Potassium Contracture Research Articles

Calcium release in skinned muscle fibres of the toad by transverse tubule depolarization or by direct stimulation.

1. Skeletal muscle fibres from the toad were mechanically skinned under paraffin oil and then bathed in a potassium HDTA solution (HDTA: hexamethylenediamine-tetraacetate) which mimicked the ionic composition of the myoplasm. 2. Rapid transient contractions could be triggered by substitution of K+ with Na+ (with no change of anion), which should have virtually no direct effect on the electrical polarization of the sarcoplasmic reticulum (SR) membrane. Up to thirty or more contractions could be evoked by repeated substitutions if there was sufficient 'repriming' time (about 30 s) between them; these rapid contractions were analagous to potassium contractures in intact fibres. 3. When the SR was not heavily loaded, substitution of potassium HDTA with choline chloride also produced a rapid, brief contraction. 4. All treatments designed to 'inactivate' the voltage sensor in the T-system invariably abolished the rapid contractions. Thus, rapid contractions were absent if (i) the T-system was permanently depolarized by pre-soaking the muscle in a high potassium solution with ouabain before skinning, (ii) a fibre was split rather than skinned, (iii) the T-system was temporarily depolarized by Na+ substitution immediately before choline chloride substitution, or vice versa, (iv) a skinned fibre was briefly exposed to saponin (50 micrograms/ml) to selectively disrupt the T-system membrane or (v) the muscle was pre-soaked in a solution with 1 mM-EGTA and no Ca2+ or Mg2+ before skinning. In contrast to (v), if 10 mM-Mg2+ was present in the EGTA solution before skinning, rapid contractions could be elicited, presumably because the presence of Mg2+ prevented the inactivation of the T-system voltage sensor in low [Ca2+]. 5. These results unequivocally demonstrate that (a) the T-system reseals and repolarizes after mechanical skinning under oil and (b) the fast contractions are produced by activation of the voltage sensor in the T-system. 6. When the SR had been heavily loaded, choline chloride substitution (but not Na+ substitution) could also induce an unphysiological, slow contraction ('second component'). In total contrast to the fast contraction, this slow component was unaffected by any of the treatments (i-v) above, indicating that it did not depend on activation of the voltage sensor in the T-system but resulted from a direct action of choline chloride on the SR.(ABSTRACT TRUNCATED AT 400 WORDS)

Dihydropyridine calcium channel antagonists block and agonists potentiate high potassium contractures but not twitches in frog skeletal muscle.

The effects of two dihydropyridine calcium channel antagonists (nitrendipine and nifedipine) and two agonists (Bay K8644 and CGP-28392) were tested on high K(+)-induced contractures of frog's toe muscles. All four drugs depressed or blocked maximum contractures induced by 123 mM K+. Agonist effects, i.e., an increase in contracture amplitude, were found with smaller contractures produced by lower high K+ concentrations (i.e., 10, 20, and 25 mM). Bay K8644 produced its maximum agonist effect at 10(-7) M and only depressed contractures with 10(-6) M. CGP-28392 had its greatest agonist effect with 10(-9) M and had only antagonist effects with 10(-7) M or more. Nitrendipine had no agonist effects but nifedipine produced agonist effects with all concentrations tested (10(-9) to 10(-4) M). These results support previous results indicating that these contractures are initiated by extracellular Ca2+ ions entering via the voltage-sensitive, slow calcium channels in the t-tubules. The results obtained in the present study also are consistent with the known pharmacological effects of these drugs on calcium channels.

Mechanisms of Action of Endothelin on Isolated Feline Cerebral Arteries: In vitro Pharmacology and Electrophysiology

Vascular endothelium has been found to produce a strong and potent vasoconstrictor peptide, endothelin. In this study, we have examined basic mechanisms underlying the contractile response of cerebral vessels to endothelin using in vitro pharmacology and electrophysiology. It was found that endothelin produced strong concentration-dependent contractions of circular segments of the feline middle cerebral artery. The response was slow in onset and long lasting. The vessels showed a remarkably strong tachyphylactic reaction upon repeated exposure to endothelin. The contractile effect of endothelin was not modified by the alpha-adrenoceptor antagonist phentolamine (10(-6) M) or the 5-hydroxytryptamine antagonist ketanserin (10(-6) M). Mechanical removal of the endothelium decreased potassium contractions while the maximum response to endothelin was only slightly reduced. There was no change in sensitivity of the cerebral artery to endothelin. The addition of a calcium antagonist (10(-6) M diltiazem or 3 X 10(-8) M nimodipine) or removal of extracellular calcium from the buffer solution did not change the sensitivity of the artery to endothelin but the maximum response to endothelin was reduced by between 40 and 60% by these procedures. The resting membrane potential of the cat middle cerebral artery was -62.8 +/- 3.5 mV. There was no significant depolarization in conjunction with cumulative administration of endothelin in concentrations below 1 X 10(-9) M. However, bursts of excitatory junction potentials were occasionally seen in response to high concentrations of endothelin (5 X 10(-9) M).(ABSTRACT TRUNCATED AT 250 WORDS)

Direct effects of graded hypoxia on intact and denuded rabbit cranial arteries.

In isolated rabbit common carotid, internal carotid, and basilar arteries denuded of endothelium, mounted for measurement of contractile activity, and contracted with 10 microM serotonin (common and internal carotid) or 100 microM uridine 5'-triphosphate (UTP; basilar), 20 min of severe (PO2 = 15 Torr) and moderate (PO2 = 35 Torr) hypoxia relaxed initial tensions to 17, 6, and 16% and 18, 7, and 61% of control, respectively. The corresponding values in arteries contracted with 120 mM potassium-Krebs solution were 42, 60, and 73% and 57, 72, and 88%. These data indicate that the main determinants of the responses to hypoxia were intrinsic to the vessel walls of these arteries and that complete depolarization attenuated but did not eliminate the effects of these mechanisms. Superimposed on these intrinsic mechanisms were the effects of the endothelium, which were evaluated by several methods, including integration of the area between the 20-min response time courses of corresponding intact and denuded segments. Positive and negative integrated area values indicated endothelial vasoconstrictor [endothelium-derived contracting factor (EDCF)] and vasodilator [endothelium-derived relaxing factor (EDRF)] influences, respectively. In common carotid, internal carotid, and basilar segments contracted with serotonin and/or UTP, these areas averaged 770, 354, and 44 min% during severe and 491, 189, and -411 min% during moderate hypoxia. Corresponding values during potassium contraction were 217, -271, and -356 min% and 52, -177, and -54 min%. Together, these findings suggest that 1) intrinsic vascular mechanisms contribute significantly to hypoxic cerebral vasodilation in the rabbit, 2) EDCF is more prominent but also may be more sensitive to depolarization than EDRF, 3) hypoxia promotes the simultaneous release of both EDCF and EDRF, and 4) the ratio of EDCF to EDRF released during hypoxia decreases as one moves from the rabbit common carotid to the internal carotid to the basilar arteries.

Effects of cobalt, magnesium, and cadmium on contraction of rat soleus muscle

The effects on isometric tension of three divalent ions that block calcium channels, magnesium, cobalt, and cadmium, were tested in small bundles of rat soleus fibers. Cobalt, at a concentration of 2 or 6 mM, reversibly depressed twitch and tetanic tension and the depression was much greater in solutions containing no added calcium ions. Magnesium caused much less depression of tension than cobalt. The depression of tension was not accompanied by membrane depolarization or a reduction in the amplitude of action potentials. A reduction caused by 6 mM cobalt in the amplitude of 40 or 80 mM potassium contractures was not accompanied by a comparable reduction in tension during 200 mM potassium contractures, and could be explained by a shift in the potassium contracture tension-voltage curve to more positive potentials (by +7 mV on average). Similar effects were not seen with 2 or 6 mM magnesium. At a concentration of 20 mM, both cobalt and magnesium depressed twitch and tetanic tension, cobalt having greater effect than magnesium. Both ions shifted the potassium contracture tension-voltage curve to the right by +5 to +10 mV, caused a small depression of maximum tension, and slowed the time course of potassium contractures. Cadmium (3 mM) depressed twitch, tetanic, and potassium contracture tension by more than 6 mM cobalt, but experiments were complicated by the gradual appearance of large contractures that became even larger, and sometimes oscillatory, when the solution containing cadmium was washed out. It was concluded that divalent cations affect both activation and inactivation of tension in a manner that cannot be completely explained by a change in surface charge.

Noninactivating tension in rat skeletal muscle. Effects of thyroid hormone.

Inactivation of excitation-contraction coupling was examined in extensor digitorum longus (EDL) and soleus muscle fibers from rats injected daily with tri-iodothyronine (T3, 150 micrograms/kg) for 10-14 d. Steady-state activation and inactivation curves for contraction were obtained from measurements of peak potassium contracture tension at different surface membrane potentials. The experiments tested the hypothesis that noninactivating tension is a "window" tension caused by the overlap of the activation and inactivation curves. Changes in the amplitude and voltage dependence of noninactivating tension should be predicted by the changes in the activation and inactivation curves, if noninactivating tension arises from their overlap. After T3 treatment, the area of overlap increased in EDL fibers and decreased in soleus fibers and the overlap region was shifted to more negative potentials in both muscles. Noninactivating tension also appeared at more negative membrane potentials after T3 treatment in both EDL and soleus fibers. The effects of T3 treatment were confirmed with a two microelectrode voltage-clamp technique: at the resting membrane potential (-80 mV) contraction in response to a brief test pulse required less than normal depolarization in EDL, but more than normal depolarization in soleus fibers. After T3 treatment, the increase in contraction threshold at depolarized holding potentials (attributed to inactivation) occurred at more depolarized holding potentials in EDL, or less depolarized holding potentials in soleus. The changes in contraction threshold could be accounted for by the effects of T3 on the activation and inactivation curves. In conclusion, (a) T3 appeared to affect the expression of both activation and inactivation characteristics, but the activation effects could not be cleanly distinguished from T3 effects on the sarcoplasmic reticulum and contractile proteins, and (b) the experiments provided evidence for the hypothesis that the noninactivating tension is a steady-state "window" tension.

Na/Ca exchange and excitation--contraction coupling in frog fast fibres.

A 3 Na/Ca exchanger in the transverse tubular wall is modelled as the coupling mechanism between transverse tubular depolarization and Ca release from the sarcoplasmic reticulum. At rest, the Ca-occupied site faces the transverse tubular lumen. Upon depolarization, the difference in chemical potentials of Na and Ca gives a net inward force on Ca resulting in a reorientation of the exchanger so the Ca site now faces the myoplasm and releases Ca to stimulate Ca-induced Ca release from the sarcoplasmic reticulum. The rotation of the exchanger's asymmetrical charge could generate the 'charge movement' signal. As depolarization continues, the site is depleted of Ca and contraction ends spontaneously. Repolarization reorients the exchanger; the depleted Ca site now faces the transverse tubular lumen and slowly refills with Ca (repriming). A kinetic model is capable of controlling both twitch and contracture tension. The Na/Ca exchange blocker dichlorobenzamil (Merck) (10 microM), elevated external Na and low pH all slowed the rate of rise of potassium contracture tension. The ratios of rates of tension rise were dCB/control = 0.4 +/- 0.1, elevated external Na/Tris = 0.6 +/- 0.1, pH 6.3/control = 0.7 +/- 0.01. These results can be mimicked with the kinetic model by slowing the rate of 'rotation' (and hence charge movement) by 50%. Elevated internal Na increases the rate of rise of contracture tension; elevated internal Na/control 1.6 +/- 0.3. Dichlorobenzamil also slows the recovery following spontaneous relaxation; the time constant (68 s) of repriming is unchanged but shifted to longer recovery times. Reduced external Na and pH 6.3 also slow recovery in a similar manner, consistent with delayed rotation of the Ca-depleted site. These results suggest that Na/Ca exchange is a step in both the excitation contraction coupling chain and the repolarization-repriming sequence.

Nitrendipine blocks high potassium contractures but not twitches in rat skeletal muscle.

The effects of the organic calcium channel blocker nitrendipine was tested on electrically evoked twitches and on potassium depolarization-induced contractures of rat lumbricalis muscles. Nitrendipine (10(-7) to 5 X 10(-5) M) blocked only the potassium contractures. It was concluded that blocking calcium uptake through the slow voltage-sensitive calcium channels during potassium depolarization blocks the mechanical response of the muscle. Thus extracellular calcium ions are required for the excitation-contraction (E-C) coupling during depolarization contractures. On the other hand, electrically evoked twitches were not affected by nitrendipine; therefore, extracellular calcium ions entering via the slow voltage-sensitive channels are not required for E-C coupling during the twitch.

The effect of D600 on potassium contractures of slow muscle fibres of Rana temporaria.

(1) The effect of 30 microM D600 on the amplitude and time course of isometric contractures was studied in single slow fibres of Rana temporaria. (2) D600 only slightly reduced the amplitude of contractures evoked with 30 or 95 mM K-Ringer's. Maintenance of tension was strongly impaired by D600 only during exposure to 95 mM K. The caffeine contracture was not affected. (3) Addition of 10 mM Ca2+ or other divalent cations to the medium strongly counteracted the effect of D600 on maintained tension. The order of efficiency was Ca2+ = Ni2+ greater than Co2+ greater than Mn2+ much greater than Mg2+. (4) During 2 min exposure to 95 mM K-Ringer's the slow fibres inactivated to a variable degree; recovery from inactivation in normal Ringer's proceeded with a half time of the order of 1 min, while in the presence of D600 recovery was prolonged 3.3 to 27 times. (5) It is concluded that the effect of D600 on the contractile behaviour of slow fibres from Rana temporaria is predominantly due to a prolongation of the inactivated state. It is suggested that D600 binds to a site at the outer membrane surface which also binds divalent cations and determines the degree of contractile inactivation during exposure to potassium. Blocking of Ca2+ channels is unlikely to be the mechanism of this D600-effect.

The effect of diazepam on potassium contractures, contraction threshold, and resting tension in rat skeletal muscles.

The effects of diazepam on potassium contractures, contraction threshold, and resting tension have been examined in rat soleus muscle fibres. Two actions of the drug were defined that could not be attributed to changes in the resting membrane potential or depolarization in high potassium solutions. The major effect was an increase in the amplitude of submaximal tension during either twitches or potassium contractures and an increase in resting tension. At 400 microM diazepam, there was (a) a fourfold increase in 40 mM potassium contracture tension, (b) a negative shift of 8 mV in the membrane potential for half maximum tension estimated from the best fit of a Boltzmann-type equation to average potassium contracture data, (c) a negative shift of 8 mV in the threshold for contraction measured under voltage clamp conditions, and (d) a contracture of variable amplitude to a level that was occasionally equivalent to maximum tetanic tension. These potentiating actions of diazepam depended on drug concentration within the range of 100-800 microM. In contrast, the second effect of diazepam, depression of maximum tension by 10-15%, was independent of drug concentration between 100 and 400 microM. The results support the idea that diazepam produces an increase in resting myoplasmic calcium concentrations.

Effects of extracellular calcium concentration and dihydropyridines on contraction in mammalian skeletal muscle.

1. Twitches, tetanic contractions and potassium contractures were recorded isometrically from small bundles of rat soleus muscle fibres. 2. Solutions with reduced calcium concentrations (low-calcium solutions), whether buffered with EGTA (85 and 3 microM-Ca2+) or not (15 microM-Ca2+), caused an initial potentiation of contraction followed by depression. 3. The decay of potassium contractures (200 mM-potassium) was more rapid than normal in low-calcium solutions. 4. Recovery from the inactivation produced by a 200 mM-potassium contracture was slowed in low-calcium solutions but full recovery was seen within 10-15 min after return to a solution containing 2.5 mM-Ca2+. 5. Nifedipine (50 microM) in solutions containing 2.5 mM-Ca2+ potentiated contraction whereas, in low-calcium solutions, contraction was depressed and the depression was more pronounced the lower the Ca2+ concentration. 6. As with low-calcium solutions, potassium contractures decayed more rapidly in solutions containing nifedipine. Nifedipine slowed still further the rate of recovery from inactivation in low-calcium solutions. 7. (-) Bay K 8644 (50 microM) depressed contraction, increased the rate of decay of potassium contractures and slowed recovery from inactivation, like nifedipine. The racemate of Bay K 8644 was less effective. 8. In explanation of these and other observations, it is proposed that there is a dihydropyridine-binding molecule in the walls of the transverse tubular system that normally exists predominantly in a 'precursor' form at the resting membrane potential and is converted by membrane depolarization to an 'activator' form essential for excitation-contraction coupling. Conversion of the precursor to activator involves both conformational change and dissociation of calcium. Prolonged depolarization converts activator to an inactivated form by inducing further conformational change and dissociation of calcium. Recovery from inactivation requires reverse conformational changes and rebinding of calcium. The dihydropyridines affect contraction by reducing the affinity of the molecule for calcium.

Heat production of quiescent ventricular trabeculae isolated from guinea-pig heart.

1. A new calorimetric technique has been developed which allows continuous measurement of the rate of energy expenditure in superfused preparations of cardiac muscle. Thin trabeculae of guinea-pig ventricular muscle were mounted in a Perspex tube of 0.8 mm inner diameter and the temperature difference of the perfusate upstream and downstream of the preparation was measured. 2. The resting heat rate of trabeculae of 240-575 microns diameter from guinea-pig heart was determined repeatedly for up to 6 h after cardiectomy. It did not vary with time during the course of the experiment. 3. The average resting heat rate measured in HEPES-buffered Tyrode solution containing 20 mM-glucose and 2 mM-pyruvate as substrates was 130 +/- 29 mW/g dry weight or 36 +/- 8 mW/cm3 of tissue (n = 15). This is an order of magnitude larger than the resting heat rate reported in the literature for isolated cardiac preparations. 4. After omitting the pyruvate from the superfusate the resting heat rate decreased to 60-70% of its steady value within 4 min. After readmission of pyruvate this effect was reversed. The average resting heat rate with glucose as sole substrate was 23 +/- 4 mW/cm3. 5. Uncoupling of the mitochondria by 50 microM-2,4-dinitrophenol (DNP) increased the heat rate up to 170 mW/cm3. This effect could be maintained for several minutes and was fully reversible. Raising the external K+ concentration to 150 mM (NaCl replaced by KCl) induced a transient rise in the rate of heat production up to 115 mW/cm3. 6. The heat production during uncoupling of the mitochondria and during potassium contractures was inversely related to the diameter of the preparation. Calculation based on Hill's equation (Hill, 1928) indicated that this was caused by the development of anoxia at the core of the preparation. 7. In contrast, the rate of heat production of quiescent preparations was not correlated with diameter and calculation indicated that at rest there was no anoxic core. The high value of resting heat rate found in the present study is discussed within the context of the large variation of 1.7-25 mW/g reported in the literature for resting metabolic rate of cardiac muscle.

The effect of lithium and nitrate on potassium contractures in a glycerol-treated slow-twitch muscle of the rat.

Glycerol treatment (0.4 or 1.2 M) altered but did not abolish twitch, tetanus, potassium, or caffeine contractures in the rat soleus at 37 degrees C. The use of nitrate Krebs or lithium Krebs greatly reduced the effects of 1.2 M glycerol treatment on potassium contractures. The data indicate that the rat soleus is relatively resistant to the uncoupling effects of glycerol treatment and that both lithium and nitrate largely prevent these effects.

Potassium contractures in the mouse diaphragm: regional variation in the multiphasic response.

1. Submaximal potassium-induced contractures were studied in isolated mouse hemidiaphragms (HD) and compared with extensor digitorum longus (EDL) and soleus. 2. In the HD, the contracture was triphasic: an initial small, fast phase followed by a larger slow phase, and a late phase which was seen on prolonged exposure. 3. Costal segments of the HD responded to elevated potassium with a larger fast phase, while dorsal or ventral segments produced a larger slow phase. 4. In calcium-free solution, the fast phase appeared to be unaffected, while the slow phase was potentiated and its time course abbreviated. The late phase was greatly attenuated or abolished. 5. In the soleus, but not the EDL muscle, contracture amplitude in calcium-free solution was potentiated and the time course was shortened. 6. The characteristics of potassium-induced contractures of both the soleus and EDL muscles are to be seen in the response of the hemidiaphragm, but there is a regional variation in their proportional contribution which suggests a heterogeneity in diaphragm composition.

Paralysis of skeletal muscle by butanedione monoxime, a chemical phosphatase.

The chemical phosphatase butanedione monoxime (BDM) reversibly inhibited twitches and tetanic contractions in bundles of rat soleus fibres in a dose-dependent manner (2-20 mM) but had no effect on the amplitude or time course of action potentials. In addition, BDM reversibly reduced the amplitude of potassium contractures demonstrating a depressant effect on contraction not mediated by action potentials. BDM had no effect on asymmetric charge movement but depressed calcium currents across the surface membrane in voltage-clamped fibres. The most significant effect of BDM on excitation-contraction coupling was a reduction in the amplitude of the calcium transient associated with contraction in aequorin-injected fibres. While these experiments do not eliminate the possibility of a direct effect of BDM on contractile filaments, reduction of calcium release from the sarcoplasmic reticulum, at least at low concentrations of BDM (below 2 mM), would seem to be the main mechanism for the inhibition of contractions in rat skeletal muscle.

Contraction of rat skeletal muscle after glycerol treatments.

The effects of glycerol (400 to 1,200 mM) treatment on contractions of rat soleus muscles were investigated. Glycerol induced temperature- and concentration-dependent contractures which depended largely on extracellular calcium. Glycerol treatments reduced but did not abolish twitch, tetanus, and potassium contracture tension and these changes were far less at 22 than at 37 degrees C. Caffeine contractures were not altered following glycerol treatment at 22 degrees C, but were at 37 degrees C. It is suggested that the lesser effects of glycerol treatment on the soleus compared to other muscles may be due to its smaller transverse tubule system. Glycerol permeation and thus its osmotic action may be less in the soleus than in other muscles.

Effect of divalent cations on the potassium contracture of slow muscle fibers ofRana temporaria

K contractures of single slow muscle fibers ofRana temporaria were measured isometrically in the presence of normal, reduced, and increased Ca2+ concentrations at 18 to 20°C. At normal Ca2+ concentration (1.8mm) contracture tension decreased from its peak value of 35.4±8.2 N/cm2 to 59.4±23.9% within one minute, and to 48.3±27% within two minutes (30 fibers). Peak tension was virtually unaffected by changes of the Ca2+ concentration, but maintenance of tension was impaired by low (0.2mm), and improved by high (10mm) Ca2+ concentrations. When Ca2+ was added during the K contracture, there was practically no restoration of lost tension. Effects similar to those of Ca2+ were observed upon addition of foreign divalent cations to the medium. Co2+, Ni2+, and Cd2+ were slightly more effective than Ca2+ and Mn2+; the smallest effects were obtained with Mg2+, Sr2+, and Ba2+. The effects of foreign divalent cations were independent of the presence of Ca2+. It is concluded that in slow fibers ofRana temporaria maintenance of contracture tension is not due to an influx of Ca2+ ions. Instead, binding of divalent cations to superficial sites seems to be essential.

Potassium contractures and asymmetric charge movement in extensor digitorum longus and soleus muscles from thyrotoxic rats.

Potassium contractures and asymmetric charge movement were recorded from extensor digitorum longus (EDL) and soleus muscle from normal rats and rats that had been made thyrotoxic by daily intraperitoneal injections of triiodothyronine (150 micrograms kg-1) for two to three weeks. Potassium contracture tension (relative to tetanic tension) in thyrotoxic rats was greater in EDL muscles and smaller in soleus muscles than in normal rats. As the relationship between membrane potential and potassium concentration was unaltered by thyroid treatment, it was concluded that the changes in potassium contracture tension were due to changes in excitation-contraction coupling. In thyrotoxic rats there was an average negative shift of -5 mV in the voltage sensitivity of tension in EDL fibres and a positive shift of 5 mV in soleus. As a result, the tension-membrane potential curves for EDL and soleus fibres essentially coincided. There was a corresponding average negative shift of -4 mV in the voltage sensitivity of asymmetric charge in EDL fibres, and a positive shift of 4 mV in soleus fibres from thyrotoxic rats. The dependence of asymmetric charge movement on membrane potential became essentially the same in EDL and soleus fibres from thyrotoxic rats. The maximum asymmetric charge in soleus fibres increased from an average value of 6.5 nC microF-1 in normal rats (n = 33) to 8.9 nC microF-1 in thyrotoxic rats (n = 32; p less than 0.005).(ABSTRACT TRUNCATED AT 250 WORDS)

Suppression of charge movement in frog skeletal muscle by D600

Charge movements in intact frog twitch fibres were studied using a three-microelectrode voltage-clamp technique. When high potassium solution was applied transiently to the muscle fibres at low temperature in the presence of D600, the fibres became paralysed and, concomitantly, charge movement disappeared. The amount of charge suppressed by the paralysis treatment was about 70-100% of that in control experiments. This paralysing action of D600 is not shared by its derivative D890. The requirement of conditioning potassium contracture is, most likely, related to prolonged membrane depolarization, as voltage-clamped depolarization to 0 mV lasting tens of seconds also suppressed charge movement. When paralysed fibres were warmed, the main charge component (Q beta) was reprimed. By contrast, the hump charge component (Q gamma) was only reprimed in some of the fibres. Other than by warming, as paralysed fibre could be revived by stimulating it with large suprathreshold pulses but not by voltage-clamped hyperpolarization to -160 mV for tens of seconds. The paralysing action of D600 described here appears to be unrelated to its ability in blocking Ca2+ channels.

Inhibition of excitation-contraction coupling by a Ca channel blocker nicardipine at low temperature in frog twitch fibers.

Effects of a Ca channel blocker, nicardipine, on excitation-contraction (E-C) coupling were investigated in single twitch fibers dissected from short skeletal muscles of the frog, Rana japonica. The treatment with 20 microM nicardipine at 25 degrees C evoked a reversible twitch potentiation. By contrast, the treatment at the same concentration at 5 degrees C for 30 min evoked an intense twitch inhibition. The inhibition was irreversible. This paralyzed fiber remained at nearly normal resting and propagated action potentials. The treatment with 30 microM nicardipine for 14 min at 6 degrees C completely inhibited a potassium contracture by 190 mEq/l potassium ion for 1.5 h after the removal of nicardipine despite the retention of normal resting potentials. The paralyzed fiber which was previously treated with 20 microM nicardipine for 30 min at 6 degrees C, also remained at normal action potential and at partial potassium contracture, and responded to 3 mM caffeine stimulation with a normal contracture. Dose-dependence curves obtained from the data on the treatments with nicardipine at various concentrations for 30 min at 6 degrees C consisted of an opposite sigmoidal shape with both threshold and half maximum inhibition at 1 and 1.5 microM for twitch and similarly at 5 and 12 microM for potassium contracture. Temperature-dependence curves obtained from the data on the treatments with 20 microM nicardipine for 30 min at various temperatures consisted of a sigmoidal shape with half maximum inhibition at 15 degrees C for twitch and at 12 degrees C for potassium contracture. A working hypothesis "two component-three state model" for E-C coupling was proposed based on the interpreted mechanism of nicardipine action.