Extracellular bupivacaine inhibits GIRK1/4 channels. Oocytes were
injected with the cRNA for GIRK4-GIRK1 dimer plus the m2 muscarinic
receptor. (a) Macroscopic current was recorded
continuously by using two-electrode voltage clamp from an oocyte bathed
in 95 mM KCl (pH 7.5) with the muscarinic agonist carbachol (0.3 μM;
solid bar) and then with carbachol plus 500 μM bupivacaine (gray
box). The holding potential (VH) was −80
mV. Dashed line indicates the zero current level.
(Right) Coapplication of carbachol and bupivacaine (500
μM). (b) Continuous current recording shows the effect
of increasing concentrations of bupivacaine coapplied with carbachol.
(c) The carbachol-induced current was normalized
(carbachol, I/Io) and
plotted as a function of bupivacaine concentration. Smooth curve shows
the best fit to the Hill equation, with an apparent
Ki of 22 ± 4 μM and a Hill
coefficient of 0.68 ± 0.06 (n = 7).
(d) Xenopus oocytes injected with the
cRNA for GIRK1, GIRK4, and the G protein Gβ1
and Gγ2 subunits. Current responses were
elicited by 500-ms voltage step to −80 mV
(VH = 0 mV) in 0, 30, 100, or 500 μM
bupivacaine. (e) The normalized current
(Gβγ,
I/Io) was plotted as a
function of bupivacaine concentration. Smooth curves show the best fit
to the Hill equation for −40 mV (Ki =
170 ± 12 μM; Hill = 0.83 ± 0.04) and −110 mV
(Ki = 98 ± 3 μM; Hill =
0.96 ± 0.03).