Distinct modes of channel gating underlie inactivation of somatic K+ current in rat hippocampal pyramidal cells in vitro.
1. We have used the cell-attached configuration of the patch-clamp recording method to characterize the biophysical properties of the voltage-gated K+ channel underlying a 4-aminopyridine (4-AP)- and tetraethylammonium (TEA)-sensitive K+ current (IK(AT)) in pyramidal cells of hippocampal slice cultures. 2. The unitary conductance of channels carrying IK(AT) current (KAT channels) was 19.1 +/- 5.1 pS with a physiological K+ gradient (2.7 mM external K+) and 39.0 +/- 3.6 pS with high external K+ (140 mM). The reversal potential changed with the external K+ concentration as expected for a channel with a dominant K+ selectivity. Channel activity was blocked under both conditions by either external application of 4-AP at 100 microM or by including 20 mM TEA in the pipette solution. 3. An analysis of kinetic behaviour showed that open times were distributed as a single exponential. The mean open time (+/- S.D.) was 4.4 +/- 1.4 ms at a voltage 30 mV positive to resting potential and increased with further depolarization to reach a value of 16.2 +/- 7.4 ms at 70 mV positive to the resting potential. At this depolarized potential, we observed bursts of channel openings with a mean burst duration around 100 ms. 4. With repeated depolarizing pulses, response failures of the KAT channel occurred in a non-random manner and were grouped (referred to as mode 0). This mode was associated with a voltage-dependent inactivation process of the channel and was favoured when the opening probability of the channel was reduced by increasing steady-state inactivation or by bath application of 4-AP. This is consistent with the localization of the binding site for 4-AP at or near the inactivation gate of the channel. 5. When KAT channel openings were elicited by 500 ms depolarizing steps, activity was either transient or it persisted throughout the duration of the pulse. These two modes of activity alternated in a random manner or occurred in groups giving rise to transient (time constant, 20-100 ms) or sustained ensemble currents. In the presence of low concentrations of 4-AP (20-40 microM), the transient pattern of activity was more frequently observed. 6. In addition to mode 0, we propose the existence of at least two further gating modes for KAT channels: mode T (transient current) and mode S (sustained current) that underlie the three decaying components of the IK(AT) ensemble current. These gating modes are probably under the control of intracellular factors that remain to be identified.
Bossu
J L
JL
Brain Research Institute, University of Zurich, Switzerland. jlbossu@neurochem.u-strabg.fr
Gähwiler
B H
BH
D015761:4-Aminopyridine / Q000494:pharmacology; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D046508:Culture Techniques; D004594:Electrophysiology; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D015640:Ion Channel Gating / Q000187:drug effects / Q000502:physiology*; D007700:Kinetics; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D018408:Patch-Clamp Techniques; D015221:Potassium Channels / Q000187:drug effects / Q000502:physiology*; D017966:Pyramidal Cells / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D051381:Rats; D013757:Tetraethylammonium Compounds / Q000494:pharmacology
D015221:Potassium Channels; D013757:Tetraethylammonium Compounds; D015761:4-Aminopyridine