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Vol. 291, Issue 2, 893-902, November 1999
Department of Physiology and Pharmacology, Center for the
Neurobiological Investigation of Drug Abuse, Wake Forest University
School of Medicine, Winston-Salem, North Carolina
Cannabinoid (CB1) receptor activation produced
differential effects on voltage-gated outward potassium currents in
whole-cell recordings from cultured (7-15 days) rat hippocampal
neurons. Voltage-dependent potassium currents A (IA) and D
(ID) were isolated from a composite
tetraethylammonium-insensitive current (Icomp) by blockade
with either 4-aminopyridine (500 µM) or dendrotoxin (2 µM) and
subtraction of the residual IA from Icomp to
reveal ID. The time constants of inactivation (
) of
IA and ID as determined in this manner were
found to be quite different. The CB1 agonist WIN 55,212-2 produced a 15- to 20-mV positive shift in voltage-dependent inactivation of IA and a simultaneous voltage-independent
reduction in the amplitude of ID in the same neurons. The
EC50 value for the effect of WIN 55,212-2 on ID
amplitude (13.9 nM) was slightly lower than the EC50 value
for its effect on IA voltage dependence (20.6 nM).
Pretreatment with either the CB1 antagonist SR141716A or
pertussis toxin completely blocked the differential effects of WIN
55,212-2 on IA and ID, whereas cellular
dialysis with guanosine-5'-O-(3-thio)triphosphate mimicked the action of cannabinoids but blocked the action of simultaneously administered cannabinoid receptor ligands. Finally, the
differential effects of cannabinoids on IA and
ID were both shown to be mediated via the well documented
cannabinoid receptor inhibition of adenylyl cyclase and subsequent
modulation of cAMP and protein kinase. These actions are considered in
terms of cAMP-mediated phosphorylation of separate IA and
ID channels and the contribution of each to composite
voltage-gated potassium currents in these cells.
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