Identification of and developmental changes in transient outward current in embryonic chick cardiomyocytes

1995 ◽  
Vol 7 (5) ◽  
pp. 1369 ◽  
Author(s):  
H Satoh
Keyword(s):  
Slow Component ◽  
Outward Current ◽  
Cell Voltage ◽  
Developmental Changes ◽  
Transient Outward Current ◽  
Embryonic Chick ◽  
Time To Peak ◽  
Ventricular Cells ◽  
Developmental Age ◽  
Voltage Dependent

Identification and developmental changes in the transient outward current (I(to)) in isolated embryonic chick ventricular cells (3, 10 and 17 days old) were examined using a whole-cell voltage clamp technique. Experiments were performed at room temperature (22 degrees C). Test pulses were applied between -40 and +50 mV from a holding potential of -60 mV. The I(to) was present (but small) and increased during development; the current density of I(to) at +40 mV was 3.5 +/- 0.5 pA/pF (n = 7) in 3-day cells, 4.2 +/- 0.9 pA/pF (n = 5) in 10-day cells, and 17.1 +/- 1.6 pA/pF (n = 5) in 17-day-old cells. The average capacitances also changed with developmental age; 12.0 +/- 2.0 pF (n = 8) in 3-day cells, 10.8 +/- 2.2 pF (n = 7) in 10-day cells, and 8.6 +/- 2.3 pF (n = 7) in 17-day cells. The I(to) was not always observed in all the prepared cells, and the number of cells possessing I(to) increased during development. The threshold potential was -30 mV in 17-day cells, and appeared to be displaced to more negative potential with developmental age. The time to peak decreased during development: 10.6 +/- 1.1 ms (n = 4) in 3-day cells, 6.7 +/- 0.5 ms (n = 5) in 10-day cells, and 5.4 +/- 0.6 ms (n = 5) in 17-day cells. The time decay of the inactivation phase for the I(to) had two exponentials; the fast component was increased by about 3-fold in 17-day cells, and the slow component was decreased by about 14% in both 10- and 17-day cells, as compared to 3-day cells. Addition of 3 mM 4-aminopyridine (4-AP) inhibited I(to) at +50 mV by 81.9 +/- 2.3% (n = 4, P < 0.001). These results indicate that the I(to), voltage-dependent and 4-AP-sensitive, exists even in young embryonic cardiomyocytes (but not in all cells), and increases during development, resulting in modulation of the action potential configuration.

scholarly journals Idiosyncratic Gating of HERG-like K+ Channels in Microglia

1998 ◽  
Vol 111 (6) ◽  
pp. 795-805 ◽  
Author(s):  
Peter S. Pennefather ◽  
Wei Zhou ◽  
Thomas E. DeCoursey
Keyword(s):  
Slow Component ◽  
Outward Current ◽  
Data Availability ◽  
Transient Outward Current ◽  
Maximal Conductance ◽  
Voltage Dependent ◽  
Simple Kinetic Model ◽  
Window Current ◽  
Herg Channels ◽  
Deactivation Process

A simple kinetic model is presented to explain the gating of a HERG-like voltage-gated K+ conductance described in the accompanying paper (Zhou, W., F.S. Cayabyab, P.S. Pennefather, L.C. Schlichter, and T.E. DeCoursey. 1998. J. Gen. Physiol. 111:781–794). The model proposes two kinetically distinct closing pathways, a rapid one favored by depolarization (deactivation) and a slow one favored by hyperpolarization (inactivation). The overlap of these two processes leads to a window current between −50 and +20 mV with a peak at −36 mV of ∼12% maximal conductance. The near absence of depolarization-activated outward current in microglia, compared with HERG channels expressed in oocytes or cardiac myocytes, can be explained if activation is shifted negatively in microglia. As seen with experimental data, availability predicted by the model was more steeply voltage dependent, and the midpoint more positive when determined by making the holding potential progressively more positive at intervals of 20 s (starting at −120 mV), rather than progressively more negative (starting at 40 mV). In the model, this hysteresis was generated by postulating slow and ultra-slow components of inactivation. The ultra-slow component takes minutes to equilibrate at −40 mV but is steeply voltage dependent, leading to protocol-dependent modulation of the HERG-like current. The data suggest that “deactivation” and “inactivation” are coupled through the open state. This is particularly evident in isotonic Cs+, where a delayed and transient outward current develops on depolarization with a decay time constant more voltage dependent and slower than the deactivation process observed at the same potential after a brief hyperpolarization.

Abnormalities of K+ and Ca2+ currents in ventricular myocytes from rats with chronic diabetes

1995 ◽  
Vol 269 (4) ◽  
pp. H1288-H1296 ◽  
Author(s):  
D. W. Wang ◽  
T. Kiyosue ◽  
S. Shigematsu ◽  
M. Arita
Keyword(s):  
Steady State ◽  
Action Potential ◽  
Ventricular Myocytes ◽  
Slow Component ◽  
Outward Current ◽  
Cell Voltage ◽  
Transient Outward Current ◽  
Inactivation Curve ◽  
Ionic Mechanisms ◽  
In Cells

Ionic mechanisms related to the prolongation of cardiac action potential in rats with chronic diabetes mellitus were studied using whole cell voltage-clamp techniques. Diabetes was induced by injection of streptozotocin (STZ; 65 mg/kg body wt) into the tail vein, and ventricular myocytes were isolated from STZ-injected rats (24-30 wk) and from age-matched normal rats. The current densities of transient outward current (Ito), a steady-state outward current, and L-type Ca2+ current (ICa) were significantly smaller in cells from diabetic animals. In addition, the kinetics of Ito of diabetic cells were modified. 1) The decay of Ito was well fitted by a sum of two exponential components in normal cells; there was only one (slow) component in the diabetic cells. 2) The steady-state inactivation curve of Ito in diabetic cells shifted by 5 mV in the negative direction. 3) Recovery from inactivation of Ito was slower in cells from diabetic animals. These alterations in Ito and the steady-state outward current can account for most of the action potential prolongation heretofore documented. The decrease of ICa may possibly be related to the depressed contraction seen in chronic diabetic mellitus.

Selective expression of transient outward currents in different types of acutely isolated hippocampal interneurons

1996 ◽  
Vol 76 (5) ◽  
pp. 3563-3567 ◽  
Author(s):  
S. H. Fan ◽  
R. K. Wong
Keyword(s):  
Outward Current ◽  
Pyramidal Cells ◽  
Cell Voltage ◽  
Transient Outward Current ◽  
Isolated Cells ◽  
Tissue Slices ◽  
Ca1 Region ◽  
Hippocampal Ca1 Region ◽  
Outward Currents ◽  
Voltage Dependent

1. Voltage-dependent outward currents in CA1 interneurons were studied with the use of whole cell voltage-clamp techniques. Tissue slices containing strata lacunosum-moleculare and radiatum (L-M-R regions) of the hippocampal CA1 region were prepared. Neurons were then isolated from these tissue slices with the use of an acute dissociation procedure. The morphologies of the isolated neurons were distinct from those of pyramidal cells and correlated with those of interneurons identified in the L-M-R regions after immunohistochemical stainings. 2. Total outward currents were elicited from the isolated cells by depolarization steps applied after a 300-ms hyperpolarization prepulse to 100 mV from a holding potential of 50 mV. Delayed outward currents were obtained by intercalating a 120-ms step at 55 mV between the hyperpolarizing prepulse and the depolarization. The intercalating step served to inactivate transient outward currents. Transient outward current were isolated by subtracting the delayed outward currents from the total outward currents. 3. Interneurons were subgrouped on the basis of their ability to produce transient outward current in response to the above protocol. 4. The two groups of interneurons possessed distinct morphological features. Cells producing transient outward currents had polygonal-shaped somata with thick primary processes that gave rise to smaller secondary processes at a short distance from the soma. Interneurons without activatable transient outward currents had somata that were not polygonal and they had more slender primary dendritic processes. 5. These results suggest that interneurons in the L-M-R regions can be divided into two groups on the basis of the presence or absence of voltage-dependent transient outward currents. The two groups of cells differentiated on this basis also have distinguishable morphological traits. The difference in the properties of the outward current may be a factor contributing to the variation in the firing pattern of recorded interneurons reported in previous studies.

Developmental change in fast Na channel properties in embryonic chick ventricular heart cells

1995 ◽  
Vol 73 (10) ◽  
pp. 1475-1484 ◽  
Author(s):  
Hideaki Sada ◽  
Takashi Ban ◽  
Takeshi Fujita ◽  
Yoshio Ebina ◽  
Nicholas Sperelakis
Keyword(s):  
Steady State ◽  
Voltage Clamp ◽  
Time Constant ◽  
Voltage Dependence ◽  
Cell Voltage ◽  
Developmental Changes ◽  
Heart Cells ◽  
Embryonic Chick ◽  
Whole Cell ◽  
Whole Cell Voltage Clamp

To assess developmental changes in kinetic properties of the cardiac sodium current, whole-cell voltage-clamp experiments were conducted using 3-, 10-, and 17-day-old embryonic chick ventricular heart cells. Experimental data were quantified according to the Hodgkin–Huxley model. While the Na current density, as examined by the maximal conductance, drastically increased (six- to seven-fold) with development, other current–voltage parameters remained unchanged. Whereas the activation time constant and the steady-state activation characteristics were comparable among the three age groups, the voltage dependence of the inactivation time constant and the steady-state inactivation underwent a shift in the voltage dependence toward negative potentials during embryonic development. Consequently, the steady-state (window current) conductance, which was sufficient to induce automatic activity in the young embryos, was progressively reduced with age.Key words: cardiac electrophysiology, whole-cell voltage-clamp experiments, fast Na currents, heart, development, developmental changes.

Ethacizin blockade of calcium channels: a test of the guarded receptor hypothesis

1989 ◽  
Vol 257 (5) ◽  
pp. H1693-H1704
Author(s):  
C. F. Starmer ◽  
A. I. Undrovinas ◽  
F. Scamps ◽  
G. Vassort ◽  
V. V. Nesterenko ◽  
...  
Keyword(s):  
Calcium Channels ◽  
Cell Voltage ◽  
Membrane Potentials ◽  
Specific Rate ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Channel Inactivation ◽  
Ventricular Cells ◽  
Channel Blocking ◽  
Voltage Dependent

The effect on calcium channels of the sodium channel antagonist, ethacizin, was studied in isolated frog ventricular cells using the whole cell voltage-clamp methodology. Ethacizin was found to block inward calcium current in a frequency-, voltage-, and concentration-dependent manner. The frequency-dependent blocking properties were modeled by considering the drug interaction with a voltage-dependent mixture of calcium channels harboring either an accessible or an inaccessible binding site. With repetitive stimulation, the pulse-to-pulse reduction in peak current is shown to be exponential, with a rate linearly related to the interstimulus interval and the drug concentration. Observed frequency- and concentration-dependent blocks were consistent with the predictions of the model, and mixture-specific rate constants were estimated from these data. The negligible shift in channel inactivation and the reduction of apparent binding and unbinding rates with more polarized membrane potentials imply the active moiety of ethacizin blocks open channels and is trapped within the channel at resting membrane potentials. The binding rate at 0 mV is similar to that observed in studies of interactions of other open channel blocking agents with voltage- and ligand-gated channels.

Transient outward current in catecholamine-induced cardiac hypertrophy in the rat

1996 ◽  
Vol 271 (6) ◽  
pp. H2360-H2367 ◽  
Author(s):  
J. Meszaros ◽  
K. O. Ryder ◽  
G. Hart
Keyword(s):  
Cardiac Hypertrophy ◽  
Weight Ratio ◽  
Outward Current ◽  
Cell Voltage ◽  
Underlying Mechanism ◽  
Left Ventricular ◽  
Daily Injection ◽  
Heart Weight ◽  
Inactivation Kinetics ◽  
Transient Outward Current

We have demonstrated that a daily injection of isoproterenol (5 mg/kg ip) for 7 days induces a 30% increment in heart weight-to-body weight ratio and prolongs the action potential duration (APD) in male Wistar rats. The underlying mechanism of the prolonged APD was investigated in this model of hypertrophy by measuring the transient outward potassium current (Ito) in left ventricular myocytes of the rat with whole cell voltage-clamp techniques. Cell membrane capacitance was increased by 39%: 122 +/- 3 (n = 23) and 171 +/- 5 (SE) pF (n = 20) in control and hypertrophy, respectively (P < 0.001). Ito was evoked in sodium-free solutions containing 0.5 mM Ca2+ and 2 mM Co2+ by step depolarizations from a holding potential of -80 mV. The amplitude of the 4-aminopyridine-sensitive Ito (at 70 mV) was reduced by 28% in hypertrophy: 3.2 +/- 0.3 (n = 23) and 2.3 +/- 0.4 (SE) nA (n = 20) in control and hypertrophy, respectively (P < 0.05). When normalized for cell capacitance, the reduction was much larger: 26.4 +/- 2.5 and 13.1 +/- 1.8 pA/pF in control and hypertrophy, respectively (P < 0.001). The voltage dependence of Ito was similar in both cell types. No change was observed in the steady-state activation and inactivation kinetics in the two groups, nor was there a change in the time dependence of inactivation. The recovery from inactivation of Ito when fitted with a monoexponential function was not changed significantly in hypertrophy: time constants = 8.2 +/- 0.4 (n = 13) and 8.3 +/- 0.3 ms (n = 12) in control and hypertrophy, respectively. These results show that Ito density is decreased in catecholamine-induced cardiac hypertrophy, but current kinetics are not affected. The reduced Ito density may underlie the prolongation of APD in this model of hypertrophy.

Role of chloride currents in repolarizing rabbit atrial myocytes

1995 ◽  
Vol 268 (5) ◽  
pp. H1992-H2002 ◽  
Author(s):  
Z. Wang ◽  
B. Fermini ◽  
J. Feng ◽  
S. Nattel
Keyword(s):  
Action Potential ◽  
Reversal Potential ◽  
Outward Current ◽  
Cell Voltage ◽  
Disulfonic Acid ◽  
Transient Outward Current ◽  
Atrial Myocytes ◽  
Atrial Cells ◽  
K Current

Rabbit atrial cells manifest a prominent transient outward K+ current (Ito1), but this current recovers slowly from inactivation and is unlikely to be important at physiological rates (3-5 Hz). Depolarization of rabbit atrial cells also elicits a transient Ca(2+)-dependent outward Cl- current (Ito2). To compare the relative magnitude of these transient outward currents at various rates, we applied whole cell voltage-clamp techniques to isolated rabbit atrial myocytes. Whereas peak Ito1 exceeded Ito2 at slow rates (0.1 Hz), Ito1 was strongly reduced as rate was increased (by 97 +/- 2%, mean +/- SE, at 4 Hz), while Ito2 was slightly reduced (by 28 +/- 4%, 4 Hz). The reversal potential of transient outward tail currents at 0.07 Hz was -49 +/- 9 mV, while at 2.5 Hz the reversal potential became -18 +/- 7 mV (calculated Cl- reversal potential -18 mV). The addition of the Cl- transport blocker 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS; 150 microM) or the replacement of external Cl- with methanesulfonate inhibited a large part of the transient outward current elicited by depolarization at 4 Hz. DIDS and Cl- replacement increased action potential duration in both single rabbit atrial cells and multicellular rabbit atrial preparations. We conclude that the Ca(2+)-dependent Cl- current is substantially larger than the transient K+ current at physiological rates in the rabbit and is likely to play a more important role in action potential repolarization than the latter current in this tissue in vivo.

Activators of protein kinase C decrease Ca2+ spark frequency in smooth muscle cells from cerebral arteries

1997 ◽  
Vol 273 (6) ◽  
pp. C2090-C2095 ◽  
Author(s):  
Adrian D. Bonev ◽  
Jonathan H. Jaggar ◽  
Michael Rubart ◽  
Mark T. Nelson
Keyword(s):  
Protein Kinase C ◽  
Membrane Potential ◽  
Protein Kinase ◽  
Cerebral Arteries ◽  
Outward Current ◽  
Transient Outward Current ◽  
Open Probability ◽  
Kinase C ◽  
Voltage Dependent ◽  
Channel Currents

Local Ca2+ transients (“Ca2+ sparks”) caused by the opening of one or the coordinated opening of a number of tightly clustered ryanodine-sensitive Ca2+-release (RyR) channels in the sarcoplasmic reticulum (SR) activate nearby Ca2+-dependent K+(KCa) channels to cause an outward current [referred to as a “spontaneous transient outward current” (STOC)]. These KCa currents cause membrane potential hyperpolarization of arterial myocytes, which would lead to vasodilation through decreasing Ca2+ entry through voltage-dependent Ca2+ channels. Therefore, modulation of Ca2+spark frequency should be a means to regulation of KCa channel currents and hence membrane potential. We examined the frequency modulation of Ca2+ sparks and STOCs by activation of protein kinase C (PKC). The PKC activators, phorbol 12-myristate 13-acetate (PMA; 10 nM) and 1,2-dioctanoyl- sn-glycerol (1 μM), decreased Ca2+ spark frequency by 72% and 60%, respectively, and PMA reduced STOC frequency by 83%. PMA also decreased STOC amplitude by 22%, which could be explained by an observed reduction (29%) in KCa channel open probability in the absence of Ca2+ sparks. The reduction in STOC frequency occurred in the presence of an inorganic blocker (Cd2+) of voltage-dependent Ca2+ channels. The reduction in Ca2+ spark frequency did not result from SR Ca2+ depletion, since caffeine-induced Ca2+ transients did not decrease in the presence of PMA. These results suggest that activators of PKC can modulate the frequency of Ca2+ sparks, through an effect on the RyR channel, which would decrease STOC frequency (i.e., KCa channel activity).

Characterization of macroscopic outward currents of canine colonic myocytes

1989 ◽  
Vol 257 (3) ◽  
pp. C461-C469 ◽  
Author(s):  
W. C. Cole ◽  
K. M. Sanders
Keyword(s):  
Outward Current ◽  
Cell Voltage ◽  
Muscle Cells ◽  
Time Dependent ◽  
Delayed Rectifier ◽  
Delayed Rectifier Current ◽  
Outward Currents ◽  
Voltage Dependent ◽  
K Current ◽  
Time Courses

Outward currents of colonic smooth muscle cells were characterized by the whole cell voltage-clamp method. Four components of outward current were identified: a time-independent and three time-dependent components. The time-dependent current showed strong outward rectification positive to -25 mV and was blocked by tetraethylammonium. The time-dependent components were separated on the basis of their time courses, voltage dependence, and pharmacological sensitivities. They are as follows. 1) A Ca2+-activated K current sensitive to external Ca2+ and Ca2+ influx was blocked by ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (0.1 X 10(-3) M) and nifedipine (1 X 10(-6) and was increased by elevated Ca2+ (8 X 10(-6) M) and BAY K 8644 (1 X 10(-6) M). 2) A "delayed rectifier" current was observed that decayed slowly with time and showed no voltage-dependent inactivation. 3) Spontaneous transient outward currents that were blocked by ryanodine (2 X 10(-6) M) were also recorded. The possible contributions of these currents to the electrical activity of colonic muscle cells in situ are discussed. Ca2+-activated K current may contribute a significant conductance to the repolarizing phase of electrical slow waves.

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