Vagal afferent stimulation activates astrocytes in the nucleus of the solitary tract via AMPA receptors: evidence of an atypical neural-glial interaction in the brainstem.

Abstract

The nucleus of the solitary tract (NST), located in the dorsomedial medulla, is the site of visceral sensory modulation of a variety of homeostatic reflexes. Given recent advancements in the understanding of active regulation of synaptic information flow by astrocytes, we sought to determine whether afferent sensory inputs to NST neurons also activates NST astrocytes. Using confocal, live-cell calcium imaging of brainstem slices, we investigated the possibility that stimulation of vagal sensory afferents, the major sensory input into the NST, activated NST astrocytes, as indicated by increases in astrocytic intracellular calcium concentrations ([Ca²⁺](i)). Astrocytes and neurons were preloaded with the calcium reporter dye Calcium Green, and astrocytes were selectively stained by sulforhodamine 101. Electrical stimulation of vagal afferent axons produced rapid increases in [Ca²⁺](i) in NST astrocytes as well as neurons. Surprisingly, this effect on astrocytes was blocked by the AMPA receptor antagonist NBQX and was unaffected by antagonism of NMDA and metabotropic glutamate receptors. Bath application of AMPA also activated astrocytes. This activation was dependent on extracellular Ca²⁺ influx through both typical AMPA receptors and calcium-permeable AMPA receptors. This AMPA-mediated Ca²⁺ influx was further amplified by actions of the ryanodine receptor by way of calcium-induced calcium release. Our immunohistochemical staining of NST cells further verified the presence of the AMPAR subunit GluR1 on astrocytes. These observations suggest that NST astrocytes may be active participants in the regulation of autonomic reflexes even in the normal, healthy state.