ELife assessment: Imaging Microglia Surveillance during Sleep-wake Cycles in Freely Behaving Mice

Abstract

Microglia surveillance manifests itself as dynamic changes in cell morphology and functional remodeling in response to fluctuations in the neural environment. Whether and how microglia surveillance is coupled to brain state switches during natural sleep-wake cycles, as well as under sleep deprivation, remain unclear. To address this question, we used miniature two-photon microscopy (mTPM) to acquire time-lapse high-resolution microglia images of the somatosensory cortex, along with EEG/EMG recordings and behavioral video, in freely-behaving mice. We uncovered fast and robust brain state-dependent changes in microglia surveillance, occurring in parallel with sleep dynamics and early-onset phagocytic microglial contraction during sleep deprivation stress. With the aid of the biosensor GRABNE2m, we also detected local norepinephrine fluctuation occurring in a sleep state-dependent manner. We showed that the locus coeruleus-norepinephrine system, which is crucial to sleep homeostasis, is required for both sleep state-dependent and stress-induced microglial responses and involves β2-adrenergic receptor signaling. These results provide direct evidence that microglial surveillance is exquisitely tuned to signals and stressors that regulate sleep dynamics and homeostasis so as to adjust its varied roles to complement those of neurons in the brain. In vivo imaging with mTPM in freely behaving animals, as demonstrated here, opens a new avenue for future investigation of microglia dynamics and sleep biology in freely behaving animals.