Responses of the Transmembrane Potential of Myocardial Cells during a Shocks

Responses of Transmembrane Potential During a Shock. Introduction: The purpose of this investigation was to study the transmembrane potential changes (δV_m) during extracellular electrical field stimulation. Methods and Results: V_m was recorded in seven guinea pig papillary muscles in a tissue hath by a double-barrel microelectrode with one barrel in and the other just outside a cell while shocks were given across the bath. The short distance (15 to 30 μm) between the two microelectrode tips and alignment of the tips parallel to the shock electrodes eliminated the shock artifact. Following ten SI stimuli, an S2 shock field created by a 10-msec square wave was delivered during the action potential plateau or during diastole through shock electrodes 1 cm on either side of the tissue. Four shock strengths creating field strengths of 1.7 ± 0.1, 2.9 ± 0.2, 6.1 ± 0.6. and 8.8 ± 0.9 V/cm were given for the same impalement. Both shock polarities were given at each shock strength. For shocks delivered during the action potential plateau, the magnitudes of the peak δV_m caused by the above four potential gradients were 21.1 ± 8.2, 33.6 ± 13.6, 49.9 ± 24.2, and 52.3 ± 28.0 mV (P < 0.05 among the four groups) for the shocks causing depolarization and 37.9 ± 14.2,56.6 ± 16.4,83.1 ± 19.4. and 92.9 ± 29.1 mV (P < 0.05 among the four groups) for the shocks causing hyperpolarization. Though δV_m increased as potential gradients increased, the relationship was not linear. The magnitude of hyperpolarization was 1.9 ± 0.5 times that of depolarization when the shock polarity was reversed (P < 0.05). As potential gradients increased from 1.7 ± 0.1 to 8.8 ± 0.9 V/cm, the time constant of the membrane response decreased significantly from 3.5 ± 1.8 to 1.6 ± 0.7 msec for depolarizing shocks and from 6.0 ± 3.1 to 3.4 ± 1.9 msec for hyperpolarizing shocks (P < 0.01 vs depolarizing shocks). For shocks delivered during diastole, hyperpolarizing shocks induced triphasic changes in V_m during the shock, i.e., initial hyperpolarization, then depolarization, followed again by hyperpolarization. Conclusion: During the action potential plateau, the membrane response cannot be represented by a classic passive RC membrane model. During diastole, activation upstrokes occur even during hyperpolarization caused by shocks creating potential gradients between aproximately 2 and 9 V/cm.     

Prolongation of repolarization time by electric field stimulation with monophasic and biphasic shocks in open-chest dogs

Recent studies suggest that 1) electrically induced fibrillation and defibrillation involve prolongation of refractoriness by the shock in addition to stimulation and 2) biphasic waveforms are more efficient for defibrillation than are comparable monophasic waveforms. The purpose of this study was to compare prolongation of action potential duration at 50% repolarization by monophasic and biphasic shocks during paced rhythm. A floating glass microelectrode was used to record intracellularly from the anterior right ventricular epicardium in seven open-chest dogs. After 10 S1 beats paced at an interval of 350 msec, 5-msec and 2.5-msec monophasic shocks and biphasic shocks, with each phase of 2.5 msec, were given via mesh electrodes on either side of the microelectrode. The shock strength was adjusted so that the shock field, measured from eight extracellular electrodes encircling the microelectrode, was about 5 V/cm. Monophasic and biphasic S2 shocks were given starting with an S1-S2 interval of 120 msec, which was increased in 5-msec steps until an action potential was produced by the S2 shock. Both monophasic and biphasic 5 V/cm shock fields caused significant prolongation of action potential duration. The prolongation of action potential duration increased as the S1-S2 interval increased. This prolongation occurred at shorter S1-S2 intervals for 5-msec monophasic shocks than for biphasic shocks.     

Pertussis toxin treatment blocks hyperpolarization by muscarinic agonists in chick atrium

Atrial and ventricular adenylate cyclase activity and atrial membrane potentials were measured in hearts from hatched chicks at 2-3 days after intravenous administration of pertussis toxin (0.5-1.0 micrograms, total) or saline. Both in atrium and ventricle, treatment with pertussis toxin antagonized inhibition by carbachol of basal and isoproterenol-stimulated adenylate cyclase activity without changing either basal or isoproterenol-stimulated adenylate cyclase. In atria from pertussis toxin-treated animals (5.4 mM potassium), carbachol hyperpolarized the resting membrane by 0.3 +/- 0.3 mV (n = 9) and did not increase resting potassium conductance. In contrast, carbachol hyperpolarized the resting membrane by 4.5 +/- 0.8 mV (n = 11) and increased resting potassium conductance more than 4-fold in saline-treated animals. Carbachol did not significantly affect the atrial action potential peak or duration at 50% repolarization of pertussis toxin-treated animals. This muscarinic agonist reduced action potential peak by 7.8 +/- 1.2 mV and the duration at 50% repolarization by 22.1 +/- 3.0 msec in atria from saline-treated animals. Pertussis toxin treatment also prevented the negative inotropic effect and the inhibition of calcium-dependent action potentials caused by carbachol in atrial muscle. Neither the affinity nor the maximal specific binding of [3H]quinuclidinyl benzilate in ventricular homogenates was changed by pertussis toxin treatment. The apparent affinity of carbachol for muscarinic receptor was slightly (approximately 2-fold) diminished in pertussis toxin-treated animals. The inhibition of carbachol-induced hyperpolarization by pertussis toxin treatment implicates a guanosine 5'-triphosphate-dependent protein (Ni or a similar protein) as an essential link that permits muscarinic receptor to regulate atrial potassium channels.     

Nonlinear changes of transmembrane potential during electrical shocks: role of membrane electroporation

Defibrillation shocks induce nonlinear changes of transmembrane potential (DeltaVm) that determine the outcome of defibrillation. As shown earlier, strong shocks applied during action potential plateau cause nonmonotonic negative DeltaVm, where an initial hyperpolarization is followed by Vm shift to a more positive level. The biphasic negative DeltaVm can be attributable to (1) an inward ionic current or (2) membrane electroporation. These hypotheses were tested in cell cultures by measuring the effects of ionic channel blockers on DeltaVm and measuring uptake of membrane-impermeable dye. Experiments were performed in cell strands (width approximately 0.8 mm) produced using a technique of patterned cell growth. Uniform-field shocks were applied during the action potential plateau, and DeltaVm was measured by optical mapping. Shock-induced negative DeltaVm exhibited a biphasic shape starting at a shock strength of approximately 15 V/cm when estimated peak DeltaV-m was approximately -180 mV; positive DeltaVm remained monophasic. Application of a series of shocks with a strength of 23+/-1 V/cm resulted in uptake of membrane-impermeable dye propidium iodide. Dye uptake was restricted to the anodal side of strands with the largest negative DeltaVm, indicating the occurrence of membrane electroporation at these locations. The occurrence of biphasic negative DeltaVm was also paralleled with after-shock elevation of diastolic Vm. Inhibition of I(f) and I(K1) currents that are active at large negative potentials by CsCl and BaCl2, respectively, did not affect DeltaVm, indicating that these currents were not responsible for biphasic DeltaVm. These results provide evidence that the biphasic shape of DeltaVm at sites of shock-induced hyperpolarization is caused by membrane electroporation.     

Intracellular studies of electrical membrane properties of opossum esophageal circular smooth muscle

It has been suggested that regional differences in membrane properties of circular esophageal smooth muscle play an important role in the mechanism of esophageal peristalsis. The purpose of this study was to examine both the passive and active membrane properties of circular smooth muscle at proximal and distal esophageal sites so as to delineate the role of myogenic properties in the intramural mechanism of peristalsis. Intracellular recordings were made in circular muscle strips taken from proximal (8 cm above the gastroesophageal junction) and distal (2 cm above the gastroesophageal junction) sites in 10 opossums using the partition method of Abe and Tomita. At both esophageal sites, determinations were made of resting membrane potentials, time constants, space constants, thresholds for action potentials, action potential amplitudes, rates of rise of action potentials, and action potential durations at half-amplitude. The values for these parameters at the proximal and distal sites, respectively, were as follows: mean resting membrane potential, 49.7 +/- 0.24 and 49.5 +/- 0.3 mV; length constant, 4.0 +/- 0.4 and 3.8 +/- 0.4 mm; time constant, 513 +/- 49 and 456 +/- 53 ms; threshold for action potentials, 9.3 +/- 0.4 and 8.8 +/- 0.3 mV; amplitude of action potentials, 36.0 +/- 5.2 and 35.3 +/- 1.7 mV; rate of rise of action potentials, 2.3 +/- 0.3 and 2.6 +/- 0.4 mV/ms; duration of action potentials at half-amplitude, 5.0 +/- 1.2 and 4.1 +/- 0.4 ms; and the conduction velocity for evoked potentials, 3.9 +/- 0.3 and 3.8 +/- 0.4 cm/s. Our studies show that there are no differences between proximal and distal esophageal sites in any of these determinations. These studies also show that regional differences in the electrical membrane properties of circular smooth muscle do not account for esophageal peristalsis.     

Hyperpolarization of the membrane potential caused by somatostatin in dissociated human pituitary adenoma cells that secrete growth hormone.

Membrane electrical properties and the response to somatostatin were examined in dissociated human pituitary adenoma cells that secrete growth hormone (GH). Under current clamp condition with a patch electrode, the resting potential was -52.4 +/- 8.0 mV, and spontaneous action potentials were observed in 58% of the cells. Under voltage clamp condition an outward K+ current, a tetrodotoxin-sensitive Na+ current, and a Ca2+ current were observed. Cobalt ions suppressed the Ca2+ current. The threshold of Ca2+ current activation was about -60 mV. Somatostatin elicited a membrane hyperpolarization associated with increased membrane permeability in these cells. The reversal potential of somatostatin-induced hyperpolarization was -78.4 +/- 4.3 mV in 6 mM K+ medium and -97.2 +/- 6.4 mV in 3 mM K+ medium. These reversal potential values and a shift with the external K+ concentration indicated that membrane hyperpolarization was caused by increased permeability to K+. The hyperpolarized membrane potential induced by somatostatin was -63.6 +/- 5.9 mV in the standard medium. This level was subthreshold for Ca2+ and Na+ currents and was sufficient to inhibit spontaneous action potentials. Hormone secretion was significantly suppressed by somatostatin and cobalt ions. Therefore, we suggest that Ca2+ entering the cell through voltage-dependent channels are playing an important role for GH secretion and that somatostatin suppresses GH secretion by blocking Ca2+ currents. Finally, we discuss other possibilities for the inhibitory effect of somatostatin on GH secretion.     

Diabetic state-induced modification of resting membrane potential and conductance in diaphragm muscle of alloxan and diabetic KK-CAy mice

Summary The electrical properties of skeletal muscle membranes were investigated in genetically diabetic KK-CA^y mice and alloxan-induced diabetic ddY mice. Using isolated phrenic nerve-diaphragm muscle or sciatic nerve-gastrocnemius muscle in situ preparations, nerve-stimulated twitch tensions (the maximal value) were obtained at lower voltage pulse in diabetic KK-CA^y mice than in normal ddY mice. The diabetic state reduced resting membrane potentials (1.7–4.0 mV) and resting membrane conductance (0.37–0.44 siemen), decreased the amplitude (3.8–3.9 mV) and overshoot (4.5 mV) of directly induced-action potential, and prolonged action potential duration. In the diabetic state, resting membrane conductance was multiply-correlated with blood glucose level and resting membrane potential. In alloxan-induced diabetic mice, resting membrane potentials were significantly multiply-correlated with the weeks elapsed after alloxan injection and blood glucose level (p<0.01). Since the reduction of resting membrane potential correlated with the weeks, changes in resting membrane potential may be involved in the decrease in insulin-like growth factor action. The reduction of resting membrane conductance was correlated with the increase in blood glucose.     

Frequency and voltage dependent effects of AN-132, a new class I anti-arrhythmic agent, on Vmax of action potential upstroke in guinea pig ventricular muscles

The in vitro electrophysiological properties of a newly synthesised antiarrhythmic agent, AN-132, were evaluated by recording transmembrane action potentials from guinea pig papillary muscles. AN-132 (10-100 mumol.litre-1) caused a dose dependent decrease in the maximum upstroke velocity (Vmax) of the action potential without affecting the resting potential. In the presence of AN-132, trains of stimuli at rates greater than or equal to 0.1 Hz led to an exponential decline in Vmax. This use dependent block was enhanced at higher stimulation frequency. The time constant for the recovery of Vmax from the use dependent block was 39.5-41.2 s. The curves relating membrane potential and Vmax were shifted by AN-132 (100 mumol.litre-1) in the direction of more negative potentials (6.1 mV). In preparations treated with AN-132 (30 and 100 mumol.litre-1), the Vmax of test action potentials preceded by conditioning clamp pulses to 0 mV was progressively decreased with an increasing number of pulses. A single prolonged clamp pulse to 0 mV reduced Vmax much less than multiple brief clamp pulses. These findings suggest than AN-132 has use dependent inhibitory action on the fast sodium channel by binding to the channel mainly during its activated state and that the unbinding rate of the drug during diastole is very slow. This use dependency and its greater inhibition of Vmax in depolarised muscles through the increase in tonic block may play a major role in preventing ventricular arrhythmias.     

Restorative effect of epinephrine on the electrophysiologic properties of depressed human atrial tissue

The effects of epinephrine on the electrophysiologic properties of human right atrial tissue, obtained at cardiac surgery, were evaluated utilizing standard microelectrode techniques. In studies of electrophysiology, epinephrine had little effect on the resting membrane potential and transmembrane action potentials of normal atrial fibers. Epinephrine enhanced phase-4 depolarization and increased automaticity in normal fibers but hyperpolarized partially depolarized atrial fibers and decreased automaticity. The hyperpolarizing action of epinephrine resulted in a increase in action potential amplitude and dV/dt and enhanced conduction. Active force increased 40–230% in depressed tissues when exposed to epinephrine. Epinephrine-induced hyperpolarization of depressed atrial fibers may have a beneficial effect on atrial arrhythmias and depressed contractility encountered clinically.     

Usefulness of the Evaluation of Isovolumic and Ejection Phase Myocardial Signals during Stress Echocardiography in Predicting Exercise Capacity in Heart Failure Patients

Aim: To assess changes of systolic function using tissue Doppler imaging (TDI) during stress echocardiography and its impact on exercise capacity in heart failure (HF) patients (pts). Material and Methods: 80 pts (65 male), mean age of 59.3 ± 10.9 years, NYHA class 1.95 ± 0.8, left ventricle ejection fraction (LVEF) 27.2 ± 9.5 (10−45%). The etiology of HF was ischemic (ICM) in 50 pts and dilated cardiomyopathy (DCM) in 30 pts. Peak myocardial velocity (IVV) and acceleration (IVA) during isovolumic contraction and peak myocardial velocity during ejection phase (S') were measured at baseline and peak exercise during semi-supine stress-echo (20 Watts, 2-min increments). Concurrently peak oxygen uptake (VO₂ peak) was measured. Results: Rest values of analyzed parameters were comparable in groups according to etiology of HF and physical capacity. However, peak stress parameters mainly S' were significantly higher in the DCM group and the group with better VO₂ peak. The best correlation with exercise capacity was S' at peak stress (r = 0.66; p < 0.0001). The most useful parameter for identifying severe exercise intolerance, VO₂ peak < 14 ml/kg/min, was S' with an area under ROC curve of 0.82 ± 0.05 (95% CI 0.71−0.89). The cutoff of 5.75 cm/s for S' at peak stress showed a sensitivity of 61% with a specificity of 96%. Conclusions: The evaluation of systolic function by means of TDI instead of LVEF shows more clearly that systolic function is at least partly responsible for exercise tolerance in HF. Assessment of echocardiographic systolic parameters at peak stress provides more accurate information about exercise capacity in HF pts.     

Hypertrophic smooth muscle in the partially obstructed opossum esophagus. Excitability and electrophysiological properties

Partial obstruction of the opossum esophagus leads to thickening of the circular muscle, hypertrophy of smooth muscle cells, and diminution of the extracellular space. The pharmacological and electrophysiological properties of this hypertrophied muscle were studied. Carbachol produced phasic and tonic contractions of the circular muscle. The EC50 for tonic contractions was greater for hypertrophied than for normal muscle (21.1 +/- 3.9 mumol/L vs. 4.8 +/- 2.2 mumol/L; P less than 0.05). The resting membrane potential difference of hypertrophied muscle (-50.8 +/- 0.2 mV) was similar to that of normal muscle (-50.0 +/- 0.2 mV). Electrical stimulation of intrinsic nerves in the normal muscle produced a hyperpolarization followed by a depolarization of smooth muscle membrane potential. Hypertrophied muscle responded either with an attenuated hyperpolarization or no hyperpolarization, both of which were followed by a depolarization. The space constant in the long axes of the hypertrophied circular muscle cells was greater than normal (4.4 +/- 0.2 mm vs. 3.4 +/- 0.1 mm; P less than 0.001). The threshold potential for initiation of action potentials was more negative for hypertrophied (-43.2 +/- 0.4 mV) than for normal circular muscle (-41.6 +/- 0.2 mV; P less than 0.005). These data indicate that alterations in neuromuscular function accompany the hypertrophy of esophageal smooth muscle.     

Myasthenic syndrome caused by mutation of the SCN4A sodium channel

In a myasthenic syndrome associated with fatigable generalized weakness and recurrent attacks of respiratory and bulbar paralysis since birth, nerve stimulation at physiologic rates rapidly decremented the compound muscle action potential. Intercostal muscle studies revealed no abnormality of the resting membrane potential, evoked quantal release, synaptic potentials, acetylcholine receptor channel kinetics, or endplate ultrastructure, but endplate potentials depolarizing the resting potential to -40 mV failed to excite action potentials. Pursuing this clue, we sequenced SCN4A encoding the skeletal muscle sodium channel (Nav1.4) and detected two heteroallelic mutations involving conserved residues not present in 400 normal alleles: S246L in the S4/S5 cytoplasmic linker in domain I, and V1442E in the S3/S4 extracellular linker in domain IV. The genetically engineered V1442E-Na channel expressed in HEK cells shows marked enhancement of fast inactivation close to the resting potential, and enhanced use-dependent inactivation on high-frequency stimulation; S246L is likely a benign polymorphism. The V1442E mutation in SCN4A defines a novel disease mechanism and a novel phenotype with myasthenic features.     

Evaluation of Longitudinal Tissue Velocity and Deformation Imaging in Akinetic Nonviable Inferobasal Segments of Left Ventricular Myocardium by Dobutamine Stress Echocardiography

Aim: To study tissue velocity imaging (TVI) and strain rate imaging (SRI) indices in akinetic nonviable and normal left ventricular (LV) inferobasal segment and effect of dobutamine infusion on these indices in nonviable segments. Methods: The study population consisted of two groups: 25 patients (mean age 60.75 ± 8.69 years) with left ventricular akinetic inferobasal nonviable segment determined by dobutamine stress echocardiography (DSE) and 14 normal coronaries (mean age 56.67 ± 11.90 years) with normal echocardiography as control group. The following TVI and SRI parameters were measured in patient and control group: ejection phase velocity (Sm [cm/sec]), peak systolic strain (ST [%]), and strain rate (SR [per second])). Results: Ejection fraction was significantly lower in patient group (29.40%± 5.46% vs. 55.00%± 3.39%; P < 0.001). Several differences were observed in patients with nonviable inferobasal segments compared to control group: Sm was reduced (3.58 ± 1.08 cm/sec vs. 5.56 ± 1.28 cm/sec; P < 0.001); SR and ST were significantly decreased (−0.39 ± 0.20/second vs. −1.44 ± 0.64/second, and −3.86%± 4.12% vs. −17.64%± 7.44%, respectively; P < 0.001 in both). The range of SR for nonviable segments (−0.04 to −0.77/second) did not overlap with that of the normal segments (−0.80 to −3.0/second). This range for Sm and ST overlapped with those of the normal segments. Conclusion: All TVI and SRI parameters are reduced in akinetic nonviable inferobasal compared with normal segments. According to findings of this study, resting strain rate has a potential to discriminate nonviable inferobasal from normal segments.     

Junction Potentials and Action Potentials of the Crayfish Myocardium

Junction and Action Potentials. Introduction : The purpose of this investigation was to study the properties of junction and action potentials elicited by nerve stimulation in the absence of the cardiac ganglion in crayfish myocardium.
Methods and Results : The cardiac ganglion was surgically removed in isolated crayfish hearts. Electrical stimulation of one of the peripheral anterolateral nerves provided isolated junction potentials and action potentials free of the usual postspike bursts of junction potentials. The single junction potentials displayed amplitudes of up to 25 mV and slow exponential decay; the mean time constant was 170 ± 13 msec (xT ± SD). In fully recovered tissue, the junction potentials triggered action potentials free of repetitive subthreshold discharges. Tetrodotoxin did not alter the amplitude or shape of action potentials initiated by direct electrical stimulation of the muscle cells. Calcium channel blocking agents such as Cd²⁺ and Ni²⁺ eliminated the action potentials but not the junction potentials. Tetraethylammonium markedly prolonged the action potential duration.
Conclusions : Our data suggest that: (1) A slow decay of the junction potentials may result from the disappearance of the neurotransmitter; this process also accounts for the late slow repolarization of the final part of the action potentials; (2) The equilibrium potential of the junction potentials is close to 0 mV; (3) The upstroke of the action potentials is carried by calcium currents; (4) The fast repolarization phase of the action potentials is likely caused by the delayed rectifier; and (5) The refractory phase outlasts the duration of the action potential.     

Ouabain-resistant hyperpolarization induced by insulin in aggregates of embryonic heart cells.

Spheroidal aggregates formed from trypsin-dissociated 14-day embryonic chicken hearts after 48 hr of rotation on a gyratory shaker. Intracellularly recorded resting membrane potentials of aggregates bathed in 1.3 mM K+ balanced salt solution had a mean (+/- SD) of 64 +/- 4 mV. After a stable potential was achieved, addition of 1-100 nM sodium bovine insulin caused a slow hyperpolarization of up to 19 mV after 4-5 min, followed, in some cases, by a further, more rapid, shift to a potential near EK. Equivalent hyperpolarizations were observed when insulin was added in the presence of 10 mM ouabain, indicating that enhanced Na+,K+ pump activity was not responsible for the change in membrane potential. The concentration of insulin that produced half-maximal hyperpolarization (2 nM) corresponded to the association constant of a high-affinity insulin receptor, suggesting that binding to this class of receptors led to the change in membrane potential. Steady-state current-voltage curves from current clamp experiments suggested that insulin produced an increase in slope conductance at potentials near rest by inducing an outward current with an apparent potential negative to -90 mV.     

Fragmented, long-duration, low-amplitude electrograms characterize the origin of focal atrial tachycardia

Background: Focal atrial tachycardias (FAT) originate from areas with poor cell-to-cell coupling. Due to cellular uncoupling extracellular potentials become fractionated. The degree of fragmentation may be used to identify the site of origin of FAT prior to catheter ablation. We studied electrical fragmentation in relation to the distance to the site of earliest activity during FAT.
Methods: Three-dimensional (3-D) electroanatomical activation/voltage maps obtained from patients (n = 15: 6 male, age 40 ± 14 (19–72) years) referred for catheter ablation of FAT were analyzed. Bipolar atrial potentials (BP) were categorized according to the number of deflections. The peak-to-peak amplitude and the time interval between the first and last deflection ( fractionation duration) of each BP were measured.
Results: Eighteen different atrial tachycardias (AT) (CL 346 ± 109 [190–550]) msec were analyzed. The incidence of single potentials ranged from 30 to 81 (59 ± 16)%. The occurrence of double and fractionated potentials ≤2 cm of the site of earliest activity ("focal area") was higher compared to the remainder of the atria (67 ± 22% vs 34 ± 14%, P < 0.001). Focal area potentials were characterized by a longer duration (49 ± 22 msec vs 34 ± 17 msec, P < 0.001) and a lower peak-to-peak amplitude (0.51 ± 0.43 [0.12–1.7] mV vs 0.94 ± 0.69 [0.22–2.58] mV, P = 0.03). Fractionation was not associated with FAT cycle length (r =−0.26, P = 0.29) or left atrial diameter (r = 0.47, P = 0.30).
Conclusion: Significant differences in fractionation, fractionation duration, and peak-to-peak-amplitude of atrial potentials between the focal area and the remainder of the atria exist. Fractionation and voltage mapping can be used, besides activation mapping, to identify the site of origin of FAT during catheter ablation.     

Sodium channels induced by depolarization of the Xenopus laevis oocyte.

An electrically gated Na+ channel can be made to appear in the membrane of the Xenopus laevis oocyte by simple depolarization. This membrane normally responds passively to imposed transmembrane currents with resting potentials around -60 mV, but when it is held depolarized to more than about +30 mV it becomes possible to obtain long-lasting regenerative depolarizations up to +80 mV; these depolarizations can last as long as 20 min. This potential is due to an "induction" of a Na+-dependent channel that is electrically gated open and closed. Its threshold for opening is about -20 mV and it is selective for Na+ over Cs+ and choline+ but is blocked by relatively small quantities of Li+. When a long voltage clamp step to a positive potential under ENa (+70 to +90 mV) is applied, an inward current is observed for many minutes, implying that this channel does not have an inactivation mechanism. The inward Na+ current is blocked by 0.50 mM tetrodotoxin. When the membrane is held at or near resting potential, the excitability will disappear with time, but it can be made to reappear by again depolarizing the membrane.     

Elevations in arterial pressure induce the formation of spontaneous action potentials and alter neurotransmission in canine ileum arteries

Arterial segments (less than 250 micron o.d.) excised from canine ileum were mounted in a chamber that permitted arterial transmural pressure (TMP) to be altered and measured. Subsequently, the periarterial nerves were field stimulated with single pulses (0.1 msec, 70 V), and the resting membrane potential (Em) as well as the nerve-mediated alterations in smooth muscle Em were measured using intracellular microelectrodes at TMPs between 0 and 160 mm Hg. The resting Em was greatest at TMPs of 40 mm Hg (-54.7 +/- 2.6 mV) and depolarized as the TMP was increased, reaching a value of -44.8 +/- 3.1 mV at 160 mm Hg. At TMP greater than or equal to 60 mm Hg, a proportion of the preparations exhibited spontaneous electrical activity (SA) consisting of constant rhythmic oscillations in Em or action potentials (APs) or of trains of rhythmic APs that progressively decreased in amplitude, interrupted by periods of hyperpolarization. SA stopped when the TMP was lowered to 40 mm Hg and was reestablished when the TMP was reelevated to TMPs above 60 mm Hg. Nerve stimulation evoked excitatory junction potentials (ejps) or APs. At constant stimulus parameters, ejps of maximum amplitude having the greatest rate of potential rise and fall were produced at TMP of 100 mm Hg. At TMPs greater than 100 mm Hg or less than 100 mm Hg, the amplitude and the rate of rise and fall of the ejps decreased. Ejps formed in response to a constant single pulse stimulus (0.1 msec, 70 V) elicited APs only at TMPs greater than or equal to 60 mm Hg. Neither ejps nor APs were inhibited by alpha-receptor-blocking agents. These studies indicate that the TMP at which an artery is maintained plays an important role in determining the resting Em, the occurrence of spontaneous action potentials, and the alterations in Em associated with nerve stimulation.     

Ventricular Action Potential and L-Type Calcium Channel in Infarct-Induced Hypertrophy in Rats

Calcium Channel in Infarct-Induced Hypertrophied Rat Ventricle. Introduction : The present investigation was aimed at characterization of: (1) action potential parameters; and (2) L-type calcium channels in the hypertrophied ventricular tissue surviving an extensive healed myocardial infarction in the rat.
Methods and Results : Myocardial infarction was produced in Wistar rats by ligation of the left coronary artery. One to 2 months later, their hearts were subjected to electrophysiologic study. The main difference in subendocardial transmembrane potentials recorded with intracellular microelectrodes was an increase in action potential duration (APD). In the left ventricle, the infarcted/sham-operated APD ratio ranged from 2.7 to 7.2, whereas in the right ventricle it ranged from 1.6 to 2.3 in different regions. When compared with control ceils, ventricular myocytes from infarcted hearts were found to be larger (P < 0.01) and showed a reduction (P < 0.05) in L-type calcium current (I_Ca,L) density obtained by whole cell, patch clamp (at 0 mV: 4.44 ± 0.41 in infarcted vs 8.03 ± 1.22 pA/pF in normal). The time course of decay of the currents could be fitted by two exponential functions in both normal and infarcted hearts. There was a tendency toward an increase in the time constant of the slower component of inactivation, T₂, significant only at +20 mV (215 ± 25 vs 151 ± 15 msec).
Conclusions : Cardiac hypertrophy of healed infarction in rats is associated with lengthening of the action potential in both ventricles. The main alteration observed in I^Ca,Lwas a decrease in the current density. Thus, alteration of the calcium channel is not the determinant factor of APD increase