Volume 16, Number 16,
Issue of August 15, 1996
pp. 5117-5129
Copyright ©1996 Society for Neuroscience
Onset of Electrical Excitability during a Period of Circus Plasma
Membrane Movements in Differentiating Xenopus Neurons
Received April 5, 1996; revised June 3, 1996; accepted June 3, 1996.
Eric C. E. Olson
Department of Biology, University of California at San Diego, La
Jolla, California 92093
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 of
Xenopus 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.
Key words:
Xenopus spinal neurons;
circus movements;
lobopodia;
early neuronal marker;
sodium currents;
calcium currents;
potassium currents