Characterization of the outward rectifying potassium channel in a novel mouse intestinal smooth muscle cell preparation.
1. The outward rectifying K+ conductance and underlying single channel behaviour in mouse small intestine (MSI) smooth muscle cells was studied using microelectrode impalement and the patch clamp technique. 2. At 37 degrees C, smooth muscle cells in MSI explants had a resting membrane potential around -65 mV and showed spontaneous electrical and mechanical activity. 3. Under whole-cell voltage clamp, depolarization of smooth muscle cells in the explants evoked a methoxyverapamil (D600)-sensitive, partially inactivating inward current and a non-inactivating outward current. The outward current was also observed in enzymatically dispersed cells from neonatal mouse small intestine. 4. The reversal potential of the outward current as established in tail current experiments was -70.2 mV. Tail currents could be fitted with a single exponential, suggesting the participation of only one population of channels. 5. The outward current was sensitive to 4-aminopyridine (10(-4) M), Ba2+ (1 mM) and to the presence of Cs+ in the pipette, but not to D600 (10(-6) M), or the presence of ATP (1 mM) in the pipette. 6. In the cell-attached patch configuration, a unitary outward current was observed that showed increased activity upon depolarization of the patch. The current-voltage relationship was close to linear with a slope conductance of 186 pS. 7. With normal K+ (6 mM) in the pipette, the extrapolated reversal potential for the unitary current was around -75 mV, while with high K+ (120 mM) the reversal potential was close to 0 mV. 8. Averaging single channel traces recorded under a depolarizing pulse protocol resulted in a trace with similar time characteristics as the outward current observed in the whole-cell configuration. 9. The burst behaviour of the channel was described by a simple model consisting of two closed states, Cf (intraburst closed state) and Cs (interburst closed state) and an open state (O). The rate constants in the model showed differential sensitivity to potential changes, channel blockade by Ba2+ and equimolar K+ conditions. 10. It was concluded that the outward rectifying potassium current in MSI smooth muscle cells is mediated by a 186 pS bursting channel. Voltage dependency and Ba2+ blockade are mainly reflected by changes in the transition rate from the open channel state to the interburst closed state.
Molleman
A
A
Department of Biomedical Sciences, McMaster University, Hamilton, Ontario, Canada.
Thuneberg
L
L
Huizinga
J D
JD
D015761:4-Aminopyridine / Q000494:pharmacology; D000818:Animals; D000831:Animals, Newborn; D001464:Barium / Q000494:pharmacology; D004110:Diltiazem / Q000494:pharmacology; D004594:Electrophysiology; D005260:Female; D066298:In Vitro Techniques; D007422:Intestines / Q000166:cytology / Q000502:physiology; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects; D051379:Mice; D008815:Mice, Inbred Strains; D008954:Models, Biological; D009130:Muscle, Smooth / Q000166:cytology / Q000502:physiology*; D009467:Neuromuscular Depolarizing Agents / Q000494:pharmacology; D015221:Potassium Channels / Q000502:physiology*
D009467:Neuromuscular Depolarizing Agents; D015221:Potassium Channels; D001464:Barium; D015761:4-Aminopyridine; D004110:Diltiazem