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Volume 16, Number 15,
Issue of August 1, 1996
pp. 4596-4603
Copyright ©1996 Society for Neuroscience
Involvement of a Phorbol Ester-Insensitive Protein Kinase C in
the  2-Adrenergic Inhibition of Voltage-Gated Calcium
Current in Chick Sympathetic Neurons
Received Feb. 22, 1996; revised April 22, 1996; accepted May 13, 1996.
Stefan Boehm1, 2,
Sigismund Huck1, and
Michael Freissmuth2
Departments of 1 Neuropharmacology and
2 Pharmacology, University of Vienna, A-1090 Vienna,
Austria
 2-Adrenoceptors regulate the
efficacy at the sympatho-effector junction by means of a feedback
inhibition of transmitter release. In chick sympathetic neurons, the
mechanism involves an inhibition of N-type calcium channels, and we now
present evidence that this effect involves an atypical, phorbol
ester-insensitive protein kinase C (PKC). The inhibition of
voltage-gated Ca2+ currents by the specific
2-adrenergic agonist UK 14,304 was
significantly attenuated when the PKC inhibitors PKCI(19-36),
staurosporine, or calphostin C were included in the internal solution
used to fill the patch pipettes, or if staurosporine or calphostin C
were applied extracellularly; however, phorbol esters as classical
activators of PKC or oleoylacetylglycerol did not mimic the effect of
UK 14,304, and chronic exposure to 4- -phorbol dibutyrate (PDBu) did
not attenuate it, even though PKC and - isozymes were
translocated to plasma membranes by PDBu. The atypical isozyme PKC
was translocated by 100 µM arachidonic acid
(AA), but not by PDBu; 100 µM AA and linoleic
acid inhibited voltage-activated Ca2+ currents,
and this effect was attenuated when PKCI(19-36) was added to the patch
pipette solution. Our observations indicate that classical, new, and
atypical PKC isozymes are present in chick sympathetic neurons and that
an atypical, phorbol ester-insensitive PKC is involved in the
inhibition of voltage-activated calcium currents by
2-adrenoceptor activation.
Key words:
2-adrenoceptor;
calcium current;
chick
sympathetic ganglion;
protein kinase C isozymes;
phorbol ester;
arachidonic acid
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