Journal of Neuroscience, Vol 4, 1453-1467, Copyright © 1984 by Society for Neuroscience
Identification and characterization of voltage-sensitive calcium channels in neuronal clonal cell lines
SB Freedman, G Dawson, ML Villereal and RJ Miller
Voltage-sensitive calcium channels ( VSCCs ) have been identified in three
clonal cells. These are the neuroblastoma X Chinese hamster brain hybrid (
NCB -20), the neuroblastoma X glioma hybrid (NG108-15), and the
neuroblastoma ( N4TG1 ). Depolarization of NCB -20 cells with 50 mM KCl or
50 microM veratridine (VE) produced a 2- to 3-fold increase in net 45Ca2+
uptake. In NCB -20 cells, this voltage-sensitive 45Ca2+ uptake was
inhibited selectively by organic calcium antagonists such as nitrendipine,
cinnarizine, verapamil, and diltiazem (IC50 values = 6.4, 750, 1800, and
4500 nM, respectively). High K+-induced uptake was unaffected by
4-aminopyridine, tetraethylammonium, and tetrodotoxin (TTX), whereas
VE-induced 45Ca2+ uptake was completely blocked by 3 microM TTX. In
contrast to NCB -20 cells, NG108-15 cells showed a much smaller response to
depolarizing stimuli. Following differentiation of NG108-15 cells by
chronic treatment with 10 microM prostaglandin E1 and 50 microM
3-isobutyl-1-methylxanthine, depolarization induced a large increase in
voltage-sensitive 45Ca2+ uptake. This induction was apparent after 24 hr
and increased linearly for 96 hr. VSCC activity was also induced by 1.5%
dimethyl sulfoxide and by other agents that increase intracellular cAMP,
such as forskolin (1 microM) and cholera toxin (1 microgram/ml).
Voltage-sensitive 45Ca2+ uptake in differentiated NG108-15 cells was
inhibited by nitrendipine, D-600, and diltiazem (IC50 values = 7, 690, and
1600 nM). Our results suggest that VSCCs in neuronal clonal cell lines can
be altered by cellular differentiation. In contrast to those VSCCs involved
in neurotransmitter release, the VSCCs described here appear to be blocked
by organic calcium channel antagonists at very low concentrations.