Effects of patterned electrical activity on neurite outgrowth from mouse sensory neurons

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.