M. Fejtl et al., MERCURIC(II) CHLORIDE MODULATES SINGLE-CHANNEL PROPERTIES OF CARBACHOL-ACTIVATED CL- CHANNELS IN CULTURED NEURONS OF APLYSIA-CALIFORNICA, Cellular and molecular neurobiology, 14(6), 1994, pp. 665-674
1. The effect of mercuric(II) chloride on kinetic parameters of carbac
hol-activated single chloride channels were studied in cultured neuron
s of the marine mollusk, Aplysia californica. 2. Single neurons of Apl
ysia were cultured in L-15 medium containing 1 mM beta-D-xyloside, whi
ch improved the success rate for gigaseal formation by 46%. Carbachol-
activated single chloride channels were recorded in the cell-attached
patch clamp configuration. Recordings with control solution (1 mu M ca
rbachol) and with test solution (1 mu M carbachol + 1 mu M HgCl2) were
performed successively on the same neuron. 3. In both the control and
the test solution the open and closed time distributions were fitted
with a double-exponential function. However, kinetic analysis revealed
that Hg2+ caused a significant reduction of the mean closed time (10.
37 +/- 1.08 vs. 3.32 +/- 0.02 msec) and of the second time constant ta
u(2) of the dosed time distribution (2.09 +/- 0.05 VS. 0.66 +/- 0.5 ms
ec). The reduction of tau(2), i.e., fewer events in the longer closed
state under the action of Hg2+, may be the physical cause for the redu
ction of the mean closed time and thus underlies the increased open pr
obability p(0) (0.13 +/- 0.01 vs. 0.29 +/- 0.01 msec) of carbachol-act
ivated chloride channels. 4. Inorganic Hg2+ affects the acetylcholine
receptor at lower concentrations than previously reported.