Breathing requires a complex activity pattern a neuronal network, located in medulla. In the ventral respiratory column the pre-Bötzinger complex (preBötC) appears to be most important. Glutamate modulates the activity of this network. While the role of astrocytes in glutamate clearance in the region has been investigated, studies on astroglial expression of metabotropic glutamate receptors in the pre-Bötzinger complex are missing so far. We used acute brainstem slices of mice that were loaded with Oregon Green to perform calcium imaging and 2-photon microscopy. Astrocytes were identified by the low K⁺ method (Dallwig et al., 2000) or by their green or red fluorescence in TgN(hGFAP-EGFP) and TgN(hGFAP-mRFP) mice, respectively. First, we tested the effect of group I mGluR agonist quisqualate. Quisqualate triggered calcium signals in almost every astrocyte in the VRG. Blockade of non-NMDA receptors with DNQX significantly reduced the amplitude of the calcium signal, indicating that also AMPA receptors are activated by quisqualate in astrocytes. We also demonstrated DHPG-mediated activation of group I mGluR in a majority of astrocytes. Astroglial calcium signals were not affected by TTX, but significantly reduced after depletion of intracellular calcium stores. Performing immunohistochemistry for mGluR1a, we proved that almost every astrocyte expressed mGluR1. While we demonstrated the functional expression of group I mGluR as well as non-NMDA receptors on astrocytes in the VRG, their physiological role remains to be clarified in future studies.