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Journal of Neuroscience, Vol 13, 3884-3894, Copyright © 1993 by Society for Neuroscience
Kinetic basis for the voltage-dependent inhibition of N-type calcium current by somatostatin and norepinephrine in chick sympathetic neurons
A Golard and SA Siegelbaum
Department of Pharmacology, Howard Hughes Medical Institute, Columbia University, New York, New York 10032.
Neurotransmitter inhibition of calcium currents (ICa) can be relieved by
large depolarizing prepulses. This effect has been postulated to be due
either to the voltage-dependent unbinding of an inhibitory molecule from
the channel or to a slow voltage-dependent gating step intrinsic to the
modulated channel. According to the first hypothesis, the rate of
reinhibition (reblock) following a depolarizing prepulse should depend on
the concentration of active inhibitory molecules and thus should increase
with the extent of inhibition. To distinguish between these models we
examined the actions of norepinephrine (NE) and somatostatin (SS) on
high-threshold calcium currents in chick sympathetic ganglia, using
whole-cell voltage-clamp methods. As previously described in other systems,
both NE and SS inhibit omega- conotoxin-sensitive N-type Ca2+ current in a
voltage-dependent manner. Pertussis toxin (PTX) pretreatment prevents the
inhibition of the current, while replacing GTP in the patch pipette with
GTP-gamma-S results in irreversible inhibition, consistent with the
involvement of a PTX-sensitive G-protein. The inhibitory responses to NE
and SS are not additive, suggesting that they act at a common locus. The
inhibitory response to repeated applications of NE or SS desensitizes, with
little evidence for cross desensitization. The inhibition of ICa is
relieved by a 15 msec prepulse to +100 mV. Following repolarization to -80
mV, ICa slowly reblocks. During prolonged applications of NE or SS the
extent of inhibition decreases due to desensitization and reblock kinetics
are significantly slowed (time constant increases from 60 msec to > 100
msec for both NE and SS). These results are well fit by a quantitative
model in which the kinetics of reblock reflect the binding of an inhibitory
molecule to the channel.
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