Values Of Membrane Potential Research Articles

Attenuation of endothelium-dependent hyperpolarizing factor by bacterial lipopolysaccharides

Endothelium-dependent hyperpolarizing factor (EDHF) is an important contributor to agonist-induced vascular dilation. Recent studies suggest that bacterial lipopolysaccharides attenuate endothelium-dependent dilation. Whether or not this effect is mediated through inhibition of EDHF is not known. We studied the in vitro influence of Escherichia coli lipopolysaccharides on endothelium-dependent smooth muscle dilation and hyperpolarization in porcine coronary arteries. Endothelium-intact porcine coronary arterial rings were examined after 20 h of incubation with either saline or E. coli lipopolysaccharides (100 μg/ml). Endothelium-dependent dilation elicited by increasing concentrations of bradykinin was significantly attenuated by lipopolysaccharides. Baseline values of smooth muscle membrane potential were not influenced by lipopolysaccharides. However, lipopolysaccharides significantly attenuated bradykinin-induced smooth muscle membrane hyperpolarization. Our results suggest that attenuation of EDHF is an important mechanism through which lipopolysaccharides influence vascular dilation in severe sepsis.

Effect of 1-methyl-4-phenylpyridinium (MPP+) on mitochondrial membrane potential in cerebellar neurons: interaction with the NMDA receptor.

The effect of MPP+, a dopaminergic neurotoxin, in mitochondrial membrane potential was investigated in dissociated cerebellar granule cells using rhodamine 123 and flow cytometry. MPP+ (1 mM) decreased the mitochondrial membrane potential by 30%. Antagonists of the NMDA receptor complex, such as MK-801 (IC50 value of 20.92 +/- 0.02 nM), 5,7-dichlorokynurenic acid (IC50 value of 6.46 +/- 1.06 microM) and D-AP5 (IC50 value of 8.29 +/- 0.63 microM), inhibited the action of MPP+. Neither NBQX, nor riluzole, nor desipramine modified the action of MPP+. Dibucaine restored the basal values of mitochondrial membrane potential altered by MPP+. Since, in the presence of NMDA, MPP+ antagonized the effect of this total agonist, it can be concluded that, in this preparation, MPP+ interacts with the NMDA receptor complex as a partial agonist. This interaction could be the result of an allosteric modulation of the NMDA receptor complex by MPP+. The decrease of mitochondrial membrane potential induced by MPP+ is antagonized by dibucaine, suggesting that this effect is mediated by an activation of phospholipase A2.

Effect of Adsorption of Charged Macromolecules on Streaming and Membrane Potential Values Measured with a Microporous Polysulfone Membrane

Changes in streaming and membrane potentials measured across a commercial microporous polysulfone membrane as a result of the adsorption of differently charged macromolecules were studied. Measurements were carried out with different NaCl solutions (10−3 M to 5 × 10−2M) and their mixtures with a polyanion (dextran sulfate or DS) and a polycation (diethylaminoethyl or DEAE-dextran). From electrokinetic and electrochemical measurements, information about characteristic membrane parameters (transport number and ionic permselectivity) and membrane/solution interactions (zeta potential) can be obtained. Results show how the adsorption of charged particles affects the electrical parameters of the membrane and also its hydraulic permeability. The concentration dependence of these parameters was also studied.

VITALITY OF CIDER YEAST GROWN MICRO-AEROBICALLY WITH ADDED ETHANOL, BUTAN-J-OL ORISO-BUTANOL

Vitality and viability of an alcohol-tolerant wine yeast, used in cider production, were assessed after exposure to alkanol(s) during growth. Criteria employed were: methylene blue reduction, ability to form colonies on yeast extract-peptone-glucose agar medium, glucose driven proton efflux rates (“acidification power”), fermentative (CO2 output) rates and adenylate energy charge values. We also monitored the maintenance of transmembrane electrochemical potential across the plasma membrane as measured by flow cytometry and by scanning confocal laser microscopy of oxonol dye exclusion. Growth rates were diminished by a third by 7.5% (v/v) added ethanol, 1% butan-1-ol or 1.4% iso-butanol. Exposure to 10% (v/v) ethanol gave 16% loss of “viability”, as measured by methylene blue reduction, during the first 20 h of growth. For 1% butan-1-ol, 50% loss of “viability” occurred over 40 h, whereas a similar effect of iso-butanol took 55 h. Adenylate charge values were high (<0.8) in growing cultures, remained high in early stationary phase but declined to 0.4 after 115 h. These values were hardly affected by 5 or 7.5% (v/v) ethanol whereas 10% or 15% (v/v) ethanol gave values of 0.58 and 0.16 after only 5 h exposure. 1% butan-1-ol or iso-butanol decreased adenylate charge values to a greater extent than 10% (v/v) ethanol, with the straight chain alcohol the more potent. Oxonol exclusion indicated that the vast majority of colls with greatly diminished vitality have maintained the plasma membrane potential values required to retain viability, despite extensive exposure to alkanol(s). Thus loss of ability to reduce methylene blue indicates diminished vitality but is not a reliable index of loss of viability. “Acidification power” was a more sensitive indicator of vitality than adenylate charge values. When mixtures of C2 + C4 alcohols were employed effects were generally additive rather than synergistic.

Effect of unstirred solution layers on the thermal membrane potential through cation-exchange membranes

The thermal membrane potential through a cation-exchange membrane has been determined under different experimental situations. In all cases, a good linear dependence between the thermal membrane potential, Δψ, and the temperature difference, ΔT, established between both external electrolyte solutions has been observed. The value of Δψ decreases notably when the concentration of the electrolyte solution used is increased and it also depends on the type of the cation involved, so that Δψ decreases when the corresponding atomic number increases. The value of the temperature coefficient, Δψ ΔT , is notably affected by the unstirred solution layers adjoining the membrane surfaces. A theoretical model is presented to determine the influence of the temperature polarization effect on the thermal membrane potential value.

Two types of low-voltage-activated Ca2+ channels in neurones of rat laterodorsal thalamic nucleus.

1. The pharmacological and kinetic properties of two types of low-voltage-activated (LVA) Ca2+ currents were studied in thalamocortical neurones of the laterodorsal (LD) thalamic nucleus during early postnatal development. The whole-cell patch-clamp technique was used on brain slices from rats of three age groups: 12, 14 and 17 days old (postnatal day (P) 12, P14 and P17). 2. In P12 neurones, the population of LVA Ca2+ channels was homogeneous. LVA Ca2+ current elicited by depolarizing voltage steps from a holding potential more negative than -70 mV was sensitive to nifedipine (Kd = 2.6 microM). This current reached a maximum at about -55 mV and had a fast monoexponential decay with a time constant, tau h,f, of 32.3 +/- 4.0 ms. 3. The population of LVA Ca2+ channels in P14 and P17 neurones was found to be heterogeneous. A subpopulation of nifedipine-insensitive LVA Ca2+ channels was observed. The current-voltage curve of the Ca2+ current had a characteristic hump with two peaks at about -65 and -55 mV. As well as the fast component (designated IT,f), the decay of the LVA current also included a slow component (designated IT,s), with inactivation time constants (tau h,s) of 54.2 +/- 4.5 and 68.6 +/- 3.17 ms for P14 and P17 neurones, respectively. 4. The kinetics of both components could be well approximated by the m2h Hodgkin-Huxley equation. No significant difference in activation kinetics was observed. The activation time constants for the fast (tau m,f) and slow (tau m,s) components were 6.3 +/- 1.0 and 7.3 +/- 1.5 ms, respectively. 5. La3+ at a concentration of 1 microM effectively blocked the IT,f component but Ni2+ (25 microM) completely eliminated the IT,s component. 6. Steady-state inactivation curves of both components could be best fitted by a Boltzmann function with membrane potential values at half-maximal inactivation of -85.5 and -98.1 mV for the fast and slow components, respectively. 7. It was concluded that two different subtypes of LVA Ca2+ channel are present in LD neurones. Only the fast type is well expressed at the earliest postnatal stage (P12). The slow type could be found at the end of the second week (P14). The amplitude of the slow current increased progressively up to P17, obviously coinciding with dendritic expansion as judged by progressive increase of the membrane capacitance of the corresponding neurones. This property appears to differentiate neurones of the associative nuclei from neurones of other thalamic nuclei.

Human red blood cell membrane potential and fluidity in glucose solutions

This study aimed to investigate the membrane potential and fluidity changes of human red blood cells subjected to isotonic glucose solutions. For the control erythrocytes a membrane potential value of -10.1 ± 1.8 mV was obtained, whereas erythrocyte membranes in diabetic cells were hyperpolarized, with a potential of -13.9±2.3 mV. Incubation of both types of red blood cells with increasing glucose concentrations resulted in a substantial hyperpolarization of the cell membranes. Glucose solutions in water had a much stronger effect on membrane potential than glucose dissolved in phosphate-buffered saline. Red blood cell membrane fluidity measurements using the fluorescent label TMA-DPH did not reveal any significant changes upon incubation with glucose.

Probing excitation-contraction coupling in trachealis smooth muscle with the mycotoxin cyclopiazonic acid.

1. Muscarinic stimulation-induced tonic contraction of airway smooth muscle is independent of membrane potential. This contraction is not sensitive to inhibition by voltage-operated Ca2+ channel blockers or by K+ channel openers. 2. Cyclopiazonic acid (CPA) inhibits Ca2+ loading of internal stores but does not affect maximal tonic contraction induced by acetylcholine (ACh) in steady state conditions. 3. After depletion of internal Ca2+ stores with CPA, ACh-induced tonic contraction becomes dependent upon values of membrane potential. The contraction is then sensitive to voltage-operated Ca2+ channel blockers and to K+ channel openers. 4. Treatment of trachealis muscle with CPA potentiates the M2-mediated component of ACh stimulation, but this potentiation is not entirely responsible for the switch in excitation-contraction (E-C) coupling. 5. It is proposed that depletion of internal Ca2+ stores with CPA and promotion of M2-stimulation can lead to a switch in E-C coupling in trachealis smooth muscle from pharmaco- to electromechanical mode, perhaps by targeting a plasma membrane K+ channel.

Top-down control analysis of temperature effect on oxidative phosphorylation

The effects of temperature on the control of respiration rate, phosphorylation rate, proton leakage rate, the protonmotive force and the effective ATP/O ratio were determined in isolated rat liver mitochondria over a range of respiratory conditions by applying top-down elasticity and control analyses. Simultaneous measurements of membrane potential, oxidation and phosphorylation rates were performed under various ATP turnover rates, ranging from state 4 to state 3. Although the activities of the three subsystems decreased with temperature (over 30-fold between 37 and 4 degrees C), the effective ATP/O ratio exhibited a maximum at 25 degrees C, far below the physiological value. Top-down elasticity analysis revealed that maximal membrane potential was maintained over the range of temperature studied, and that the proton leakage rate was considerably reduced at 4 degrees C. These results definitely rule out a possible uncoupling of mitochondria at low temperature. At 4 degrees C, the decrease in ATP/O ratio is explained by the relative decrease in phosphorylation processes revealed by the decrease in depolarization after ADP addition [Diolez and Moreau (1985) Biochim. Biophys. Acta 806, 56-63]. The change in depolarization between 37 and 25 degrees C was too small to explain the decrease in ATP/O ratio. This result is best explained by the changes in the elasticity of proton leakage to membrane potential between 37 and 25 degrees C, leading to a higher leak rate at 37 degrees C for the same value of membrane potential. Top-down control analysis showed that despite the important changes in activities of the three subsystems between 37 and 25 degrees C, the patterns of the control distribution are very similar. However, a different pattern was obtained at 4 degrees C under all phosphorylating conditions. Surprisingly, control by the proton leakage subsystem was almost unchanged, although both control patterns by substrate oxidation and phosphorylation subsystems were affected at 4 degrees C. In comparison with results for 25 and 37 degrees C, at 4 degrees C there was evidence for increased control by the phosphorylation subsystem over both fluxes of oxidation and phosphorylation as well as on the ATP/O ratio when the system is close to state 3. However, the pattern of control coefficients as a function of mitochondrial activity also showed enhanced control exerted by the substrate oxidation subsystem under all intermediate conditions. These results suggest that passive membrane permeability to protons is not involved in the effect of temperature on the control of oxidative phosphorylation.

Membrane potential of smooth muscle cells of human allantochorial placental vessels

The membrane potential of vessel endothelial and smooth muscle cells is central to their function of regulating blood flow. In the present study, the membrane potential (MP) of smooth muscle cells of human placental arteries and veins with or without endothelium is observed. MP is classically obtained using microelectrodes inserted into the smooth muscle cells, either through the endothelium or directly. MP was found to remain stable during 120–140 min and was independent of pH over a wide physiological range. Variation of external temperature induced depolarization at 20°C. Variations in external concentrations of Na + and K + did not influence MP values, while increases in Ca 2+ and Mg 2+ concentrations caused depolarization. The membranes of vascular smooth muscle cells were also depolarized by ouabain. These data suggest that endothelial cell exert a protective effect (release of constricting or relaxing factors) and that MP is regulated, in particular, by lipid-calcium interaction, opening of voltage-dependent Ca 2+-channels and by ATPase.

Functional Heterogeneity of an Isolated Mitochondrial Population Revealed by Cytofluorometric Analysis at the Single Organelle Level

Isolated rat liver mitochondria were incubated under various metabolic conditions to determine their membrane potential (MMP) as measured continuously by a tetraphenylphosphonium (TPP+)-selective electrode. By flow cytometry, a parallel analysis of fluorescence emissions observing single mitochondria stained with the lipophilic cation 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolcarbocyanine iodide (JC-1) revealed linear correlation between the median orange fluorescence (FL2) due to J-aggregate formations and MMP values measured by TPP+. No correlation was detected with the green fluorescence (FL1) emission. A significantly higher correlation appeared between the FL2/FL1 ratio and MMP values. Within the same mitochondrial population, cytofluorimetric analysis revealed the presence of various classes of organelles with different MMP, whose distribution was dependent on metabolic condition. The highest functional heterogeneity was found in deenergized mitochondria, while the highest homogeneity was observed during the first phase of the phosphorylative process. Thus, these data suggest that the cytofluorimetric use of JC-1 provides direct experimental evidence for the hypothesis of functional mitochondrial heterogeneity, at least with respect to their membrane potential.

Electromembrane systems in conditions of concentration polarization: new developments in the rotating membrane disk method

The advantages of the rotating membrane disk (RMD) method, which allows the examination of electromembrane sustem (EMS) polarization under conditions of uniform availability of both surfaces of the membrane, are shown. To obtain accurate information on the values of membrane potential (and thus on the solution concentration near the membrane surfaces), diffusional relaxation in combination with a RMD has been investigated. Reversal regimes are studied while using the RMD. To calibrate the cell with flowing solution, an apparatus with a RMD is used. To determine the selectivity value of ion-exchange membranes under limiting current density conditions, a method is developed whereby the rotating membrane allows a constant value of diffusion layers to be preserved. Different kinds of ion-exchange membrane (CMV, AMV, Neosepta, Flemion, Ionac, Rhône-Poulenc, MC-40, MA-40, MF-4SC) have been studied. It is found that, under limiting current density conditions, the selectivity value P in the majority of the membranes (except the hydrophobic perfluorinated membranes Flemion and MF-4SC) is considerably lower than it would be under equilibrium conditions. The effect on membrane selectivity produced by excess hydrostatic pressure on one side of the membrane disk is noted.

Changes in the plasmatic membrane characteristics during microsomal monooxygenase induction in the liver of adult and old rats

The experiments on adult (6–8 months) and old (24–26 months) male Wistar rats have shown that treatment of animals with phenobarbital results in a significant increase in hepatic microsomal enzyme content, plasmatic membrane Na +,K +-ATPase activities and the elevation of hepatocyte membrane potential value. It is presumed that the changes in plasmatic membrane characteristics during microsomal monooxygenase induction are related to the synthesis of specific intracellular factors (invertors). This assumption was verified by the experiments with ‘cellular hybrid’ system (cytosol — plasmatic membranes). Using this cross-systems, it was shown that the hepatocyte cytosol of rats treated with phenobarbital produced Na +,K +-ATPase activity. The extent of Na +,K +-ATPase activation was essentially lower when cytosol derived from old rat hepatocytes was used. The presence of specific factors that activated Na +,K +-ATPase in hepatocyte plasmatic membrane was also discovered in blood serum of induced adult and old rats.

Electrical properties of the cerebral prothoracicotropic hormone cells in diapausing and non-diapausing pupae of the tobacco hornworm, Manduca sexta.

Prothoracicotropic hormone (PTTH) is an insect brain neuropeptide that is a primary factor regulating an insect development. Curtailment of its release is thought to be responsible for the pupal diapause of tobacco hornworm, Manduca sexta. The cell synthesizing and secreting the PTTH has been identified as a pair of neurosecretory cells in the pars lateralis on each brain hemisphere. Using intracellular recording techniques, we have demonstrated electrical properties of the PTTH cells in different physiological status, i.e., diapausing and developing pupae. In diapausing pupae, they showed threshold value increasing and input resistance decreasing with the progress of diapausing state, indicating that they were getting unexcitable. Spontaneous action potentials and excitatory postsynaptic potentials (EPSPs) were rarely observed in deeply diapausing state. Non-diapausing PTTH cells were almost silent except day-2, showing rather constant values of electrical properties. On day-2, a significant proportion of the cells had spontaneous action potentials, showing less negative membrane potential values than inactive cells. Exclusively inhibitory postsynaptic potentials (IPSPs) were observed in significant numbers of the cells during the period from day-2 to day-5. On the basis of the results obtained, we proposed a working hypothesis that electrical activities of the PTTH cell may be primarily regulated by its membrane properties which are further modulated by the synaptic mediation.

Cell membrane stretch in osteoclasts triggers a self-reinforcing Ca2+ entry pathway.

Many cell types respond to mechanical membrane perturbation with intracellular Ca2+ responses. Stretch-activated (SA) ion channels may be involved in such responses. We studied the occurrence as well as the underlying mechanisms of cell membrane stretch-evoked responses in fetal chicken osteoclasts using separate and simultaneous patch-clamp and Ca2+ imaging measurements. In the present paper, evidence is presented showing that such responses involve a self-reinforcing mechanism including SA channel activity, Ca(2+)-activated K+ (KCa) channel activity, membrane potential changes and local and general intracellular Ca2+ ([Ca2+]i) increases. The model we propose is that during membrane stretch, both SA channels and KCa channels open at membrane potential values near the resting membrane potential. SA channel characterization showed that these SA channels are permeable to Ca2+. During membrane stretch, Ca2+ influx through SA channels and hyperpolarization due to KCa channel activity serve as positive feedback, leading ultimately to a Ca2+ wave and cell membrane hyperpolarization. This self-reinforcing mechanism is turned off upon SA channel closure after cessation of membrane stretch. We suggest that this Ca2+ entry mechanism plays a role in regulation of osteoclast activity.

Characterization of a transient outward current in a rapidly adapting insect mechanosensory neuron.

This paper describes the first voltage-clamp recordings from an arthropod cuticular sensory neuron. In the femoral tactile spine neuron of the cockroach Periplaneta americana, a rapidly activating and inactivating outward current, IA, appeared when the neuron was hyperpolarized for a short period before a depolarizing test pulse. IA could be separated from the other outward currents using 5 mM 4-aminopyridine (4-AP), which specifically blocked it. Tetraethylammonium (TEA), (50 mM) did not remove IA, but decreased the steady-state outward current by about 50%. The threshold for IA activation was about -75 mV. The minimum activation and inactivation time constants were approximately 0.2 ms and 15 ms, respectively. The voltage dependencies of activation and inactivation were well fit-ted by Boltzmann distributions, giving values of membrane potential at half-maximal activation (V50) equal to -56.5 mV and an equivalent gating charge of n = 3.9 for activation and V50 = -86.7 mV and n = 3.4 for inactivation. In current-clamp recordings, 4-AP reversibly reduced the cell's normal adaptation by lowering the threshold for action potentials, but did not affect the amplitude or duration of single action potentials. These results indicate that IA plays a role in short-term adaptation by opposing membrane depolarization and reducing the spike frequency during maintained stimulation.

Bovine serum albumin selectively increases the low-voltage-activated calcium current of NG108-15 neuroblastoma × glioma cells

The effect of bovine serum albumin (BSA) on the neuronal Ca 2+ current ( I Ca) has been studied in NG108-15 cells. BSA selectively increased the low-voltage-activated (l-v-a), transient I Ca but did not affect the high-voltage-activated (h-v-a) I Ca. The increase of l-v-a I Ca was partly due to a small shift of its activation curve to more negative values of membrane potential and partly to an increase of the maximal activatable current. Fatty-acid-free (FA-free) BSA was about equally effective as ordinary BSA; the smallest effective concentration of FA-free BSA was between 0.01 and 0.1 mg/ml.

The effect of respiration on the permeability of the mitochondrial membrane to ions

1. The rates of cation uptake, for either organic cations such as tetrapropylammonium, TPA +, at variable tetraphenylboron concentrations, TPB −, or inorganic cations such as Mn 2+, or K + plus valinomycin, have been measured in mitochondria either respiring, under uncoupler titrations, or non-respiring, under variable K + diffusion potentials. 2. The flow-force relationship for the respiration-coupled ion fluxes during titrations with uncouplers is almost identical to that obtained for the K +-diffusion driven fluxes. Similar results are obtained when TPA + is replaced with inorganic cations, either monovalent such as K + (+ valinomycin), or divalent such as Mn 2+. 3. By applying the Eyring analysis, as developed by Garlid et al. (Garlid, K.D., Beavis, A.D. and Ratkje, S.K. (1989) Biochim. Biophys. Acta 976, 109–121), from the flux-voltage relationships the values for the permeability coefficients and for the energy barriers have been obtained for the transport of the ion pair TPA +-TPB −, of Mn 2+ and of K + plus valinomycin, in non-respiring and in respiring, coupled and uncoupled, mitochondria. 4. The findings that the rates of respiration-coupled ion fluxes, at all values of membrane potential, are similar to the rates of the K + diffusion potential-coupled ion fluxes and the similar pattern of the flux-voltage relationships during the titrations with uncouplers and artificial gradients indicate that the membrane permeability for ions is not modified by respiration.

Interdependence of K+ and glutamate accumulation during osmotic adaptation of Escherichia coli.

Escherichia coli responds to an increase in medium osmolarity by accumulating K+ and glutamate. At low osmolarity a large fraction of cytoplasmic K+ serves to balance charge on macromolecular anions. That fraction of K+ is here referred to as "bound," as distinguished from "free" K+ that serves to balance charge of small anions. At higher osmolarity where cytoplasmic K+ increases markedly, the bound fraction decreases but the absolute amount of bound K+ expressed per unit of dry weight increases. The increase in bound K+ can be explained largely by the reduction of cytoplasmic putrescine at high osmolarity. At high osmolarity, glutamate is the major cytoplasmic anion, equal to at least 70% of free cytoplasmic K+. A sudden increase in the osmolarity of the medium stimulates glutamate synthesis with a lag of only about a minute; glutamate synthesis is almost totally dependent on K+ uptake. The high rate of flow of nitrogen through the glutamate pool under control conditions of growth at low osmolarity indicates that glutamate accumulation immediately after shift to high osmolarity must be due to inhibition of utilization of glutamate in the synthesis of other nitrogen-containing compounds rather than stimulation of glutamate synthesis. In agreement with this reasoning we find the kinetics of glutamate accumulation to be independent of the specific path of synthesis, whether by glutamate dehydrogenase or by glutamate synthase. Synthesis of glutamate appears to be required to attain normal values of the electrical membrane potential after shift to high osmolarity.

Regulation of a voltage-dependent, calcium-activated K conductance by cyclic GMP in dissociated flounder enterocytes.

Enterocytes from the winter flounder (Pseudopleuronectes americanus) were isolated by collagenase digestion and maintained in flounder Ringer's solution. Whole cell currents were studied using the amphotericin-perforated whole-cell patch clamp technique. The mean resting membrane potential and capacitance values or dissociated cells were -45 +/- 7 mV and 5 +/- 0.4 pF, respectively. Enterocytes held at -20 mV and treated with 1 mumol.l-1 ionomycin exhibited outward currents when cells were stepped through a series of voltages from -60 to +110 mV. The reversal potential of this current in flounder Ringer's solution was -55 mV and the voltage at which half-maximal activation occurred was +20 mV. Voltage-dependent inhibition of outward current was observed at +60 mV and above. When cells were bathed in symmetric K Ringer's solution the reversal potential shifted to zero mV and no inhibition of current was observed at voltages between -60 and 140 mV. When the holding potential of the cell was changed from -20 to -80 mV and stepped from -60 to +110 mV, a second [previously characterized, O'Grady et al. (1991)] K current with delayed-rectifier properties was identified. This observation demonstrated that the delayed rectifier K channel and the Ca(2+)-activated K channel described in this study exist in the same cell. Extracellular addition of 2 mmol.l-1 Ba2+ to cells bathed in symmetric K Ringer's solution resulted in nearly complete inhibition of outward current. Charybdotoxin produced only minor effects on this current.(ABSTRACT TRUNCATED AT 250 WORDS)