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Journal of Neuroscience, Vol 10, 2950-2964, Copyright © 1990 by Society for Neuroscience
Effects of patterned electrical activity on neurite outgrowth from mouse sensory neurons
RD Fields, EA Neale and PG Nelson
Laboratory of Developmental Neurobiology, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892.
A noninvasive method of electric stimulation was used in cell culture
preparations to determine the effects of patterned electrical activity on
the morphology and motility of mammalian central nervous system growth
cones. Neurites from dorsal root ganglion (DRG) neurons of fetal mice were
allowed to grow under the barrier of an insert placed in culture dishes.
The insert confined the cell bodies within separate experimental and
control compartments, and provided a means of exciting action potentials in
the growing neurites by extracellular current pulses delivered across the
barrier. A phasic pattern of stimulation caused immediate retraction of the
filopodia and lamellipodium. Further outgrowth was halted and in many cases
retraction of the neurite ensued. No changes in morphology or growth cone
motility were evoked by electric stimulation when action potentials were
blocked with 1 microM tetrodotoxin (TTX). These effects depended on the
rate, pattern, and duration of stimulation. Phasic stimulation was more
effective than stimulation with the same number of impulses delivered at a
constant frequency. An important new observation was that cultures exposed
to phasic stimulation for several hours contained actively growing neurites
with normal growth cones which were insensitive to the stimulus. This
apparent accommodation in neurites exposed to chronic stimulation may
involve processes that regulate calcium conductance or buffering. Cessation
of neurite outgrowth by action potentials could represent one mechanism
linking morphological and functional characteristics in the developing CNS
of mammals, by stabilizing the outgrowth of neurites forming appropriate
synaptic contacts and leading to the retraction of growth cones from
collaterals that have not formed appropriate contacts at the time the
neuron enters into a functionally active circuit.
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