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.
Article: Live-cell imaging methods for the study of vagal afferents within the nucleus of the solitary tract.[show abstract] [hide abstract]
ABSTRACT: Substantial evidence suggests that vagal afferent functions are modulated by agonists acting on afferent terminals in the solitary nucleus (NST). Actions of these agonists are implied through intracellular recordings from cultured nodose ganglion cells or second order NST neurons. While high-quality data have been obtained using these methods, techniques in which physiological measurements can be made directly on the afferent terminal fields, in situ, in the NST, would eliminate several potential interpretive problems inherent in these less direct approaches. This paper describes methods developed to directly measure changes in presynaptic cytoplasmic calcium in vagal afferents using time-lapse laser confocal microscopy on the in vitro brainstem slice. Calcium green dextran (CG) transported from the nodose ganglion clearly demonstrates vagal afferent fibers ramifying throughout the NST in the in vitro brainstem slice. CG-labeled vagal afferents can be repeatedly activated by focal electrical stimulation, by agonists acting on presynaptic ligand-gated ion channels, and by molecules that are presumed to act directly on vagal afferents based on previous physiological and immunocytochemical studies. Image and preparation stability are a challenge to the success of the experiment; however, methods described here should assist direct studies of transduction events within other afferent terminal fields in the CNS.Journal of Neuroscience Methods 02/2006; 150(1):47-58. · 1.98 Impact Factor
Article: The astrocyte odyssey.[show abstract] [hide abstract]
ABSTRACT: Neurons have long held the spotlight as the central players of the nervous system, but we must remember that we have equal numbers of astrocytes and neurons in the brain. Are these cells only filling up the space and passively nurturing the neurons, or do they also contribute to information transfer and processing? After several years of intense research since the pioneer discovery of astrocytic calcium waves and glutamate release onto neurons in vitro, the neuronal-glial studies have answered many questions thanks to technological advances. However, the definitive in vivo role of astrocytes remains to be addressed. In addition, it is becoming clear that diverse populations of astrocytes coexist with different molecular identities and specialized functions adjusted to their microenvironment, but do they all belong to the umbrella family of astrocytes? One population of astrocytes takes on a new function by displaying both support cell and stem cell characteristics in the neurogenic niches. Here, we define characteristics that classify a cell as an astrocyte under physiological conditions. We will also discuss the well-established and emerging functions of astrocytes with an emphasis on their roles on neuronal activity and as neural stem cells in adult neurogenic zones.Progress in Neurobiology 11/2008; 86(4):342-67. · 8.87 Impact Factor
Article: Nicotinic cholinergic signaling in hippocampal astrocytes involves calcium-induced calcium release from intracellular stores[show abstract] [hide abstract]
ABSTRACT: In this report we provide evidence that neuronal nicotinic acetylcholine receptors (nAChRs) are present on hippocampal astrocytes and their activation produces rapid currents and calcium transients. Our data indicate that these responses obtained from astrocytes are primarily mediated by an AChR subtype that is functionally blocked by α-bungarotoxin (αBgt) and contains the α7 subunit (αBgt-AChRs). Furthermore, their action is unusual in that they effectively increase intracellular free calcium concentrations by activating calcium-induced calcium release from intracellular stores, triggered by influx through the receptor channels. These results reveal a mechanism by which αBgt-AChRs on astrocytes can efficiently modulate calcium signaling in the central nervous system in a manner distinct from that observed with these receptors on neurons.Proceedings of the National Academy of Sciences 04/2001; · 9.68 Impact Factor