The toxic effects of ouabain: A voltage-clamp study

1982 ◽  
Vol 60 (9) ◽  
pp. 1153-1159 ◽  
Author(s):  
Y. Deslauriers ◽  
E. Ruiz-Ceretti ◽  
O. F. Schanne ◽  
M. D. Payet
Keyword(s):  
Action Potential ◽  
Voltage Clamp ◽  
Sodium Current ◽  
Negative Inotropic Effect ◽  
Slow Inward Current ◽  
Inward Current ◽  
Toxic Concentration ◽  
Voltage Curve ◽  
Current Voltage Curve ◽  
High Concentrations

The electrophysiologic effects of a toxic concentration of ouabain (10−5 M) were studied in frog atrial trabeculae. The toxic concentration was determined by the appearance of a negative inotropic effect and an increase in basal tension. Current- and voltage-clamp measurements were performed. Ouabain did not alter the passive electrical properties of the preparation. Under current-clamp conditions the membrane depolarized and the action potential amplitude as well as its maximum rate of rise decreased. The current–voltage curve for the fast inward current was shifted toward more positive potentials and the maximum sodium current decreased. The maximum sodium conductance was also reduced. The process of reactivation of the fast inward current was accelerated. The slow inward current and the maximum slow conductance also decreased under ouabain. These effects could explain the negative inotropic action of high concentrations of glycosides, as well as the action potential changes observed by several investigators. They also help to understand the arrhythmogenic effects of high concentrations of digitalis.

scholarly journals Two Inward Currents in Frog Atrial Muscle

1971 ◽  
Vol 58 (5) ◽  
pp. 523-543 ◽  
Author(s):  
Merrill Tarr
Keyword(s):  
Action Potential ◽  
Voltage Clamp ◽  
Ionic Currents ◽  
Ringer Solution ◽  
Slow Inward Current ◽  
Slow Phase ◽  
Free Solution ◽  
Rapid Phase ◽  
Inward Current ◽  
Atrial Tissue

The double sucrose-gap voltage-clamp technique was applied to frog atrial tissue to investigate the ionic currents responsible for the action potential in this tissue. Membrane depolarization elicited two distinct components of inward current when the test node was exposed to normal Ringer solution: a fast inward current and a slow inward current. The fast inward current appeared to be carried by sodium ions, since it was rapidly abolished by exposure of the fiber to Na+-free solution or tetrodotoxin but persisted on exposure to Ca++-free solution. In contrast, in the majority of the preparations the slow inward current appeared to be primarily carried by calcium ions, since it was abolished on exposure of the fiber to Ca++-free solution but persisted on exposure to Na+-free solution. Action potential data supported the voltage-clamp findings. The normal action potential shows two distinct components in the upstroke phase: an initial rapid phase of depolarization followed by a slower phase of depolarization reaching the peak of the action potential. Abolition of the fast inward current resulted in abolition of the initial rapid phase of depolarization. Abolition of the slow inward current resulted in abolition of the slow phase of depolarization. These data support the hypothesis that two distinct and different ionic mechanisms contribute to the upstroke phase of the action potential in frog atrial tissue.

scholarly journals Contribution of L-type Ca2+current to electrical activity in sinoatrial nodal myocytes of rabbits

1999 ◽  
Vol 276 (3) ◽  
pp. H1064-H1077 ◽  
Author(s):  
E. Etienne Verheijck ◽  
Antoni C. G. van Ginneken ◽  
Ronald Wilders ◽  
Lennart N. Bouman
Keyword(s):  
Action Potential ◽  
Calcium Current ◽  
Sodium Current ◽  
Delayed Rectifier ◽  
Current Clamp ◽  
Patch Clamp Technique ◽  
Inward Current ◽  
Delayed Rectifier Current ◽  
Diastolic Depolarization ◽  
Recovery From Inactivation

The role of L-type calcium current ( I Ca,L) in impulse generation was studied in single sinoatrial nodal myocytes of the rabbit, with the use of the amphotericin-perforated patch-clamp technique. Nifedipine, at a concentration of 5 μM, was used to block I Ca,L. At this concentration, nifedipine selectively blocked I Ca,L for 81% without affecting the T-type calcium current ( I Ca,T), the fast sodium current, the delayed rectifier current ( I K), and the hyperpolarization-activated inward current. Furthermore, we did not observe the sustained inward current. The selective action of nifedipine on I Ca,L enabled us to determine the activation threshold of I Ca,L, which was around −60 mV. As nifedipine (5 μM) abolished spontaneous activity, we used a combined voltage- and current-clamp protocol to study the effects of I Ca,L blockade on repolarization and diastolic depolarization. This protocol mimics the action potential such that the repolarization and subsequent diastolic depolarization are studied in current-clamp conditions. Nifedipine significantly decreased action potential duration at 50% repolarization and reduced diastolic depolarization rate over the entire diastole. Evidence was found that recovery from inactivation of I Ca,L occurs during repolarization, which makes I Ca,L available already early in diastole. We conclude that I Ca,L contributes significantly to the net inward current during diastole and can modulate the entire diastolic depolarization.

A voltage-clamp analysis of currents underlying cyclic AMP-induced membrane modulation in isolated peptidergic neurons of Aplysia

1984 ◽  
Vol 52 (2) ◽  
pp. 340-349 ◽  
Author(s):  
L. K. Kaczmarek ◽  
F. Strumwasser
Keyword(s):  
Cell Culture ◽  
Cyclic Amp ◽  
Voltage Clamp ◽  
Cultured Cells ◽  
Sodium Current ◽  
Negative Slope ◽  
Spontaneous Discharge ◽  
Inward Current ◽  
Outward Currents ◽  
Bag Cell Neurons

A variety of chemical and electrophysiological evidence indicates that the onset of afterdischarge and the subsequent profound enhancement of spike broadening that occur in the bag cell neurons of Aplysia are related to an increase in adenosine 3',5'-monophosphate-(cAMP) dependent protein phosphorylation. We have now used a two-electrode voltage clamp to study the properties of isolated bag cell neurons in cell culture and their response to 8 benzylthio-cAMP (8BTcAMP) and N6-n-butyl 8BTcAMP. These membrane-permeant and phosphodiesterase-resistant cAMP analogs induce spontaneous discharge and spike broadening in both the intact bag cell cluster and isolated bag cell neurons in cell culture. The dominant inward current in these cultured cells was found to be the calcium current, Ica, which was abolished by Co2+ (20 mM) or Ni2+ (10 mM) and could be observed in Na+-free media. In a minority of cells (2 of 12), in normal ionic media, a transient inward current was observed that was unaffected by Co2+ and Ni2+ and probably represents a sodium current. The three characterized potassium currents, the delayed rectifying current IK, the calcium-dependent current IC, and the early transient current IA, distinguished by their differing pharmacological and voltage-activation properties, were present in all healthy cells. Three effects of the cyclic AMP analogs (0.5 mM) on the electrical properties of these cells were 1) the emergence of a region of negative slope resistance in the steady-state I-V relations, 2) a depression of the net sustained outward currents due to depolarizing commands, and 3) a marked reduction in IA. When outward currents had been largely suppressed using high concentrations of tetraethylammonium (TEA) ions (100-460 mM) no effects of the cyclic AMP analogs could be observed on peak inward currents using NA+ and Ca2+ or Ba2+ as carriers of inward current. At least part of these electrical effects of the cyclic AMP analogs could be accounted for by a depression of a delayed potassium current and the A current.

Effects of ischemic conditions and reperfusion on depolarization-induced automaticity

1988 ◽  
Vol 255 (5) ◽  
pp. H992-H999 ◽  
Author(s):  
R. Mohabir ◽  
G. R. Ferrier
Keyword(s):  
Membrane Potential ◽  
Action Potential ◽  
Action Potentials ◽  
Cycle Length ◽  
Slow Inward Current ◽  
Potential Range ◽  
Ischemia And Reperfusion ◽  
Slow Response ◽  
Inward Current ◽  
High Membrane Potential

The inducibility of slow-response automaticity was assessed during ischemic conditions and reperfusion by application of extracellular current. Isolated canine Purkinje fibers were depolarized to membrane potentials less than -65 mV to elicit depolarization-induced automaticity (DIA). Ischemic conditions increased the cycle length of DIA and, in some tissues, prevented sustained DIA or completely abolished DIA. The magnitude of depolarization required to elicit DIA also increased. Inhibition of DIA occurred at a time when action potential plateaus were abbreviated. The effect of reperfusion on DIA was biphasic. Initial reappearance of DIA was followed by inhibition and reduction of the membrane potential range over which DIA could be elicited. Plateaus of action potentials initiated at high membrane potential were abbreviated at this time. DIA returned again as reperfusion effects dissipated. Phasic changes in the inducibility of DIA may represent changes in availability of the slow inward current and may regulate the timing and types of arrhythmic activity occurring with ischemia and reperfusion.

Fast Na+ channels in smooth muscle from pregnant rat uterus

1992 ◽  
Vol 70 (4) ◽  
pp. 491-500 ◽  
Author(s):  
Nicholas Sperelakis ◽  
Yoshihito Inoue ◽  
Yusuke Ohya
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Current Density ◽  
Sodium Current ◽  
Muscle Cells ◽  
Slow Inward Current ◽  
Rat Uterus ◽  
Pregnant Rat ◽  
Inward Current ◽  
Channel Current

Smooth muscle cells normally do not possess fast Na+ channels, but inward current is carried through two types of Ca2+ channels: slow (L type) Ca2+ channels and fast (T type) Ca2+ channels. Whole-cell voltage clamp was done on single smooth muscle cells isolated from the longitudinal layer of the 18-day pregnant rat uterus. Depolarizing pulses, applied from a holding potential of −90 mV, evoked two types of inward current, fast and slow. The fast inward current decayed within 30 ms, depended on [Na]o, and was inhibited by tetrodotoxin (TTX) (K0.5 = 27 nM). The slow inward current decayed slowly, was dependent on [Ca]o (or Ba2+), and was inhibited by nifedipine. These results suggest that the fast inward current is a fast Na+ channel current and that the slow inward current is a Ca2+ slow channel current. A fast-inactivating Ca2+ channel current was not evident. We conclude that the ion channels that generate inward currents in pregnant rat uterine cells are TTX-sensitive fast Na+ channels and dihydropyridine-sensitive slow Ca2+ channels. The number of fast Na+ channels increased during gestation. The averaged current density increased from 0 on day 5, to 0.19 on day 9, to 0.56 on day 14, to 0.90 on day 18, and to 0.86 pA/pF on day 21. This almost linear increase occurs because of an increase in the fraction of cells that possess fast Na+ channels. The Ca2+ channel current density was also higher during the latter half of gestation. These results indicate that the fast Na+ channels and Ca2+ slow channels in myometrium become more numerous as term approaches, and we suggest that the fast Na+ current may be involved in spread of excitation. Isoproterenol (β-agonist) did not affect either ICa(s) or INa(f), whereas Mg2+ (K0.5 = 12 mM) and nifedipine (K0.5 = 3.3 nM) depressed ICa(s). Oxytocin had no effect on INa(f) and actually depressed ICa(s) to a small extent. Therefore, the tocolytic action of β-agonists cannot be explained by an inhibition of ICa(s), whereas that of Mg2+ can be so explained. The stimulating action of oxytocin on uterine contractions cannot be explained by a stimulation of ICa(s).Key words: sodium current, fast sodium current, calcium currents, myometrial smooth muscle cells, pregnant uterine muscle.

Slow inward current may produce many results attributed to IX1 in cardiac Purkinje fibers

1985 ◽  
Vol 249 (1) ◽  
pp. H122-H132
Author(s):  
J. M. Jaeger ◽  
W. R. Gibbons
Keyword(s):  
Action Potential ◽  
Outward Current ◽  
Purkinje Fiber ◽  
Slow Inward Current ◽  
Channel Blockers ◽  
Inward Current ◽  
Purkinje Fibers ◽  
Cardiac Purkinje Fibers ◽  
Current Voltage ◽  
Current Voltage Relation

We have tried to answer two fundamental questions concerning the outward current IX1 of cardiac Purkinje fibers. 1) Is it possible that current changes identified as arising from IX1 in voltage-clamp experiments are actually manifestations of changes in the slow inward current (Isi); and 2) is IX1 in fact required to produce the electrical phenomena attributed to it? Isi behavior and the role of IX1 were explored using computer simulation. The Isi model produced current changes during depolarizations and hyperpolarizations from depolarized resting potentials like those attributed to IX1. It also produced a component of "tail currents" that behaved like IX1. If these current changes were analyzed, assuming that an outward current is responsible, the resulting kinetics and current voltage relation would be very similar to the kinetics and current voltage relation reported for IX1. Using the McAllister, Noble, and Tsien formulation of the Purkinje fiber action potential, we found that IX1 is not essential for repolarization of the reconstructed action potential nor is it needed to reproduce interval duration effects and the effects of applied current in that model. Data suggesting that calcium channel blockers reduce IX1 and that catecholamines increase IX1 may be explained as arising from changes in Isi. Thus many manifestations of IX1 can be explained as arising from unanticipated behavior of Isi, and IX1 does not necessarily play a key role in generating Purkinje fiber electrical activity.

Tl+-Ions: Comparison of the Effects on the Slow Inward Current and Contractility of Ventricular Tissues

1982 ◽  
Vol 37 (10) ◽  
pp. 1015-1022 ◽  
Author(s):  
J. Wiemer ◽  
R. Ziskoven ◽  
C. Achenbach
Keyword(s):  
Voltage Clamp ◽  
Current System ◽  
Slow Inward Current ◽  
Initial Increase ◽  
Purkinje Fibres ◽  
Cardiac Tissues ◽  
Inward Current ◽  
Current Voltage ◽  
Apparent Relationship ◽  
Time Courses

To conclude our investigation of thallium effects on cardiac tissues, we studied the slow inward current of sheep cardiac Purkinje fibres exposed to 10-7 to 10-5 ᴍ Tl+ for extended periods of up to 80 min. Our previous results had suggested a possible involvement of the slow inward current during thallium intoxication: a) the modification of contractility staircases observed during thallium exposure, b) action potential recordings of ventricular muscle, c) changes in spontaneous beating in sino-atrial preparations. The thallium levels chosen were between those yielding strong positive inotropic transients and those producing a marked long­term decay of contraction force.The slow inward current was measured using a conventional two-microelectrode-technique and the standard voltage clamp protocol for this current system. The experimental work was restricted to the determination of d∞, the kinetics of activation of the slow inward current and of īsi, the current voltage relation of the current system. This was necessary since the effects of thallium were known to be short-lived and therefore frequent repeat runs of the voltage clamp program had to be performed in order to obtain the time courses of possible transient changes.The results showed that the slow inward current was first increased and then declined at the low concentration of 10-7 ᴍ Tl+. At 10-5 m Tl+ the initial increase was smaller, whereas the decay of the slow inward current proceeded to lower values. Comparison with contractility measure­ments at the same concentrations of thallium showed a distinct parallelism between changes of the slow inward current and myocardial contractility. Despite this apparent relationship, we do not conclude that the contractile events are primarily a result of changes of the slow inward current, since thallium does not seem to specifically alter the parameters of the slow inward current at the membrane level.

Voltage Clamp Studies on the Slow Inward Current during the Excitation ofNitellopsis obtusa

1980 ◽  
Vol 31 (2) ◽  
pp. 589-595 ◽  
Author(s):  
M. GYENES ◽  
A. A. BULYCHEV ◽  
G. A. KURELLA
Keyword(s):  
Voltage Clamp ◽  
Slow Inward Current ◽  
Inward Current

scholarly journals Anomalous Rectification in the Squid Giant Axon Injected with Tetraethylammonium Chloride

1965 ◽  
Vol 48 (5) ◽  
pp. 859-872 ◽  
Author(s):  
Clay M. Armstrong ◽  
Leonard Binstock
Keyword(s):  
Action Potential ◽  
Outward Current ◽  
Giant Axon ◽  
Potassium Ion ◽  
Ion Concentration ◽  
High Potassium ◽  
Tetraethylammonium Chloride ◽  
Voltage Curve ◽  
Instantaneous Current ◽  
Current Voltage Curve

The injection of tetraethylammonium chloride into the giant axon of the squid prolongs the action potential and eliminates most of the late current under voltage-clamp. Experiments on fibers in an external medium of high potassium ion concentration demonstrate that injected tetraethylammonium chloride causes rectification of the instantaneous current-voltage curve for potassium by excluding outward current. This interference with the flow of outward potassium ion current underlies the prolongation of the action potential seen in tetraethylammonium-injected fibers.

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