (multidisciplinary); Sleep - genetics; Sleep - physiology; Sleep Duration; Synapses - metabolism; Time Factors; Wakefulness - physiology
• ISSN: 0028-0836; 1476-4687; 1476-4687
• DOI: 10.1038/s41586-024-08132-2

The phosphorylation of synaptic proteins is a significant biochemical reaction that controls the sleep–wake cycle in mammals 1 , 2 – 3 . Protein phosphorylation in vivo is reversibly regulated by kinases and phosphatases. In this study, we investigate a pair of kinases and phosphatases that reciprocally regulate sleep duration. First, we perform a comprehensive screen of protein kinase A (PKA) and phosphoprotein phosphatase (PPP) family genes by generating 40 gene knockout mouse lines using prenatal and postnatal CRISPR targeting. We identify a regulatory subunit of PKA ( Prkar2b ), a regulatory subunit of protein phosphatase 1 (PP1; Pppr1r9b ) and catalytic and regulatory subunits of calcineurin (also known as PP2B) ( Ppp3ca and Ppp3r1 ) as sleep control genes. Using adeno-associated virus (AAV)-mediated stimulation of PKA and PP1–calcineurin activities, we show that PKA is a wake-promoting kinase, whereas PP1 and calcineurin function as sleep-promoting phosphatases. The importance of these phosphatases in sleep regulation is supported by the marked changes in sleep duration associated with their increased and decreased activities, ranging from approximately 17.3 h per day (PP1 expression) to 4.3 h per day (postnatal CRISPR targeting of calcineurin). Localization signals to the excitatory post-synapse are necessary for these phosphatases to exert their sleep-promoting effects. Furthermore, the wake-promoting effect of PKA localized to the excitatory post-synapse negated the sleep-promoting effect of PP1–calcineurin. These findings indicate that PKA and PP1–calcineurin have competing functions in sleep regulation at excitatory post-synapses. Calcineurin–PP1 and PKA acting at excitatory post-synapses reciprocally regulate sleep duration in mice, whereby PKA promotes wakefulness and calcineurin–PP1 promote sleep.