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Volume 16, Number 21,
Issue of November 1, 1996
pp. 6657-6664
Copyright ©1996 Society for Neuroscience
Modulation of Voltage-Gated Calcium Channels by Orphanin FQ in
Freshly Dissociated Hippocampal Neurons
Received June 5, 1996; revised Aug. 6, 1996; accepted Aug. 9, 1996.
Frederic Knoflach,
Rainer K. Reinscheid,
Olivier Civelli, and
John A. Kemp
F. Hoffmann-La Roche Ltd., Pharma Division, Preclinical Research,
CH-4070 Basel, Switzerland
Orphanin FQ (OFQ) has recently been reported to be an endogenous
ligand for the opioid-like LC132 receptor. The effect of OFQ on high
voltage-gated calcium channels (VGCCs) was examined in freshly
dissociated rat pyramidal neurons using the whole-cell configuration of
the patch-clamp technique. High-threshold Ba2+ currents
were reversibly inhibited by OFQ. The depression of the currents was
associated with a slowed rate of activation and a change in the
activation I-V relationship at step
potentials higher than +30 mV. In concentration-response experiments,
a mean (±SEM) pEC50 value of 7.0 ± 0.07 and a Hill
coefficient of 1.5 ± 0.08 (n = 5) were
obtained. The near-maximum inhibition of the Ba2+ currents
by OFQ (1 µM) amounted to 31 ± 2.2% of control
(n = 15). Opioid receptors could not account for
the effects of OFQ on VGCCs, because naloxone, a broad spectrum µ-,
 -, and  -receptor antagonist, did not reduce the effectiveness of
OFQ. When GTP- -S was included in the pipette, the depression of the
currents by OFQ was irreversible, whereas currents from neurons
preincubated with pertussis toxin were not inhibited by OFQ, consistent
with the involvement of a PTX-sensitive G-protein. When selective
blockers of VGCCs were used, it was demonstrated that all subtypes of
VGCCs were affected by OFQ. In conclusion, the effect of OFQ on VGCCs
expressed in hippocampal CA3 and CA1 neurons may play an important role
in the regulation of hippocampal cell excitability and neurotransmitter
release.
Key words:
calcium channel blockers;
calcium channel drug
effects;
opioid drug effects;
G-protein;
patch clamp;
nifedipine;
 -conotoxin-GVIA;
-agatoxin-IVA;
GTP--S;
orphanin FQ;
nociceptin;
opioid receptor;
neuromodulation
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