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Living neurons are usually first identifiable in primary cultures at the time of neurite initiation, and studies of excitability have been restricted largely to the subsequent period. A morphological early marker is described that identifies neurons for whole-cell voltage-clamp recordings before neurite initiation. Video time-lapse recordings of cultured cells dissociated from neurectoderm ofXenopus neural plate stage embryos reveal cells demonstrating circus movements, in which blebs of plasma membrane propagate around the cell circumference within a period of several minutes. All neurons demonstrate circus movements before morphological differentiation; the fraction of cells exhibiting circus movements that differentiate morphologically depends on the substrate on which they are cultured. Blockade of circus activity with cytochalasin B does not prevent neuronal differentiation. Circus movements are not neurectoderm-specific because they similarly predict differentiation of myocytes developing in mesodermal cultures. Initially inexcitable, neurons develop voltage-dependent K+, Na+, and Ca2+ currents during the period of several hours in which they exhibit circus movements. The early development of depolarization-induced elevations of [Ca2+]i several hours before morphological differentiation corresponds to the previously described onset of functionally significant spontaneous elevations of [Ca2+]i in these neurons and demonstrates a role for early expression of voltage-dependent ion channels.

Onset of Electrical Excitability during a Period of Circus Plasma Membrane Movements in DifferentiatingXenopusNeurons