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Journal of Neuroscience, Vol 12, 4834-4845, Copyright © 1992 by Society for Neuroscience
Spontaneous synchronous synaptic calcium transients in cultured cortical neurons
TH Murphy, LA Blatter, WG Wier and JM Baraban
Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
The firing pattern displayed by neuronal aggregates is thought to play a
key role in cortical development and physiology. In this study, we have
employed optical recording of intracellular calcium to monitor activity of
multiple neurons simultaneously in primary cortical cultures. With this
approach, we have observed spontaneous synchronous calcium transients among
adjacent cortical neurons. These transients appear to be mediated by
prominent spontaneous synaptic excitation, as they are enhanced by
picrotoxin, a blocker of inhibitory GABAergic transmission, and reduced by
antagonism of glutamate receptors or addition of TTX. After picrotoxin
treatment, the calcium transients exhibit regular frequency and amplitude,
and occur in synchrony with bursts of excitatory synaptic potentials every
10-20 sec. Using electrical stimulation, we have identified a relative
refractory period, extending up to 5 sec after a synchronous burst, that
may play a role in cell synchronization. NMDA receptor antagonists or
reduced extracellular calcium levels lower the amplitude of the calcium
transients yet fail to alter their frequency, suggesting that intracellular
calcium levels may not be a major determinant of burst frequency. In
contrast, mild depolarization with kainic acid (0.5-1 microM) increased
burst frequency up to fivefold, suggesting a critical dependence of
rhythmic activity on membrane potential. Chronic blockade of electrical
activity with TTX beginning a few days after plating of cultures dampens
the amplitude and significantly increases the frequency of calcium
transients in mature cultures. These studies demonstrate that aggregates of
cultured cortical neurons express synchronous firing activity in vitro and
that this network activity is dependent in part on neuronal firing during
development.
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