Feedback in the Contractile Mechanism of the Frog Heart

Shortening causes a transient decrease, extension an increase, in activity during contractures of the frog ventricle induced by high Ca or by isosmotic K solution. This is shown by the fact that, after the immediate passive shortening, the muscle is extended under isotonic conditions when the load is diminished, and that under isometric conditions quick release causes first a rapid drop, then a further, much slower, fall of tension. Increasing the load or stretching induce the opposite effects. At low temperatures all rapid changes in length produce oscillations of low frequency. These responses are due to a sensitive feedback mechanism similar to that previously demonstrated for insect fibrillar muscle. That this mechanism comes into play in the heart under normal conditions and controls the time-course of the twitch is demonstrated by the observation that relaxation begins earlier the greater the shortening. Thus, during afterloaded isotonic twitches the onset of relaxation is advanced as the load is diminished.     

A phenomenological model for estimating metabolic energy consumption in muscle contraction

A phenomenological model for muscle energy consumption was developed and used in conjunction with a simple Hill-type model for muscle contraction. The model was used to address two questions. First, can an empirical model of muscle energetics accurately represent the total energetic behavior of frog muscle in isometric, isotonic, and isokinetic contractions? And second, how does such a model perform in a large-scale, multiple-muscle model of human walking? Four simulations were conducted with frog sartorius muscle under full excitation: an isometric contraction, a set of isotonic contractions with the muscle shortening a constant distance under various applied loads, a set of isotonic contractions with the muscle shortening over various distances under a constant load, and an isokinetic contraction in lengthening. The model calculations were evaluated against results of similar thermal in vitro experiments performed on frog sartorius muscle. The energetics model was then incorporated into a large-scale, multiple-muscle model of the human body for the purpose of predicting energy consumption during normal walking. The total energy estimated by the model accurately reflected the observed experimental behavior of frog muscle for an isometric contraction. The model also accurately reproduced the experimental behavior of frog muscle heat production under isotonic shortening and isokinetic lengthening conditions. The estimated rate of metabolic energy consumption for walking was 29% higher than the value typically obtained from gait measurements.     

Mechanical control of the rising phase of contraction of frog skeletal and cardiac muscle

The effect of shortening on contractile activity was studied in experiments in which shortening during the rising phase of an isotonic contraction was suddenly stopped. At the same muscle length and the same time after stimulation the rise in tension was much faster, if preceded by shortening, than during an isometric contraction, demonstrating an increase in contractile activity. In this experiment the rate of tension rise determined in various phases of contraction was proportional to the rate of isotonic shortening at the same time after stimulation. Therefore, the time course of the isotonic rising phase could be derived from the tension rise after shortening. The rate of isotonic shortening was found to be unrelated to the tension generated at various lengths and to correspond closely to the activation process induced by shortening. The length response explains differences between isotonic and isometric contractions with regard to energy release (Fenn effect) and time relations. These results extend previous work which showed that shortening during later phases of a twitch prolongs, while lengthening abbreviates contraction. Thus the length responses, which have been called shortening activation and lengthening deactivation, control activity throughout an isotonic twitch.     

Isometric contractile properties and instantaneous stiffness of amphibian skeletal muscle in the temperature range from 0 to 20 degrees C

The isometric contractile properties of frog (Rana pipiens) and toad (Bufo bufo) sartorii have been studied over the temperature range from 0 to 20 degrees C. The isometric twitch tension was found to vary considerably between these two species and between muscles in the same species. Between 0 and 4 degrees C there was very little change in maximum isometric twitch tension. Between 4 and 12 degrees C several muscles from frog or toad showed a potentiation of twitch tension whereas others showed a decline. Over this temperature range the toad sartorii consistently demonstrated a greater potentiation. By 12 degrees C a steady decline in twitch tension in both muscles was seen as the temperature range the toad sartorii consistently demonstrated a greater potentiation. By 12 degrees C a steady decline in twitch tension in both muscles was seen as the temperature approached 20 degrees C. The maximum isometric tetanic tension recorded between 18 and 20 degrees C increased fractionally to an average of 1.504 +/- 0.029 (n = 4) for frog sartorii and to 1.377 +/- 0.008 (n = 5) for toad sartorii. The time to peak twitch tension and the half-relaxation time decreased markedly with an increase in temperature. Moreover, the half-relaxation time was reduced by a greater proportion than the time to peak twitch tension. Measurements of instantaneous stiffness by controlled velocity releases from the plateau of isometric tetani revealed that the large increase in isometric tetanus tension as the muscle was warmed was not accompanied by a corresponding increase in the total number of active cross-bridges. The possibility that a decreased availability of intracellular Ca2+ ions at the contractile sites contributing to the fall of isometric twitch tension at elevated temperatures is discussed. The possibility exists that at elevated temperatures a change inthe intrinsic contractile ability of the muscle occurs which produces an increased tension per cross-bridge.     

Mechanical control of the time-course of contraction of the frog heart

Changes in load during most phases of an isotonic contraction of the frog and turtle heart increased or decreased the duration of the twitch. It was abbreviated by a maintained increase or by a brief decrease in load. The relaxing effect of these procedures developed with a delay lasting more than a second under some conditions and will be called lengthening deactivation. The reverse procedures, a maintained diminution or a brief increase in load, increased the duration of the twitch. This effect will be called shortening activation. Although the termination of relaxation may be delayed or advanced by the mechanical interventions mentioned, the normal time- course of isotonic relaxation was always resumed later, regardless of the starting level of the load, making it possible to measure accurately changes in the duration of the twitch. The responses to changes in load produce positive feedback during the isotonic contraction and explain, at least in part, the difference in the time- course of isotonic and isometric contraction. The effects of changes in load were much smaller and briefer in the atrium than the ventricle.     

Contractile properties of the shortening rat diaphragm in vitro

Diaphragmatic fatigue has been defined in terms of the failure of the muscle to continue to generate a given level of tension. Appropriate shortening of the diaphragm is, however, just as important for adequate ventilation. In this study we have examined in vitro the contractile properties of the rat diaphragm under afterloaded isotonic conditions and the effect of fatigue on the ability of the diaphragm to shorten. Shortening of the muscle strips was found to depend on size of afterload, frequency of stimulation, duration of stimulation, and initial length of the muscle. The afterloaded isotonic length-tension relationship coincided with the relationship between length and active isometric tension only for relatively small afterloads. Fatigue of the muscle strips, induced by isometric or afterloaded isotonic contractions, was associated with a decline in the extent of shortening as well as a decrease in active isometric tension. Ability to shorten and ability to develop isometric tension did not decrease to the same extent under all conditions. We conclude that active shortening, as well as active isometric tension, is decreased by muscular fatigue and that changes in these properties can be different depending on experimental conditions. The results suggest that the definition of diaphragmatic fatigue should be expanded to include the ability of the muscle to shorten by an appropriate amount. The results also suggest that measurement of isometric performance may not provide a complete estimate of the overall performance of the fatigued diaphragm.     

Mechanical properties of muscles from Xenopus borealis following maintenance in organ culture

The mechanical properties of sartorii muscles from the toad Xenopus borealis were studied in organ cultures lasting up to 26 days. Isometric twitch and tetanic tensions diminished in force reaching half their initial values after 15 days in culture. In contrast, twitch time to peak time to half relaxation, twitch-tetanus ratio and muscle weight-body weight ratio, were unchanged. Maximum isotonic shortening velocity (Vmax) declined, with a half time similar to that of the isometric force. The fall in isometric tension is probably due to a breakdown of activation in some fibres. The change in Vmax could be due to a loss of functional sarcomeres in series with the tendons.     

The dynamic properties of mammalian skeletal muscle

The dynamic characteristics of the rat gracilis anticus muscle at 17.5°C have been determined by isotonic and isometric loading. For a fixed initial length these characteristics were represented either as a family of length-velocity phase trajectories at various isotonic afterloads or as a series of force-velocity curves at different lengths. An alternate method of viewing these data, the length-external load-velocity phase space, was also generated. When the muscle was allowed to shorten from different initial lengths, the velocity of shortening achieved at a given length was lower for longer initial lengths. The amount of departure was also dependent upon the isotonic load, the greater the load the greater the departure. The departures were not caused by changes in the elastic elements of the muscle or fatigue in the ordinary sense. When the behavior of the muscle was investigated at different frequencies of stimulation, the shortening velocity was a function of the number of stimulating pulses received by the muscle at a given frequency. The shortening velocity of the rat gracilis anticus muscle is, therefore, not only a function of load and length, but also of an additional variable related to the time elapsed from onset of stimulation.     

Active and passive components in the length-dependent stiffness of tracheal smooth muscle during isotonic shortening.

Contraction of smooth muscle tissue involves interactions between active and passive structures within the cells and in the extracellular matrix. This study focused on a defined mechanical behavior (shortening-dependent stiffness) of canine tracheal smooth muscle tissues to evaluate active and passive contributions to tissue behavior. Two approaches were used. In one, mechanical measurements were made over a range of temperatures to identify those functions whose temperature sensitivity (Q(10)) identified them as either active or passive. Isotonic shortening velocity and rate of isometric force development had high Q(10) values (2.54 and 2.13, respectively); isometric stiffness showed Q(10) values near unity. The shape of the curve relating stiffness to isotonic shortening lengths was unchanged by temperature. In the other approach, muscle contractility was reduced by applying a sudden shortening step during the rise of isometric tension. Control contractions began with the muscle at the stepped length so that properties were measured over comparable length ranges. Under isometric conditions, redeveloped isometric force was reduced, but the ratio between force and stiffness did not change. Under isotonic conditions beginning during force redevelopment at the stepped length, initial shortening velocity and the extent of shortening were reduced, whereas the rate of relaxation was increased. The shape of the curve relating stiffness to isotonic shortening lengths was unchanged, despite the step-induced changes in muscle contractility. Both sets of findings were analyzed in the context of a quasi-structural model describing the shortening-dependent stiffness of lightly loaded tracheal muscle strips.     

Heat, mechanics, and myosin ATPase in normal and hypertrophied heart muscle

In this paper we review our previous work on the myothermic economy of isometric force production in compensated cardiac hypertrophy secondary to pulmonary artery constriction (pressure overload) and/or thyrotoxicosis (volume overload). Hypertrophy-induced changes in isotonic and isometric twitch mechanics are correlated with accompanying changes in actin-activated myosin ATPase and heat liberation. Heat measurements were made with rapid, high-sensitivity thermopiles on right ventricular papillary muscles from normal and hypertrophied rabbit hearts. Total activity-related heat was separated into initial and recovery heat. Initial heat was separated into a tension-dependent component (TDH) relating to cross-bridge activity, and a tension-independent component (TIH) relating to excitation-contraction coupling. There were oppositely directed changes in most parameters studied in pressure overload hypertrophy (P) as compared with thyrotoxic hypertrophy (T). Thus, in P there was depression (30-50% in the rate of isometric force production, mechanical Vmax, TDH and TDH rate, myosin ATPase, TIH, and prolongation in time-to-peak twitch tension, whereas in T all parameters were oppositely changed except for no change in TIH. Thyrotoxicosis following pressure overload reversed the P-induced changes in all parameters. There was a direct, linear relation between in vitro actin-activated myosin ATPase and in vivo TDH. However, TDH per unit twitch tension or tension-time integral varied inversely with ATPase, making force production more economical than normal in P muscles and less economical than normal in T muscles. These cellular changes beneficially equip P hearts for slow, high-pressure, economical pumping the T hearts for fast, high-volume, uneconomical pumping. The differences are similar to those between slow and fast skeletal muscle and between neonatal and adult skeletal muscle. The mechanism of these changes is discussed in terms of an enzyme kinetic scheme of chemomechanical coupling in actomyosin interaction.     

Double Sucrose-Gap Method Applied to Single Muscle Fiber of Xenopus laevis

Passive electrical properties (internal conductance, membrane conductance, low frequency capacity, and high frequency capacity obtained from the foot of the action potential) of normal and glycerol-treated muscle of Xenopus were determined with the intracellular microelectrode technique. The results show that the electrical properties of Xenopus muscle are essentially the same as those of frog muscle. Characteristics of the action potential of Xenopus muscle were also similar to those of frog muscle. Twitch tension of glycerol-treated muscle fibers of Xenopus recovered partially when left in normal Ringer for a long time (more than 6 h). Along with the twitch recovery, the membrane capacity increased. Single isolated muscle fibers of Xenopus were subjected to the double sucrose-gap technique. Action potentials under the sucrose gap were not very different from those obtained with the intracellular electrode, except for the sucrose-gap hyperpolarization and a slight tendency toward prolongation of the shape of action potential. Twitch contraction of the artificial node was recorded as a change of force from one end of the fiber under the sucrose gap. From the time-course of the recorded force and the sinusoidal stress-strain relationship at varying frequencies of the resting muscle fiber, the time-course of isotonic shortening of the node was recovered by using Fourier analysis. It was revealed that the recorded twitch force can approximately be regarded as isotonic shortening of the node.     

In vivo behaviour of human muscle architecture and mechanomyographic response using the interpolated twitch technique

This study investigated the origin of curvilinear change in the superimposed mechanomyogram (MMG) amplitude of the human medial gastrocnemius muscle (MG) with increasing contraction intensity. The superimposed twitch amplitude, the superimposed MMG amplitude and the extent of fascicle shortening were measured using ultrasonic images of electrical stimulation during isometric plantar flexions at levels 20%, 40%, 60%, 80%, and 100% of the maximal voluntary contraction (MVC). The superimposed twitch amplitude, the superimposed MMG amplitude and the extent of fascicle shortening decreased with increasing contraction intensity. The superimposed MMG amplitude and the extent of fascicle shortening showed a curvilinear decrease, while the superimposed twitch amplitude showed a linear decrease at levels up to 80% of the MVC. There was a linear relationship between the superimposed MMG amplitude and the extent of fascicle shortening at different contraction intensities. These results indicate that the superimposed MMG amplitude reflects changes in the extent of fascicle shortening at different contraction intensities better than the superimposed twitch amplitude. Our study suggests that the origin of the curvilinear decrease of superimposed MMG amplitude is associated with a curvilinear decrease of the extent of fascicle shortening with increasing contraction intensity in the human MG.     

Velocity transients and viscoelastic resistance to active shortening in cat papillary muscle.

When isotonic force steps were applied to activated papillary muscles, the velocity was almost never constant. Early rapid shortening associated with the step persisted for 2-7 ms after the step ends. The early rapid shortening is attributed to lightly damped series elastic recoil and velocity transients of the contractile elements. In most steps, the subsequent velocity declines progressively with shortening, and most of the decline in velocity can be accounted for by compression of a viscoelastic element in parallel with the contractile elements. To demonstrate this, the time course of isotonic velocity was compared with a model in which the force-velocity characteristics of the contractile element were assumed to be constant, and the decline in velocity was due to increasing compression of the viscoelastic element. This model predicted the observed results except that the predicted velocities rose progressively above the measured values for steps to light loads applied late in the twitch, and fell below the velocity trace for heavy loads applied early in the twitch. These deviations would occur if rapid shortening caused deactivation late in the twitch, and if activation were rising early in the twitch. A conditioning step applied to the muscle during the rise of force depressed both isometric force and maximum velocity measured at the peak of force; isometric force was more depressed with later conditioning steps than with earlier steps, while maximum velocity was depressed by about the same extent with both early and late steps. This difference between the effects on isometric force and maximum velocity are explained by a combination of deactivation and viscoelastic load.     

The Effect of Shortening on the Time-Course of Active State Decay

The active state describes the force developed in a muscle when the contractile elements are neither lengthening nor shortening. Recently it was suggested that perturbations used to measure the active state also alter the time-course of the active state. The present research was undertaken to assess quantitatively the effect of two such perturbations, isotonic shortening and quick release, on the active state in frog sartorius muscle. Methods were developed which allowed the determination of active state points following periods of controlled isotonic shortening or quick release early in the contraction cycle. All experiments were carried out within the plateau region of the length-tension curve. Both isotonic shortening and quick release altered the active state decay. The active state force decreased as the extent of shortening or release was increased. For each 0.1 mm of isotonic shortening there was a 2% decrease in active state force. Quick release produced a larger decrement. From this data we conclude that the time-course of active state can be measured only in relative terms because it is altered by the motion which takes place in the contractile machine while the active state is being measured. This finding helps to resolve paradoxes in the literature relating to the time-course of the active state, calculated and experimentally determined isometric tetanic myograms, and the heat of shortening.     

Attenuated fatigue in slow twitch skeletal muscle during isotonic exercise in rats with chronic heart failure

During isometric contractions, slow twitch soleus muscles (SOL) from rats with chronic heart failure (chf) are more fatigable than those of sham animals. However, a muscle normally shortens during activity and fatigue development is highly task dependent. Therefore, we examined the development of skeletal muscle fatigue during shortening (isotonic) contractions in chf and sham-operated rats. Six weeks following coronary artery ligation, infarcted animals were classified as failing (chf) if left ventricle end diastolic pressure was >15 mmHg. During isoflurane anaesthesia, SOL with intact blood supply was stimulated (1s on 1s off) at 30 Hz for 15 min and allowed to shorten isotonically against a constant afterload. Muscle temperature was maintained at 37°C. In resting muscle, maximum isometric force (F(max)) and the concentrations of ATP and CrP were not different in the two groups. During stimulation, F(max) and the concentrations declined in parallel sham and chf. Fatigue, which was evident as reduced shortening during stimulation, was also not different in the two groups. The isometric force decline was fitted to a bi-exponential decay equation. Both time constants increased transiently and returned to initial values after approximately 200 s of the fatigue protocol. This resulted in a transient rise in baseline tension between stimulations, although this effect which was less prominent in chf than sham. Myosin light chain 2s phosphorylation declined in both groups after 100 s of isotonic contractions, and remained at this level throughout 15 min of stimulation. In spite of higher energy demand during isotonic than isometric contractions, both shortening capacity and rate of isometric force decline were as well or better preserved in fatigued SOL from chf rats than in sham. This observation is in striking contrast to previous reports which have employed isometric contractions to induce fatigue.     

In vivo and in vitro correlation of trachealis muscle contraction in dogs.

Maximal trachealis muscle shortening in vivo was compared with that in vitro in seven anesthetized dogs. In addition, the effect of graded elastic loads on the muscle was evaluated in vitro. In vivo trachealis muscle shortening, as measured using sonomicrometry, revealed maximal active shortening to be 28.8 +/- 11.7% (SD) of initial length. Trachealis muscle preparations from the same animals were studied in vitro to evaluate isometric force generation, isotonic shortening, and the effect of applying linear elastic loads to the trachealis muscle during contraction from optimal length. Maximal isotonic shortening was 66.8 +/- 8.4% of optimal length in vitro. Increasing elastic loads decreased active shortening and velocity of shortening in vitro in a hyperbolic manner. The elastic load required to decrease in vitro shortening to the extent of the shortening observed in vivo was similar to the estimated load provided by the tracheal cartilage. We conclude that decreased active shortening in vivo is primarily due to the elastic afterload provided by cartilage.     

The effects of isotonic contractions on the rate of fatigue development and the resting membrane potential in the sartorius muscle of the frog, Rana pipiens.

The goal of this study was to characterize how isotonic contractions affect the rate of fatigue development. Muscle bundles dissected from frog sartorius muscles were stimulated with 100-ms long train of pulses (0.5 ms, 6 V, 140 Hz). To measure the effect of the isotonic contractions, isometric tetanus were elicited at regular time intervals during the stimulation to fatigue. In general, isotonic contractions caused a faster decrease in tetanic force than isometric contractions. The difference in tetanic force between an isotonic and isometric fatigue increased gradually over a 20-min period to 7.9 and 13.5% at 0.04 and 0.1 trains/s (TPS), respectively. At 0.2, 0.5, and 1.0 TPS, the decrease in tetanic force was also faster during an isotonic fatigue, which resulted in an initial difference in tetanic force between the two types of fatigue. The difference did not exceed 18.5% and did not persist throughout the stimulation period; i.e., the difference disappeared before the end of the fatigue stimulation. The half-relaxation time was prolonged during fatigue development, and the prolongation was greater during an isotonic fatigue, except at 0.04 TPS. The increases in the half-relaxation time at 0.2, 0.5, and 1.0 TPS were followed by a decrease, and the decreases were especially pronounced during an isotonic fatigue at 0.5 and 1.0 TPS. The results showed for the first time that isotonic contractions cause a faster rate of fatigue development in frog sartorius muscles, and this effect depends on the frequency of stimulation.     

Effect of stretch and release on equatorial X-ray diffraction during a twitch contraction of frog skeletal muscle.

Time-resolved intensity measurements of the x-ray equatorial reflections were made during twitch contractions of frog skeletal muscles, to which stretches or releases were applied at various times. A ramp stretch applied at the onset of a twitch (duration, 15 ms; amplitude, approximately 3% of muscle length) caused a faster and larger development of contractile force than in an isometric twitch. The stretch accelerated the decrease of the 1.0 reflection intensity (I1,0). The magnitude of increase of the 1,1 reflection intensity (I1,1) was reduced by the stretch, but its time course was also accelerated. A release applied at the peak of a twitch or later (duration, 5 ms; amplitude, approximately 1.5%) caused only a partial redevelopment of tension. The release produced clear reciprocal changes of reflections toward their relaxed levels, i.e., the I1,0 increased and the I1,1 decreased. A release applied earlier than the twitch peak had smaller effects on the reflection intensities. The results suggest that a strength applied at the onset of a twitch causes a faster radial movement of the myosin heads toward actin, whereas a release applied at or later than the peak of a twitch accelerates their return to the thick filament backbone. The results are discussed in the context of the regulation of the myosin head attachment by calcium.     

Influence of muscle shortening on the geometry of gastrocnemius medialis muscle of the rat

For static and dynamic conditions muscle geometry of the musculus gastrocnemius medialis of the rat was compared at different muscle lengths. The dynamic conditions differed with respect to isokinetic shortening velocity (25, 50 and 75 mm/s) of the muscle-tendon complex and in constancy of force (isotonic) and velocity (isokinetic) during shortening. Muscle geometry was characterized by fibre length and angle as well as aponeurosis length and angle. At high isokinetic shortening velocities (50 and 75 mm/s) small differences in geometry were found with respect to isometric conditions: aponeurosis lengths differed maximally by -2%, fibre length only showed a significant increase (+3.2%) at the highest shortening velocity. The isotonic condition only yielded significant differences of fibre angle (-4.5%) in comparison with isometric conditions. No significant differences of muscle geometry were found when comparing isotonic with isokinetic conditions of similar shortening velocity. The small differences of geometry between isometric and dynamic conditions are presumably due to the lower muscle force in the dynamic condition and the elastic behaviour of the aponeurosis. It is concluded that, unless very high velocities of shortening are used, the relationship between muscle geometry and muscle length in the isometric condition may be used to describe muscle geometry in the dynamic condition.     

ULTRASTRUCTURE OF THE RESTING AND CONTRACTED STRIATED MUSCLE FIBER AT DIFFERENT DEGREES OF STRETCH

Passive stretch, isometric contraction, and shortening were studied in electron micrographs of striated, non-glycerinated frog muscle fibers. The artifacts due to the different steps of preparation were evaluated by comparing sarcomere length and fiber diameter before, during, and after fixation and after sectioning. Tension and length were recorded in the resting and contracted fiber before and during fixation. The I filaments could be traced to enter the A band between the A filaments on both sides of the I band, creating a zone of overlap which decreased linearly with stretch and increased with shortening. This is consistent with a sliding filament model. The decrease in the length of the A and I filaments during isometric contraction and the finding that fibers stretched to a sarcomere length of 3.7 µ still developed 30 per cent of the maximum tetanic tension could not be explained in terms of the sliding filament model. Shortening of the sarcomeres near the myotendinous junctions which still have overlap could account for only one-sixth of this tension, indicating that even those sarcomeres stretched to such a degree that there is a gap between A and I filaments are activated during isometric contraction (increase in stiffness). Shortening, too, was associated with changes in filament length. The diameter of A filaments remained unaltered with stretch and with isometric contraction. Shortening of 50 per cent was associated with a 13 per cent increase in A filament diameter. The area occupied by the fibrils and by the interfibrillar space increased with shortening, indicating a 20 per cent reduction in the volume of the fibrils when shortening amounted to 40 per cent.