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T Mitsuiye

Publications and source records attributed to T Mitsuiye.

30 records · Page 2Linked to original sources
PubMed1990

Maximum open probability of single Na+ channels during depolarization in guinea-pig cardiac cells.

Single Na+ channel currents were recorded from guinea-pig ventricular cells in cell-attached patches. The ensemble average current (I) of multi-channel recordings was used to calculate the variance (sigma 2) of current fluctuations around the mean in individual current recordings. The relationship between sigma 2/I and I was linear and allowed estimation of the number of functional channels in the patch of membrane. The unitary amplitude of channel current obtained from the relation sigma 2/I-I was in agreement with that obtained directly by measuring the original records. The number of channels determined at different depolarizing pulses was almost constant in a given patch. The value was nearly equal to that of the maximum current, measured at high depolarizing potentials when most channels are open, divided by the unitary current. The open probability of the channels at the peak time of mean current was calculated based on the estimated number of channels. It increased with increasing depolarization and saturated at about 0.6 at test potentials above -20 mV. The inactivation time-course of the mean current was fitted by a sum of two exponentials. The current amplitude extrapolated to time zero was much larger than the current which could be generated by all channels. This indicates that the inactivation of the Na+ channel develops with delay after the onset of depolarization. The finding is in agreement with a model in which the inactivation rate is accelerated with activation of the Na+ channel.

Animals↗
PubMed1990

Negative shift of cardiac Na+ channel kinetics in cell-attached patch recordings.

Na+ channel kinetics were studied by recording single-channel currents in the cell-attached patch configuration of the patch-clamp technique in single ventricular cells isolated from guinea pig hearts. The inactivation time course of ensemble currents was accelerated, and the peak amplitude increased temporarily and then decreased within a few minutes after the gigaohm seal formation. After reaching a new steady state, the inactivation-voltage relation was found to have shifted to more negative potentials. The potential of half-maximal inactivation was more negative by 20-31 mV from the resting potential or between -96 and -112 mV. The voltage dependency of the channel activation also shifted. Although the cell membrane was depolarized using the whole cell patch-clamp electrode and single-channel currents were recorded with an independent cell-attached electrode, the shift of the inactivation curve was also evident. Complete removal of Ca2+ using 5 mM ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid in the pipette solution failed to prevent the shift. Increasing Ca2+ to 10 mM, however, reduced magnitude of the shift significantly. Involvement of an increased membrane fluidity and surface potential of the glass pipette to the shift is discussed.

Animals↗
PubMed1989

The Mg2+ block and intrinsic gating underlying inward rectification of the K+ current in guinea-pig cardiac myocytes.

1. The blockade by Mg2+ and intrinsic gating of the channel, which underlie the rectification of the inward rectifier K+ current, was investigated using the oil-gap voltage clamp method in isolated guinea-pig ventricular cells. 2. The inward rectifier K+ current was isolated by subtracting trans-gap currents recorded at an extracellular K+ concentration ([K+]o) of 0 mM from those obtained at 14 mM [K+]o in the presence of a given concentration of intracellular Mg2+ ([Mg2+]i). The reversal potential (V0) of the difference current was near the equilibrium potential for K+ (EK). 3. On repolarization across EK, the inward rectifier K+ current showed a rapid exponential increase. The time constant decreased with increasing hyperpolarization, but it was independent of both [Mg2+]i and the preceding depolarization. 4. When the pre-pulse potential was made progressively positive between V0-20 and V0 + 30 mV, the amplitude of the time-dependent component became larger and the preceding current jump decreased at any [Mg2+]i. With pre-pulses more positive than V0 + 40 mV, the time-dependent component started from almost the zero current level at 2 microM [Mg2+]i. At higher [Mg2+]i (350, 500 and 3000 microM), however, the time-dependent component became smaller as the pre-pulse potential was made more positive than V0 + 40 mV. 5. When the membrane was depolarized from a potential of full activation at 2 microM [Mg2+]i, the initial jump in the outward current was ohmic and was followed by an exponential decay. The time-dependent component of the inward current, recorded on repolarization after increasing durations of the preceding depolarization, developed as the outward current decayed. The time constants of both processes were in good agreement. 6. At 500 microM [Mg2+]i, the outward current on depolarization was instantaneously rectified. The time-dependent component recorded on repolarization developed with prolongation of the pre-pulse with a time course slower than at 2 microM [Mg2+]i. The envelope time course became slower as the potential of the depolarization became more positive. 7. Lowering the temperature from 23 to 15 degrees C slowed the time-dependent current with an apparent Q10 of about 3.5 at V0. 8. Based on the experimental data, kinetic parameters were estimated for a model of Mg2+ block, which well simulated the inward-going rectification of the K+ current. 9. It is concluded that the instantaneous inward rectification on depolarization is due to the Mg2+ block at physiological [Mg2+]i.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1987

A new oil-gap method for internal perfusion and voltage clamp of single cardiac cells.

(1.) We designed a new technique to achieve fast voltage clamp, combined with internal perfusion. The single guinea-pig cardiac cell, dissociated by collagenase treatment, was stretched across an oil-gap (30-40 micron wide) from a pool of Tyrode solution to a pool of internal solution. Part of the cell membrane was disrupted in the internal solution by crushing on the cell, a tapered tip of a glass capillary. Through the open end, the intracellular medium was equilibrated with test solutions and electrical current was injected for the voltage clamp of the membrane in the Tyrode pool. (2.) The capacitive transient on stepping the membrane potential decayed with a time constant of 10-60 microseconds, depending on the capacitive area (20-80 pF). The time course was a single exponential in 46% of the atrial cells and in 66% of the ventricular cells. In these tissues the series resistance, approximated by a ratio of the time constant and Cm, was 686 +/- 180 k omega (n = 37) in the ventricular cells or 812 +/- 143 k omega (n = 18) in the atrial cells. The stable seal resistance (Rseal) established in the oil-gap was around 33 M omega in the ventricular cells and 100 M omega in the atrial cells. (3.) A rapid increase in the inward current followed by a slow decay was observed on repolarization over the range negative to the potassium equilibrium potential. From the inward rectification of both peak and late currents and suppressive effects of Cs+ on the current, the current changes were attributed to activation and inactivation of the inward rectifier K channel.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1986

Reduction of the voltage-dependent calcium current in Aplysia neurons by pentobarbital.

Effects of pentobarbital on the calcium current of Aplysia neurons were investigated under current- and voltage-clamp conditions using the conventional two-microelectrode technique. Pentobarbital attenuated the progressive broadening of repeated action potentials of somata, suggesting a reduction in the calcium current. When calcium ion was replaced with barium ion in the perfusing solution, in which neither sodium nor potassium ions carried transmembrane currents, the barium current (IBa) which flowed through the calcium channel of the cell membrane was generated by depolarizing pulses of several hundred milliseconds applied every 1 min from a holding potential of -50 mV. The IBa was not affected by tetrodotoxin (30 microM). The current was decreased by pentobarbital (0.1-5 mM) in a dose-dependent manner. The inhibition was much greater at a lower pH of the perfusate, indicating that the uncharged form of the agent was responsible. The voltage-dependent inactivation of the IBa proceeded with two time constants [190 +/- 21 and 2020 +/- 146 msec (N = 4) at -10 mV], both of which were shortened by adding 1 mM pentobarbital [to 120 +/- 18 and 540 +/- 51 msec (N = 4), respectively]. The IBa recovered from the inactivation with two time constants [60 +/- 7 and 871 +/- 76 msec (N = 3) at -50 mV]. The anesthetic (1 mM) prolonged both of them, to 124 +/- 20 and 1480 +/- 172 msec (N = 3), respectively, resulting in a use-dependent depression of the current at 2-Hz stimulation. Pentobarbital reduced the IBa to a greater extent when the holding potential was more positive (-30 instead of -50 mV), indicating a higher affinity of the drug to the inactivated state of the channel. These findings suggest that the attenuation of the progressive broadening of successive spikes by pentobarbital is due to a decrease in the voltage- and time-dependent calcium current, ending in depression of transmitter release from the nerve terminal.

Action Potentials↗
PubMed1985

Potassium-related membrane currents in the bullfrog atrial muscle differentiated in the presence of barium.

Effects of Ba on the potassium-related currents were studied on the bullfrog atrial muscle under voltage clamp with double sucrose-gap techniques. Ba, in a dose over 0.1 mM, abolished the anomalous rectification of the membrane by inhibiting the background current which reversed sign nearly at the K equilibrium potential (IK1). Ba, thus reducing the K-depletion current for hyperpolarizations, revealed the presence of funny inward current (If or Ih) in the proper atrial muscle. An increase in [K]0 increased If, and the current showed a threshold at about -80 mV and was saturated at above -160 mV in 5 mM [K]0. The delayed outward current (Ix) for depolarizations was also depressed by Ba. The depression occurred in a voltage- and time-dependent manner, manifesting an unblocking for stronger depolarizations. An analysis of the current tail, however, disclosed that low concentrations of Ba (up to 0.1 mM) inhibited the accumulation component (Ia) of the current without diminishing the next slow component of Ix (Ixs). The remaining Ixs showed a reversal potential of -82 mV, suggesting that this current is largely carried by potassium ions. These data clearly show that in the presence of Ba, If and Ixs can be differentiated from other membrane currents in the frog atrial muscle.

Animals↗
PubMed1984

Adrenergic-cholinergic interactions on membrane potential of K+ -depolarized ventricular muscle.

In guinea pig ventricular muscles exposed to K+-rich (27 mM) Tyrode solution containing 0.2 mM Ba, catecholamine elicited a slight depolarization of the resting membrane. Application of acetylcholine (ACh) during this catecholamine-induced response caused a repolarization, and removal of ACh induced a transient enhancement in the depolarization (rebound). These effects of ACh were abolished by atropine. Application of ACh alone and its removal had little effect on the membrane potential. Like the catecholamine-induced depolarization, the rebound depolarization after ACh removal was inhibited by slow channel blockers. Thus the rebound was attributed at least in part to enhanced changes in the catecholamine-sensitive conductance, i.e., a beta-receptor-mediated increase in the slow channel conductance. In driven muscles perfused with normal Tyrode solution, there was a rebound increase in twitch tension when ACh was removed in the presence of catecholamine, and this rebound was accompanied by an "extra" elevation of the action potential plateau. Thus cessation of the stimulation of myocardial muscarinic receptors may transiently lead to an enhanced activity of the beta-adrenoceptor-slow channel system in guinea pig ventricular muscle.

Acetylcholine↗
PubMed1984

Transient increase in the slow inward current following acetylcholine removal in catecholamine-treated guinea-pig Purkinje fibers.

In voltage-clamped guinea-pig Purkinje fibers, removal of acetylcholine (ACh) in the presence of isoproterenol produced a rebound increase in the slow inward current. This effect of ACh was abolished by atropine. These results are consistent with the hypothesis that cessation of the stimulation of myocardial muscarinic receptors transiently leads to enhanced activity of the beta-receptor/slow channel system in this tissue.

Acetylcholine↗
PubMed1983

Depolarization produced by catecholamines in guinea-pig ventricular muscle cells exposed to potassium-rich media and its dependence on temperature.

Catecholamines (isoproterenol, adrenaline, and noradrenaline) elicited a small decrease in the resting potential of guinea-pig ventricular muscle cells depolarized by 27 mM K. This catecholamine-induced depolarization (CAD) was enhanced and often led to an automatic activity, when the membrane shunting conductance was reduced by application of 0.05 to 0.2 mM Ba. CAD was blocked by Mn (1 to 2 mM), verapamil (0.5 to 1 X 10(-5) M), and propranolol (0.1 to 1 X 10(-5) M), but not by phentolamine (10(-5) M). CAD did not develop when both Ca and Ba were absent in the bathing solution, but persisted when Sr was present. These results are consistent with the hypothesis that CAD was due at least partly to an increase in the slow channel conductance that was initiated by catecholamine/beta-receptor interaction. CAD was markedly enhanced at low temperatures (21 to 25 degrees C), and such was characterized by slow repolarization after drug withdrawal. Propranolol, when applied after catecholamine, exerted no appreciable effect on this slow repolarization. This beta-blocker abolished CAD at low temperature, if applied prior to catecholamine. Methylxanthines (2 to 5 mM caffeine or theophylline) produced a depolarization similar to that seen with CAD, and the rate of repolarization after drug withdrawal also slowed at low temperature. The slow repolarization of CAD at low temperature appeared to reflect a slowing in the postreceptor metabolic processes responsible for deactivation of the slow channel that was sensitive to beta-receptor stimulation.

Animals↗