Gap-Junctional Coupling between Neurons and Astrocytes in Primary Central Nervous System Cultures

Gap-junctional communication between neurons and astrocytes dissociated from rat brain was identified in culture by using dye-transfer assays and electrophysiological measurements. Cell types were identified by using antibodies against β -tubulin III, glial fibrillary acidic protein, and 2′,$3^{\pri...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1999-06, Vol.96 (13), p.7541-7546
Main Authors: Fróes, Maira M., Ana Helena P. Correia, Garcia-Abreu, José, Spray, David C., Antônio C. Campos de Carvalho, Neto, Vivaldo Moura
Format: Article
Language:English
Subjects:
Animals
Astrocytes
Astrocytes - cytology
Astrocytes - physiology
Biological Sciences
Biology
Brain
Cell Communication - physiology
Cells
Cells, Cultured
Central nervous system
Central Nervous System - cytology
Central Nervous System - physiology
Coculture Techniques
Cultured cells
Dyes
Electric current
Embryonic cells
Fluorescent Dyes
Gap Junctions - physiology
Isoquinolines
Nervous system
Neuroglia
Neurology
Neurons
Neurons - cytology
Neurons - physiology
Rats
Rats, Wistar
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Summary:Gap-junctional communication between neurons and astrocytes dissociated from rat brain was identified in culture by using dye-transfer assays and electrophysiological measurements. Cell types were identified by using antibodies against β -tubulin III, glial fibrillary acidic protein, and 2′,$3^{\prime}\text{-cyclic-nucleotide}$ phosphohydrolase, which are antigenic determinants of neurons, astroglia, and oligodendrocytes, respectively. Dye coupling was examined as a function of time after dissociated embryonic brain cells were plated onto confluent monolayers of postnatal astrocytes by intracellularly injecting the fluorochrome Lucifer yellow. Coupling of neurons to the astrocytic monolayer was most frequent between 48 h and 72 h in culture and declined over the next 4 days. This gradual uncoupling was accompanied by progressive neuronal maturation, as indicated by morphological measurements in camera lucida drawings. Dye spread was abolished reversibly by octanol, an agent that blocks gap junction channels in other systems. Double whole-cell voltage-clamp measurements confirmed the presence of heterocellular electrical coupling in these cocultures. Coupling was also seen between neurons and astrocytes in cocultures of cells dissociated from embryonic cerebral hemispheres but was rarely detectable in cocultures of postnatal brain cells. These data strongly suggest that junctional communication may provide metabolic and electrotonic interconnections between neuronal and astrocytic networks at early stages of neural development and that such interactions are weakened as differentiation progresses.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.96.13.7541