Biphasic potassium contractures in frog muscle fibers

Potassium-induced contractures were studied in single fibers from the semitendinosus muscle of Rana pipiens. Contractures elicited by solutions containing 60–117 mM potassium and 120 mM chloride were biphasic, consisting of a rapid initial contraction with a duration at 23°C of less than 1 sec followed by a slow response with a duration of many seconds. At 13°C, the initial response was greatly prolonged so that the two responses virtually fused into a single smooth contracture. Membrane potential in high potassium, high chloride solutions underwent a transient peak depolarization, probably as a result of time-dependent changes in membrane conductance during depolarization. It is proposed that this complex time course of depolarization gives rise to the biphasic contracture response.     

Determination of ionic calcium in frog skeletal muscle fibers

Ionic calcium concentrations were measured in frog skeletal muscle fibers using Ca-selective microelectrodes. In fibers with resting membrane potentials more negative than -85 mV, the mean pCa value was 6.94 (0.12 microM). In fibers depolarized to -73 mV with 10-mM K the mean pCa was 6.43 (0.37 microM). This increase in the intracellular [Ca2+] could be related to the higher oxygen consumption and heat production (Solandt effect) reported to occur under these conditions. Caffeine, 3 mM, also produced an increase in the free ionic calcium to a pCa of 6.52 (0.31 microM) without changes in the membrane potential. Lower caffeine concentrations, 1 and 2 mM, did not change the fiber pCa. Lower Ca concentrations in the external medium effectively reduced the internal ionic calcium to an estimated pCa of 7.43 (0.03 microM).     

Stimulation frequency and external ionic composition control the repriming of caffeine-induced contractures in frog skeletal muscle

The effect of stimulation rate and of external ionic composition on the repriming period of contractures induced by 6 mM caffeine was tested on isolated skeletal muscle fibres of the frog (Rana ridibunda). The repriming period, which was 11.2 +/- 0.1 min (mean +/- SEM, n = 9) on quiescent fibres, was shortened in fibres stimulated at a frequency ranging from 3 to 12 min-1 (optimal rate, 8 min-1; full repriming 5.7 +/- 0.2 min; n = 10). A 10-fold increase in the extracellular calcium concentration shortened the repriming period on both stimulated and quiescent fibres, whereas decreasing external calcium (1/10) delayed it. In a Na+-free solution (Li+ substituted) the repriming period of stimulated fibres was markedly delayed (14 min), whereas quiescent fibres never recover more than 10% of their ability to develop subsequent caffeine contractures. In contrast, with a 35% Na solution, the repriming period was greatly shortened (stimulated, 5.4 +/- 0.2 min, n = 7; quiescent, 6.2 +/- 0.5 min, n = 8). It is concluded that repriming depends on three mechanisms that seem to refill a calcium store and trigger recovery: the slow inward calcium current, a Na+-Ca2+ exchange, and perhaps a passive Ca2 influx.     

Chloride and osmotic contractures in skinned frog muscle fibers

Summary Single contractures were elicited in segments of skinned frog muscle fibers when the segments were moved from relaxing-loading solutions to various test solutions. The effective test solutions produced an increase in the concentration of chloride ions in the myofilament space, [Cl] _ms , and/or presumably caused the sarcoplasmic reticulum to undergo a change in volume. The contractures were quantified in terms of their maximum tension and time-integral. Two outer segments from each fiber underwent a contracture in a control solution (chloride ions were substituted for all of the methanesulfonate ions in the relaxing solution). The mean values of tension and area in the control contractures of each fiber were divided into the corresponding values from a test contracture obtained in the central segment of the same fiber. Test contractures obtained upon increasing [Cl] _ms and increasing the product, [K] _ms ×[Cl] _ms , were compared to contractures that were obtained by increasing [Cl] _ms while keeping [K] _ms ×[Cl] _ms constant. The former contractures were greater in magnitude for a given [Cl] _ms . Whereas the former solutions may have caused an increase in the volume of the sarcoplasmic reticulum and altered the electrical potential across the membranes of the sarcoplasmic reticulum as well, only a change in potential was presumed to have occurred in the latter solutions. Other types of contractures were investigated to show that both swelling of the sarcoplasmic reticulum and changes in the electrical potential of its membranes can cause release of calcium ions and elicit contractures in skinned fibers.     

Effects of agonists and antagonists of rhyanodine receptors on potassium contractures in twitch and tonic frog skeletal muscle fibers

A comparative analysis of the effects of the concentrations of Ca2+ in external medium and the inhibitor (dantrolene) and activator (4-chloro-m-cresol) of rhyanodine-sensitive Ca2+ channels of carcoplasmic reticulum on the characteristics of potassium contracture in frog twitch and tonic skeletal muscles has been performed. It was shown that the duration of contracture in tonic muscles is not restricted by the presence of Ca2+, as distinct from twitch muscles. Dandrolene does not practically affect the contractile responses of tonic fibres, and the concentration of cresol eliciting the contracture for tonic fibres is substantially higher (1 mM) than for twitch fibers (0.25 mM). In twitch fibers, the potassium contracture activated in the presence of cresol is comparable in amplitude and dynamics with the contracture under control conditions, and in tonic fibers a summing of responses without relaxation after the washing of excessive potassium is observed. This suggests that, in twitch fibers, the influx of Ca2+ can directly create the concentration sufficient for the maintenance of contraction, and in tonic fibers its involvement is mediated through the Ca(2+)-dependent activation of the beta-isoform of rhyanodine-sensitive channels.     

Effects of Na-octanoate on potassium contractures in normal and denervated frog tonic fibres

In frog twitch muscle fibres, Na-octanoate (NaC8) shifted the relation between potassium induced tension and membrane potential to the right. The present study has been carried out to investigate the effect of this fatty acid on frog tonic fibres. Potassium contractures measured on bundles of 30-40 fibres of ileofibularis muscles were less decreased by NaC8 (2.5-10 mmol/l) than those of twitch fibre bundles. In denervated muscles the sensitivity to NaC8 was increased, probably due to the development of sodium channels in the membranes. Experiments with mixed fibre bundles also showed a lower influence of NaC8 on potassium contracture of tonic fibres. On the other hand, tonic fibres showed a lower threshold of the potassium induced tension as well as a lower K+ concentration for maximal activation. This lower threshold was further lowered by NaC8, corresponding to a shift of the relation between potassium concentration and tension to the left. The membrane resting potentials were -58 +/- 9 mV in tonic fibres and -83 +/- 5 mV in twitch fibres. Five mmol/l NaC8 only induced depolarization of the membrane of tonic fibres. This depolarization (by about 20 mV) may be responsible for the threshold shift to lower K+ concentration in NaC8-exposed tonic fibres. In addition to the effects of NaC8 on sodium channels, interactions with Ca2+ binding sites are discussed.     

Effects of potassium,sodium, and azide on the ionic movements that accompany activity in frog nerves

Stimulation of intact or desheathed frog sciatic nerves produced an increase in the sodium content and a decrease in the potassium content of this tissue. In desheathed preparations the magnitudes of the changes in ionic contents decreased as the concentration of the potassium in the bathing solution was increased, while changing the external sodium concentration produced small effects on the ionic shifts. During tetanization, the rate of decline of the compound action potential also decreased as the external potassium concentration increased. Eliminating the activity respiration with 0.2 mM azide did not greatly modify the changes in sodium and potassium distribution that accompanied activity in either intact or desheathed nerves.     

Model of calcium movements during activation in the sarcomere of frog skeletal muscle

A model of calcium movement during activation of frog skeletal muscle is described. The model was based on the half sarcomere of a myofibril and included compartments representing the terminal cisternae, the longitudinal sarcoplasmic reticulum, the extramyofibrillar space, and the myofibrillar space. The calcium-binding proteins troponin, parvalbumin, and calsequestrin were present in appropriate locations and with realistic binding kinetics. During activation a time-dependent permeability in the terminal cisternal wall led to calcium release into the myoplasm and its diffusion through the myoplasm longitudinally and radially was computed. After adjustment of three parameters, the model produced a myoplasmic free-calcium concentration that was very similar to those recorded experimentally with calcium indicators. The model has been used to demonstrate the importance of parvalbumin in the relaxation of skeletal muscle, to describe the time course and magnitude of calcium gradients associated with diffusion across the sarcomere, and to estimate the errors associated with the use of aequorin as an intracellular calcium indicator in muscle.     

Contraction transients of skinned muscle fibers: effects of calcium and ionic strength

Calcium and ionic strength are both known to modify the force developed by skinned frog muscle fibers. To determine how these parameters affect the cross-bridge contraction mechanism, the isotonic velocity transients following step changes in load were studied in solutions in which calcium concentration and ionic strength were varied. Analysis of the motion showed that calcium has no effect on either the null time or the amplitude of the transients. In contrast, the transient amplitude was increased in high ionic strength and was suppressed in low ionic strength. These results are consistent with the idea that calcium affects force in skeletal muscle by modulating the number of force generators in a simple switchlike "on-off" manner and that the steady force at a given calcium level is proportional to cross-bridge number. On the other hand, the effect of ionic strength on force is associated with changes in the kinetic properties of the cross-bridge mechanism.     

The temperature dependence of ionic influences on contractile function in frog twitch muscle

Alterations in the ionic composition of the medium produce striking changes in the potential-dependent contractile responses of skeletal muscles. This study was undertaken to examine the temperature dependence of some of these effects. The suppression of maximal K contractures of frog toe muscles in media lacking divalent cations was largely overcome by a sufficient increase in temperature. The restoration of K contractures by perchlorate in the absence of divalent cations was prevented by a sufficient decrease in temperature. The effect of perchlorate was to shift the temperature dependence of these contractures toward lower temperatures. The reduction in the amplitude of maximal K contractures in the absence of divalent cations was less marked after pretreatment with a reagent (trinitrobenzenesulfonate) that selectively modifies free amino groups, although the temperature dependence of these contractures was unchanged. The reduction in the amplitude of K contractures in the presence of an organic anion (hexanoate) was partially antagonized both by an increase in temperature or by a decrease in temperature, effects that resemble those observed in solutions in which the divalent cation concentration was reduced. In chloride solutions, the relation between [K]0 and K contractures was shifted toward lower [K]0 by an increase in temperature, whereas in perchlorate solutions increased temperature produced a shift in the opposite direction. The shift in this relation toward lower [K]0 at reduced temperature, and the accelerated time course of K contractures with an increase in temperature were similar in perchlorate and in chloride solutions. Thermodynamic analysis by Arrhenius plots indicated that the influence of divalent cations and perchlorate anions on K contractures may be the result of their effects on hydrational factors.     

The effect of dihydropyridines on the strontium and calcium content in frog muscle

2,6-Dimethyl-3,5-dimethoxycarbonyl-4-(O-difluromethoxyphenyl)-1,4- dihydropyridine (ryodipine) (2 x 10(-4) M) significantly blocks the resting Sr++ uptake in sartorius and iliofibularis muscles of Rana temporaria without changes in Ca++ and Mg++ content. Dihydropyridine CGP (2.10(-4) M) increased both [Sp++] and [Ca++] (mumol/g dr. w.) by about 20 and 13%, respectively, as compared with paired control muscles, whereas [Mg++] remained unaffected. The [Sr++] changes are interpreted as specific dihydropyridine-effect on the Ca(++)-channels in the muscle membrane.     

Effect of external sodium and calcium on calcium efflux in frog striated muscle

Summary The effect of media with different ionic composition on calcium efflux from the dorsal head of semitendinosus muscles ofRana pipiens was studied. The reduction in the fractional loss of⁴⁵Ca, when going from normal Ringer's solution to an ONa–OCa medium, was 60%. Withdrawal of only Na or Ca from the external medium also caused a significant drop in the fractional loss (33% and 34%, respectively). The effect of different concentrations of Ca (studied in the absence of the external Na) was also studied. It was found that a linear function could describe the relationship between the calcium-dependent calcium efflux and the external calcium concentration. These results indicate that calcium efflux from frog muscle fibers consists of three major components: one that is dependent on the presence of calcium in the external medium, one that is dependent on the presence of sodium in the external medium, and one that persists in the absence of these two cations.