Microglia Regulate Sleep via Calcium-Dependent Modulation of Norepinephrine Transmission

Sleep interacts reciprocally with immune system activity, but its specific relationship with microglia – the resident immune cells in the brain – remains poorly understood. Here we show that microglia can regulate sleep through a mechanism involving Gi-coupled GPCRs, intracellular Ca²⁺signaling, and suppression of norepinephrine transmission. Chemogenetic activation of microglia Gi signaling strongly promoted sleep, whereas pharmacological blockade of Gi-coupled P2Y12 receptors decreased sleep. Two-photon imaging showed that P2Y12/Gi activation elevated microglia intracellular Ca²⁺, and blockade of this Ca²⁺elevation largely abolished the Gi-induced sleep increase. Microglia Ca²⁺level also increased at natural wake-to-sleep transitions, caused partly by reduced norepinephrine. Furthermore, imaging of norepinephrine activity with its biosensor showed that microglia P2Y12/Gi activation significantly reduced norepinephrine, partly by increasing the adenosine concentration. Thus, microglia can regulate sleep through reciprocal interactions with norepinephrine transmission.