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1. Under voltage-clamped conditions, gastric smooth muscle cells of BALB/c mice developed spontaneous (STOCs) and caffeine- (I(CAF)) and carbachol-induced (I(CCh)) transient outward currents. 2. In fura-2 microscopic measurements of intracellular Ca(2+) concentration ([Ca(2+)](i)), caffeine and carbachol (CCh) provoked similar transient [Ca(2+)](i) elevations. 3. Both I(CCh) and CCh-induced [Ca(2+)](i) elevation of single smooth muscle cells occurred in an 'all-or-nothing' fashion in contrast to the reproducible caffeine responses. 4. On the basis of the suppression of STOCs and I(CAF) by nicardipine, tetraethylammonium and iberiotoxin, but not by charybdotoxin nor apamin, it was suggested that both currents were generated by large conductance type Ca(2+)-activated K(+) channels. 5. In measurements of isometric tension, caffeine produced relaxation of gastric smooth muscle strips in a concentration-dependent manner (0.1 -- 3 mM). The concentration-dependent relaxation with caffeine was mimicked by dibutyryl cyclic AMP which produced potentiation of contraction triggered by 50 mM KCL. 6. At caffeine concentrations >3 mM, a transient contraction followed by relaxation was provoked as the quasi maximal response to caffeine. In the quasi maximal response, caffeine acted as a potent relaxant in smooth muscle strips precontracted with 50 mM KCl or 3 microM CCh. 7. The relaxation with caffeine was significantly accelerated in those strips precontracted with KCl or CCh. All these results suggest that ryanodine-sensitive Ca(2+) release, which is triggered by caffeine, is an important modifier of Ca(2+) homeostasis in the cytoplasm and the contractility of gastric smooth muscle cells of mice.

1. Effects of penicillin G (PCN) on glycine (Gly)-evoked Cl- current (IGly) were investigated in acutely dissociated rat ventro-medial hypothalamic (VMH) neurones by the whole cell mode of patch clamp technique. 2. When PCN was applied simultaneously with Gly, PCN depressed IGly like a Cl- channel blocker. 3. The PCN-induced blocking action was clearly observed at a low PCN concentration (30 u), while the maximal blockade was achieved by 600 u (units per 10 ml) PCN. 4. When tested solution containing both PCN and Gly was quickly substituted with one containing Gly only, a new rebound-like transient current (I(T)) which also passed through Cl- channel, was elicited. 5. The peak amplitude of I(T) induced by PCN at concentrations higher than 100 u was greater than that induced by glycine alone. We termed this phenomenon PCN-induced potentiation of IGly. In all cells tested, PCN potentiated IGly. 6. At a lower PCN concentration below 30 u, I(T) generation was not clear in the presence of 10(-5) M gamma-aminobutyric acid. With PCN a higher concentration than 300 u, I(T) amplitude was greater than that of the original peak IGly. This was observed in 18 neurones out of 21. The maximal amplitude of the I(T) was achieved with 600 u PCN.

1. The contribution of Cl- redistribution to the decay phase of the GABA (gamma-aminobutyric acid) response was investigated in isolated frog sensory neurones, using a suction-pipette technique which allows for internal perfusion under conditions of voltage clamp. 2. In neurones perfused with 120 mM [Cl-]i and [Cl-]o at driving forces (delta VH) of less than 15 mV, no shift of GABA equilibrium potential (EGABA) occurred during a continuous application of GABA, at various concentrations. However, increases of delta VH towards negative or positive potentials over 15 mV induced EGABA shifts. 3. The degree of EGABA shift was governed by the total amount of Cl- flux across the soma membrane, an event which depends upon delta VH, GABA concentration and drug application time. 4. The time-dependent EGABA shift due to Cl- redistribution during GABA application induced a current run-down resulting from a decreased Cl- gradient and a diminished Cl- conductance (gCl), the latter brought about by a drop in the intracellular ionic density of Cl-. 5. The EGABA shift during a continuous GABA application was also affected by [Cl-]i; e.g. the shift more readily occurred at lower [Cl-]i. 6. In neurones perfused with internal and external solutions containing 120 mM-Cl- at a delta VH of less than 10 mV, the change of gCl occurred with no shift of EGABA during the continuous application of GABA at concentrations over 6 x 10(-5) M, thereby indicating a 'real' GABA receptor desensitization. The desensitization depended solely upon the agonist concentrations but not upon the amount of ICl. Under these conditions, the time course of recovery from GABA desensitization was estimated. The decrease of gCl at the desensitization phase was a single exponential. 7. At a delta VH greater than 15 mV, therefore, the decay of ICl induced by GABA concentrations over 6 x 10(-6) M consists of the sum of both the 'real' GABA receptor desensitization and the current run-down brought about by Cl- shifts. The gCl at the current decay phase consisted of a double exponential. In the present experiments we chose experimental conditions with which Cl- shift become negligible. 8. The 'pure' GABA receptor desensitization during a continuous application of GABA developed rapidly at GABA concentrations over 10(-5) M. The speed of desensitization was facilitated by increasing the magnitude of desensitization.

1. The kinetic properties of the activation and inactivation (desensitization) phases of pentobarbitone (PB)-induced inward Cl- current (ICl) were studied in isolated frog sensory neurones, following suppression of Na+, K+ and Ca2+ currents, using the concentration jump technique which combines the internal perfusion and the rapid exchange of the external solutions surrounding a neurone with time constants of 2-3 ms. The results were compared with those of the gamma-aminobutyric acid (GABA)-gated ICl. 2. The PB dose-response curve was bell-shaped and the maximum peak value was less than the current induced by 1.7 X 1.5(-5) M-GABA, the concentration at which GABA evoked a half-maximum response. 3. The activation and inactivation phases of PB-induced ICl consisted of double-exponential, fast and slow components, respectively. The time constant of the fast component (tau af) of the activation was relatively stable in a concentration range between 3 X 10(-4) and 6 X 10(-3) M. The time constant of the slow component (tau as) of the activation decreased with increasing PB concentrations. Both the fast and slow components (tau if and tau is) of the inactivation decreased with increasing PB concentrations. 4. Over a wide range of concentrations the tau af and tau as values of the PB-induced ICl were 10-30 times greater than the respective values of GABA-induced ICl. 5. At concentrations below 10(-3) M the PB-induced ICl was voltage dependent at more negative potentials than -20 mV. 6. The PB-induced ICl was blocked by bicuculline and by picrotoxin, but in a different manner. Bicuculline increased the time constants of the activation and inactivation. Picrotoxin had little effect on the activation phase but markedly facilitated the inactivation phase. 7. High concentrations of PB (over 10(-3) M) led to a decline in both the peak and plateau currents of the PB-induced ICl. A transient 'hump' current appeared with wash-out of the external solutions containing high concentrations of PB. This hump current was blocked by bicuculline in a dose-dependent manner. 8. The results suggest the possibilities that the PB receptor-ionophore complexes consist of at least two different components having different affinities and kinetics and that the PB and GABA binding sites are closely located.