Effects of intracellular K+ and Rb+ on gating of embryonic rat telencephalon Ca(2+)-activated K+ channels.
We have investigated the effects of intracellular K+ and Rb+ on single-channel currents recorded from the large-conductance Ca(2+)-activated K+ (BK) channel of the embryonic rat telencephalon using the inside-out patch-clamp technique. Our novel observation concerns the effects of these ions on rapid flickering of channel openings. Specifically, flicker gating was voltage dependent, i.e., it was reduced by depolarization in the -60 to -10 mV range with equimolar concentrations of K+ ions (150 Ko+/150 Ki+). Removal of Ki+ resulted in significant flickering at all potentials in this voltage range. In other words, the voltage dependence of flicker gating was effectively eliminated by the removal of Ki+. This suggests that a K+ ion entering the channel from the intracellular medium binds, in a voltage-dependent manner, at a site that locks the flicker gate in its open position. No effects of changes in Ki+ were observed on the primary, voltage-dependent gate of the channel. The change in flickering did not cause a change in the mean burst duration, which indicates that the primary gate is stochastically independent of the flicker gate. Intracellular Rb+ can substitute for--and is even more effective than--Ki+ with regard to suppression of flickering. Substitution of Rbi+ for Ki+ also increased the mean burst duration for V > or = -30 mV. Both effects of Rbi+ were removed by membrane hyperpolarization.
Mienville
J M
JM
Laboratory of Neurophysiology, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland 20892, USA. jmm@zippy.nih.gov
Clay
J R
JR
D000818:Animals; D055592:Biophysical Phenomena; D001703:Biophysics; D002118:Calcium / Q000378:metabolism*; D004622:Embryo, Mammalian / Q000378:metabolism; D066298:In Vitro Techniques; D007424:Intracellular Fluid / Q000378:metabolism; D015640:Ion Channel Gating; D007700:Kinetics; D008564:Membrane Potentials; D008954:Models, Biological; D011188:Potassium / Q000378:metabolism* / Q000494:pharmacology; D015221:Potassium Channels / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D012413:Rubidium / Q000378:metabolism* / Q000494:pharmacology; D013687:Telencephalon / Q000378:metabolism
D015221:Potassium Channels; D012413:Rubidium; D011188:Potassium; D002118:Calcium