CRAC channels regulate astrocyte Ca²⁺ signaling and gliotransmitter release to modulate hippocampal GABAergic transmission

Astrocytes are the major glial subtype in the brain and mediate numerous functions ranging from metabolic support to gliotransmitter release through signaling mechanisms controlled by Ca²⁺. Despite intense interest, the Ca²⁺ influx pathways in astrocytes remain obscure, hindering mechanistic insights into how Ca²⁺ signaling is coupled to downstream astrocyte-mediated effector functions. Here, we identified store-operated Ca²⁺ release-activated Ca²⁺ (CRAC) channels encoded by Orai1 and STIM1 as a major route of Ca²⁺ entry for driving sustained and oscillatory Ca²⁺ signals in astrocytes after stimulation of metabotropic purinergic and protease-activated receptors. Using synaptopHluorin as an optical reporter, we showed that the opening of astrocyte CRAC channels stimulated vesicular exocytosis to mediate the release of gliotransmitters, including ATP. Furthermore, slice electrophysiological recordings showed that activation of astrocytes by protease-activated receptors stimulated interneurons in the CA1 hippocampus to increase inhibitory postsynaptic currents on CA1 pyramidal cells. These results reveal a central role for CRAC channels as regulators of astrocyte Ca²⁺ signaling, gliotransmitter release, and astrocyte-mediated tonic inhibition of CA1 pyramidal neurons.