Gi/o protein‐coupled receptors inhibit neurons but activate astrocytes and stimulate gliotransmission

G protein‐coupled receptors (GPCRs) play key roles in intercellular signaling in the brain. Their effects on cellular function have been largely studied in neurons, but their functional consequences on astrocytes are less known. Using both endogenous and chemogenetic approaches with DREADDs, we have investigated the effects of Gq and Gi/o GPCR activation on astroglial Ca2+‐based activity, gliotransmitter release, and the functional consequences on neuronal electrical activity. We found that while GqGPCR activation led to cellular activation in both neurons and astrocytes, Gi/oGPCR activation led to cellular inhibition in neurons and cellular activation in astrocytes. Astroglial activation by either Gq or Gi/o protein‐mediated signaling stimulated gliotransmitter release, which increased neuronal excitability. Additionally, activation of Gq and Gi/o DREADDs in vivo increased astrocyte Ca2+ activity and modified neuronal network electrical activity. Present results reveal additional complexity of the signaling consequences of excitatory and inhibitory neurotransmitters in astroglia‐neuron network operation and brain function. Main Points Gq signaling in neurons and astrocytes leads to cellular activation, but while Gi/o signaling in neurons leads to cellular inhibition, it leads to activation in astrocytes and downstream gliotransmission that modulates neuronal activity.