Rapid communication between neurons and astrocytes in primary cortical cultures.
The identification of neurotransmitter receptors and voltage-sensitive ion channels on astrocytes (reviewed by Barres, 1991) has renewed interest in how these cells respond to neuronal activity. To investigate the physiology of neuron astrocyte signaling, we have employed primary cortical cultures that contain both neuronal and glial cells. As the neurons in these cultures exhibit synchronous spontaneous synaptic activity, we have used both calcium imaging and whole-cell recording techniques to identify physiological activity in astrocytes related to neuronal activity. Whole-cell voltage-clamp records from astrocytes revealed rapid inward currents that coincide with bursts of electrical activity in neighboring neurons. Calcium imaging studies demonstrate that these currents in astrocytes are not always associated with slowly propagating calcium waves. Inclusion of the dye Lucifer yellow within patch pipettes confirmed that astrocytes are extensively coupled to each other but not to adjacent neurons, indicating that the currents observed are not due to gap junction connections between these cell types. These currents do not reflect widespread diffusion of glutamate or potassium released during neuronal activity since a population of small, round, multipolar presumed glial cells that are not dye coupled to adjacent cells did not display electrical currents coincident with neuronal firing, even though they respond to locally applied glutamate and potassium. These findings indicate that, in addition to the relatively slow signaling conveyed by calcium waves, astrocytes also display rapid electrical responses to neuronal activity.
Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
T H
TH
Murphy
L A
LA
Blatter
W G
WG
Wier
J M
JM
Baraban
D000818:Animals; D001253:Astrocytes / Q000378:metabolism / Q000502:physiology*; D002118:Calcium / Q000037:antagonists & inhibitors / Q000502:physiology; D002450:Cell Communication; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology*; D004558:Electric Stimulation; D004594:Electrophysiology; D005456:Fluorescent Dyes; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D007365:Intercellular Junctions / Q000502:physiology; D007546:Isoquinolines; D009474:Neurons / Q000502:physiology*; D013779:Tetrodotoxin / Q000494:pharmacology
D005456:Fluorescent Dyes; D005904:Glial Fibrillary Acidic Protein; D007546:Isoquinolines; D013779:Tetrodotoxin; C017475:lucifer yellow; D002118:Calcium