Journal of Neuroscience, Vol 5, 1471-1482, Copyright © 1985 by Society for Neuroscience
Development and subsequent neural tube effects on the excitability of cultured Xenopus myocytes
P DeCino and Y Kidokoro
We examined both the development of electrical excitability in cultured
Xenopus muscle over a period of 7 days, and the effects of neural tube on
the muscle action potential. During muscle development, delayed and
anomalous rectification were present in most cases within 24 hr. The action
potential was dependent on Na at all times examined, and the rate of rise
of the action potential (Vmax) increased substantially (seven-fold) from
the first 2 days to 6 to 7 days in vitro, reflecting an increase in Na
current density. In order to determine the mechanism for the increase in
Vmax, we examined single-channel Na currents using the gigaseal technique.
Single-channel conductance (gamma) did not increase substantially when
measured using the patch clamp technique: gamma = 24 pS at 1 to 2 days, and
gamma = 28 pS at 4 to 6 days. The channel open time at 14 degrees C was 0.6
msec for 1- to 2-day-old cells and 0.5 msec in 4- to 6-day-old cells at a
step potential 40 mV from rest. The time constant for current decay as well
as the time-to- peak current also did not change over time. Thus, channel
kinetics appear unchanged. The maximum inward current from summed records
was statistically greater for older cells, and the frequency of patches
displaying single-channel events increased from 75 to 98%. Thus, we
conclude that during development in vitro, Na current density increases as
a result of an increase in channel density without detectable alterations
in single-channel properties. Neural tube addition led to a further
increase in Vmax (two-fold), even in muscle cells with no apparent nerve
contact. Single channel analysis of cells in coculture revealed gamma to be
28 pS in three cells displaying a single amplitude peak for individual Na
currents. In the majority of cases (9/12), however, there appeared to be
two classes of Na channels present which were difficult to separate. The
larger conductance channel likely corresponds to the 28 pS class. The
smaller channels, when present, did not contribute substantially to the
population of events comprising the amplitude histogram. Other
single-channel kinetic parameters also did not change. We, therefore,
conclude that neural tube addition does not effect activation or
inactivation kinetics but likely causes a further increase in channel
density and possibly the induction of a second type of Na channel.