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Journal of Neuroscience, Vol 14, 6325-6335, Copyright © 1994 by Society for Neuroscience
Spontaneous neuronal calcium spikes and waves during early differentiation
X Gu, EC Olson and NC Spitzer
Department of Biology, University of California at San Diego, La Jolla 92093-0357.
Calcium ions play critical roles in neuronal development, but the factors
that govern spontaneous fluctuations in intracellular calcium are not well
understood. Transient, repeated elevations of calcium in embryonic Xenopus
spinal neurons have been recorded over periods of 1 hr in vitro and in
vivo, confocally imaging fluo-3-loaded cells at 5 sec intervals. Calcium
spikes and calcium waves are found both in neurons in culture and in the
intact spinal cord. Spikes rise rapidly to approximately 400% of baseline
fluorescence and have a characteristic double exponential decay, while
waves rise slowly to approximately 200% of baseline fluorescence and decay
slowly as well. Imaging of fura-2-loaded neurons indicates that
intracellular calcium increases from 50 to 500 nM during spikes. Both
spikes and waves are abolished by removal of extracellular calcium.
Developmentally, the incidence and frequency of spikes decrease while the
incidence and frequency of waves are constant. Spikes are generated by
spontaneous calcium-dependent action potentials that can be triggered by
low- threshold, T-type calcium current and are eliminated by agents that
block voltage-dependent calcium channels. They can be elicited by
depolarization, are generated in an all-or-none manner, and are rapidly and
bidirectionally propagated. Spikes also utilize intracellular calcium
stores, since blocking release from stores substantially reduces their
amplitude. Waves are not elicited by depolarization nor by activation of
glutamate receptors, and are propagated at a rate consistent with diffusion
of calcium. Waves are blocked by Ni2+ at a higher concentration than
required to block classical voltage-dependent calcium channels. Previous
work now suggests that spikes are required for expression of the
transmitter GABA and for potassium channel modulation. The present study
indicates that waves in growth cones are likely to regulate neurite
extension.
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