Norepinephrine-induced calcium signaling in astrocytes in the respiratory network of the ventrolateral medulla

• Norepinephrine modulates calcium signaling in astrocytes of the respiratory network.
• Norepinephrine acts via the G protein coupled Phospholipase C-Inositol trisphosphate cascade (Gq-PLC-IP3).
• Astrocytes that express the genetically encoded calcium sensor GCaMP3 controlled by the Glast locus allow unequivocal identification of astrocytic calcium signals.
• Norepinephrine-induced signaling in astrocytes has to be considered a key modulator of neuronal activity in the respiratory network.

The neuronal activity in the respiratory network of the ventrolateral medulla strongly depends on a variety of different neuromodulators. Since the respiratory activity generated by neurons in the pre-Bötzinger complex (preBötC) is stabilized by astrocytes, we investigated potential effects of the neuromodulator norepinephrine (NE) on the astrocytic calcium signaling in the ventral respiratory group. In acutely isolated brainstem slices from wild type mice (postnatal day 1–10) we performed calcium imaging experiments using Oregon Green 488 BAPTA-1 AM as a calcium indicator dye. Astrocytes in the preBötC, which were identified by their unique intracellular calcium rise after the reduction of the extracellular K+ concentration, showed calcium rises in response to norepinephrine. These calcium signals persisted after blockade of neuronal activity by tetrodotoxin (TTX) indicating that they were independent of neuronal activity. Furthermore, application of the endoplasmic reticulum calcium pump blocker cyclopiazonic acid (CPA) diminished norepinephrine-induced calcium signals. This results could be confirmed using transgenic mice with astrocyte specific expression of GCaMP3. Thus, norepinephrine might, apart from acting directly on neurons, influence and modulate respiratory network activity via the modulation of astroglial calcium signaling.