1 Smooth muscle cells of the rat portal vein were dispersed by enzymatic treatment and recordings of whole-cell membrane potassium currents were made by the voltage-clamp technique. In isolated cells by use of combined voltage-and current-clamp the effect of BRL 38227 on membrane potential and ionic currents was also studied.2 BRL 38227 (0.1 to 10 JM) induced a non-inactivating potassium current (IKCO) which developed slowly (900 s to 300 s, respectively) to its full size. These effects of BRL 38227 were reversible. 3 In addition to its K-channel opening properties, BRL 38227 (1 to 10 JIM) inhibited the amplitude and changed the activation and inactivation characteristics of a slowly-inactivating, calcium influxindependent, outward potassium current (ITO). 6 The induction of IKCO by BRL38227 and the associated hyperpolarization were suppressed by glibenclamide (1 to 10 JIM) in a concentration-dependent manner. Glibenclamide (1 JIM) had no effect on the inhibition of ITO by BRL 38227 (1 JIM).

SUMMARY1. Smooth muscle cells, enzymatically isolated from the antrum of the guinea-pig stomach, were voltage clamped at room temperature using the whole-cell patch clamp technique. In physiological salt solution (PSS), step depolarization from a holding potential of -90 mV elicited inward calcium current (ICa) followed and superimposed by outward potassium current.2. Outward current was divided into components depending on the presence of extracellular Ca2+ and others which were not activated as a result of Ca2+ influx.Ca2+-dependent components were (1) 3. It. activated with a threshold around -30 mV, was fully available at -90 mV and completely inactivated at -10 mV. The time course of both activation and inactivation of Ito at different potentials could be described by single exponential functions. Time constants of activation decreased from 35 ms at -10 mV to 10 ms at + 40 mV. The time constant of inactivation was about 2 s and only weakly voltage dependent. Time constants for exponentially developing recovery from inactivation

1 The effects of levcromakalim and of adenosine 5'-triphosphate (ATP) depletion on membrane potential and ionic currents were studied in freshly-dispersed smooth muscle cells of rat portal vein by use of combined voltage-and current-clamp techniques. 9 The simultaneous inhibition of the delayed rectifier current (ITO) by both levcromakalim and during the development of Ime, is highly significant. It suggests that levcromakalim could modify the interaction of ATP with sites linked to more than one type of K-channel. This results in the opening of those channels which underlie IKCO (and which are normally inhibited by ATP binding) together with the modulation of phosphorylation-dependent channels such as those which underlie ITO.

1. Single smooth muscle cells from the fundus region of the guinea-pig stomach, which showed contractile responses to acetylcholine (ACh) at concentrations greater than or equal to 10(-7) mol/l, were obtained by enzymatic digestion using highly purified collagenase and papain. They were studied by recording membrane currents under voltage clamp with the patch pipette technique in the whole-cell configuration at 25-28 degrees C. 2. By applying voltage jumps from negative holding levels (-70 to -60 mV) to more positive levels, we identified two major activating currents: an initial inward Ca2+ current (ICa) was followed, and partly overlapped, by an outward K+ current (IK). 3. Cholinergic effects on membrane currents were investigated in the range of negative membrane potentials by determining current-voltage relations in the absence of ACh and during its continuous presence in the bathing fluid. 4. ACh induced a decrease in the steady-state conductance which was reversibly blocked by atropine. At physiological external K+ concentration [( K+]o = 6 mmol/l), the reversal potential (Erev) of the current suppressed by ACh (3 x 10(-6) mol/l) was about 20 mV more positive than the calculated K+ equilibrium potential (EK). 5. When [K+]o was increased, Erev was shifted positively; but at each [K+]o, Erev was more positive than EK. 6. Like ACh (10(-6) mol/l), tetraethylammonium (TEA, 1 mmol/l) also suppressed a current with a reversal potential that was, at physiological [K+]o, 20 mV more positive than EK. ACh (10(-5) mol/l) applied in the presence of 1 mmol/l TEA suppressed a pure K+ current (Erev = EK), which was also suppressed by 10 mmol/l TEA. 7. When K+ in the pipette and in the bathing solution was completely replaced by Na+, both ACh (10(-5) mol/l) and TEA (1 mmol/l) caused a reduction of the membrane conductance that appeared to be identical. TEA added to the bathing solution in the presence of ACh did not produce a significant additional conductance decrease. These results did not depend on whether Cl- was present as a charge carrier or not. 8. It is concluded that in fundus muscle of the guinea-pig stomach a major mechanism underlying muscarinic activation is a decrease of a K+ conductance. In addition the results indicate a suppression of a small Na+ conductance which is made up by a population of channels that are also blocked by TEA.

Using the single electrode voltage-clamp technique, vascular smooth muscle cells from the guinea pig portal vein showed an initial inward (Ca2+) current followed by an outward current which peaked within 100 ms and then declined to a steady level in a few seconds. Caffeine (1 mmol/l) selectively blocked the transient component of the outward current (ITO) and allowed differentiation of the outward current into two components: ITO and a caffeine resistant background current. The potassium channel blockers TEA (10 mmol/l) and 4-AP (5 mmol/l) produced about 90% suppression of ITO. ITO was identified as a calcium independent potassium current. Analysis of the inactivation and recovery from inactivation of ITO revealed similarities to the A-current first described for molluscan neurones (11) and later for crista terminalis of rabbit heart (4). Being much slower than this current it also bears similarities in its inactivation kinetics to a transient outward current identified in rabbit portal vein (2, 6).