The variation of characteristics of twitch and tetanic contractions with sarcomere length in isolated muscle fibres of the frog.

The relation between sarcomere length, tension and time course of tension development in twitch and tetanic contractions at 20 degrees C was determined for isolated fibres from the semitendinosus muscle of the frog (Rana esculenta). In twenty fibres at about 2.15 micron sarcomere length, the peak twitch tension, the maximum tetanic tension and the twitch/tetanus ratio ranged, respectively, from 0.22 to 1.6 kg/cm2, from 2.13 o 3.96 kg/cm2 an from 0.07 to 0.53. The peak twitch tension was found to be: i) directly correlated with the twitch/tetanus ratio and the time to the peak of the first derivative of the twitch tension, ii) inversely correlated with the time to the peak of the first derivative of tetanic tension. No significant correlation was found between the maximal tetanic tension and the peak twitch tension or the twitch/tetanus ratio. Peak twitch tension and twitch/tetanus ratio were not correlated with the fibre cross-sectional area which ranged from 1.052 to 6,283 micron2. Sarcomere length-tension curves for twitch and tetanic isometric contractions at 20 degrees C were determined in twelve fibres. Increases in sarcomere length from about 2.15 to 2.85 micron produced, depending on the peak twitch tension or the twitch/tetanus ratio at about 2.15 micron, either decrease and no change or increase in peak twitch tension, but constantly enhanced the twitch/tetanus ratio and the degree of this potentiation was inversely correlated with the twitch/tetanus ratio at 2.15 micron. Increase in sarcomere length above 2.15 micron did not alter the course of the early development of twitch and tetanic tensions, reduced considerably the variation in peak twitch tension and twitch/tetanus ratio, without altering that of tetanic tension and swamped the correlation between the peak twitch tension and the time to peak of the differentiated twitch tension. However, the peak twitch tension at about 2.85 micron resulted to be directly correlated with the peak twitch tension at about 2.15 micron and in addition the relative length-dependent change in the time of the peak of the first derivative of the twitch tension resulted to be directly correlated with the relative length-dependent change in the peak twitch tension. It is concluded that both the duration of the active state and the rate factors of activation contribute to the determining of the large variation in peak twitch tension at about 2.15 micron, whereas the length-dependent increase in twitch/tetanus ratio appears to be mainly determined by prolongation of the active state duration.     

Estimation of muscle active state

A method is presented for the estimation of the complete time course of muscle active state. The method is based on the selection of a proper model for the muscle, consisting of linear and non-linear components, and on the estimation of its parameters from a simple experiment. The model's parameters are estimated, using the least square method, from measurements of a tetanized muscle's response to a change of its length. The time course of the active state is calculated from an isometric twitch tension response of the same muscle. The twitch tension response is taken as the system's output, and the active state as its input. The latter can be estimated since the system parameters have already been estimated from the tetanized muscle experiment. Experiments were performed on the gastrocnemius muscle of frogs and cats. Results are given for the whole active state time course of these muscles. The results show that the peak active state force does not reach tetanic value, and a negative force is generated during the relaxation period. Additional experiments were carried out with the purpose of verifying the existence of this force; however, no conclusive results were obtained.This research was supported by the Julius Silver Institute of Bio-Medical Engineering Sciences, Grant 050-304     

The motor units of cat medial gastrocnemius: electrical and mechanical properties as a function of muscle length.

The effects of changing muscle length on the mechanical properties of 89 motor units from adult cat medial gastrocnemius have been studied in eight experiments. Few differences were found between the effects of length on tetanic tension, twitch tension, twitch-tetanus ratio, twitch contraction time, twitch half relaxation time, rate of force development and electrical activity for fast contracting (twitch contraction time less than or equal to 45 msec) and slowly contracting (greater than 45 msec) units. Those differences that did appear did not persist when these two groups were matched by tetanic tension. It is concluded that the biophysical mechanisms responsible for the changes in mechanical and electrical properties with length must be similar for fast and slow twitch units and not related to potential differences in their muscle fiber type. The effects of changing muscle length on the mechanical properties of the eight whole muscles suggest that changes in force output with length are of minor importance during normal movements as the muscle is found to be electrically active over a relatively narrow range of lengths close to the optimum length for tetanus of the whole muscle. The very shortest muscle lengths at which there is only minimal force development are not used in natural movements, while the declining limb of the length tension curve is at muscle lengths beyond the maximum in situ length.     

The Pattern of Activation in the Sartorius Muscle of the Frog

The development of isometric twitch tension has been compared with the redevelopment of isometric tension in the fully active frog sartorius muscle following release. At 0°C the rate of rise of isometric twitch tension is the same as that for the muscle in the fully active state at the same tension but not until about 40 msec. after the stimulus and then only for a few milliseconds. The rates of rise of tension in the twitch and in the redevelopment of tension in the fully active muscle appear to be nearly the same at low tensions. Substitution of nitrate for chloride in the Ringer's solution bathing the muscle retards the development of tension during the early part of the contraction phase of the twitch and the effect reaches a maximum within 3 minutes after changing the solutions. These observations have been discussed in connection with some possible patterns of activation and the hypothesis has been advanced that the rate of activation of a sarcomere is determined mainly by the rate at which the transverse component of the link between excitation and contraction is propagated inwards from the periphery to the center of the fiber. This hypothesis has been discussed in relation to others concerning the nature of excitation-contraction coupling.     

Functional morphology of the M. gastrocnemius medialis of the rat during growth

Length-force relations, both active and passive, and twitch contraction characteristics were quantified for left medial gastrocnemius muscles of four young, four adult, and four old male Wistar rats. Muscle and bundle optimum length and muscle weight were also determined and subsequently used for calculation of a number of morphological characteristics of the muscles. Fiber optimum length was derived from muscle bundle optimum length. Generally, physiological characteristics remained constant during growth. There was no change either in active tension at muscle optimum length or in active working range relative to fiber optimum length, relative passive fiber stiffness, active force relative to passive force at optimum length, twitch contraction time and twitch half relaxation time at optimum length. A number of morphological changes, however, did take place in the medial gastrocnemius muscle during growth. Fiber optimum length increased but only by about 2 mm from youth to old age, whereas muscle optimum length increased by approximately 14 mm, presumably owing to extensive hypertrophy of the muscle fibers during growth. The priority for force of the medial gastrocnemius muscle (defined as the quotient of physiological cross-sectional area of a muscle and the cubed root of its volume, a measure independent of architecture and dimensions of muscles) increased during growth. This increase indicates that during growth the muscle shifts relatively more towards force generation than towards excursion generation. These findings are discussed in view of existing scaling theories.     

Active state in heart muscle. Its delayed onset and modification by inotropic agents

The course of active state in heart muscle has been analyzed using a modified quick release method. The onset of maximum active state was found to be delayed, requiring 110-500 msec from time of stimulation, while the time to peak isometric tension required 250-650 msec. Further, the time from stimulation to peak tension was linearly related to the time required to establish maximum intensity of active state as well as to the duration of maximum active state. The duration of maximum active state was prolonged (90-220 msec), occupying most of the latter half of the rising phase of the isometric contraction. Norepinephrine (10(-5) M) shortened the latency from electrical stimulus to mechanical response, accelerated the onset of maximum active state, increased its intensity, decreased its duration, and accelerated its rate of decline. These changes were accompanied by an increase in the rate of tension development and the tension developed while the time from stimulation to peak isometric tension was abbreviated. Similar findings were shown for strophanthidin (1 microgram/ml) although lesser decrements in the duration of maximum active state and time to peak tension were found than with norepinephrine for similar increments in the maximum intensity of active state.     

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.     

Length-dependence of isometric twitch tension potentiation and myosin phosphorylation in mouse skeletal muscle

The effect of changes in muscle length on post-tetanic isometric twitch tension potentiation and myosin P-light chain phosphorylation-was studied at 23°C in the mouse extensor digitorum longus muscle. The length-tension relationship was determined for the same muscles after a 30 min period of quiescence and between 30 s and 3 min after a 1.5 s tetanus at L₀. Isometric twitch tension is increased at all muscle lengths after the tetanus; however, the fractional increase in twitch tension rises from 0.2 at L₀ to a maximum of 0.3 at 1.2 L_0. The fractional increase in twitch tension measured at any fixed muscle length is constant between 30 s and 3 min post-tetanus. P-light chain phosphorylation remains constant between 30 s and 3 min post-tetanus followed by a slow decline to basal values. Under fixed length conditions, there is linear relationship between the relative magnitude of the twitch tension and the extent of P-light chain phosphor-ylation. Net myosin phosphorylalion measured after a 1.5 s tetanus at 1.23 L₀ is 35% less than that obtained under the same conditions at L_0. Thus, contraction-induced phosphorylation of P-light chain decreases with increased muscle length and post-tetanic potentiation at a constant level of P-light chain phosphorylation increases with increasing muscle length. These observations may be consistent with alterations in the sarcoplasmic Ca²⁺ ion transient as the muscle is lengthened.     

Length-dependent twitch contractile characteristics of skeletal muscle

The length dependence of force development of mammalian skeletal muscles was evaluated during twitch, double-pulse, and tetanic contractions, and the relation between muscle length and the time-dependent characteristics of twitch and double-pulse contractions were determined. In situ isometric contractions of the rat gastrocnemius muscle were analyzed at seven different lengths, based on a reference length at which the maximal response to double-pulse contractions occurred (Lopt-2P). Twitch and double-pulse contractions were analyzed for developed tension (DT), contraction time (tC), average rate of force development (DT-tC(-1)), half-relaxation time (t50%R), peak rate of relaxation (DT x dtmin(-1)), and 90%-relaxation time (t90%R). Considering the length at which maximal tetanic DT occurred to be the optimal length (Lo-TET), the peak DT for twitch contractions and double-pulse contractions was observed at Lo-TET + 0.75 mm (p < 0.05) and Lo-TET + 0.1 mm (p > 0.05), respectively. When measured at the length for which maximal twitch and double-pulse contractions were obtained, tetanic DT was 95.2 +/- 3 and 99.0 +/- 2% of the maximal value, respectively. These observations suggest that double-pulse contractions are more suitable for setting length for experimental studies than twitch contractions. Twitch and double-pulse contraction tC were 15.53 +/- 1.14 and 25.0 +/- 0.6 ms, respectively, at Lopt-2P, and increased above Lopt-2P and decreased below Lopt-2P. Twitch t50%R was 12.18 +/- 0.90 ms at Lopt-2P, and increased above Lopt-2P and below Lopt-2P. Corresponding changes for double-pulse contractions were greater. Stretching the muscle leads to slower twitch contractions and double-pulse contractions, but the mechanisms of this change in time course remain unclear.     

The stiffness of slow-twitch muscle lags behind twitch tension: implications for contractile mechanisms and behavior

Small, square stretches were applied during contractions of soleus and plantaris muscles in the cat to measure muscle stiffness. The stiffness of the slow-twitch soleus muscle (but not of the fast plantaris muscle) reaches a maximum after the peak in twitch tension. Since the number of active bonds should be maximum before the peak in tension, we suggest that many bonds are in the rigor state during the falling phase of the twitch. The stiffness of the bonds in this state may be useful for prolonging the twitch in slow-twitch muscles and for maintaining a posture.     

Dynamic properties of cat tenuissimus muscle

Some of the factors which influence the development of tension in cat tenuissimus muscle were studied quantitatively. Under isometric conditions, it was shown that the dynamic properties of the relationship between the tension of the muscle and its electrical stimulation depend on the mean rate of stimulation. This non-linear effect cannot be explained on the basis of the dependence of muscle tension on instantaneous rate of stimulation since the tension due to a stimulus following closely a previous stimulus is augmented, but the time course of the twitch response is unaltered. The interaction between the tension due to active contraction and that due to the viscoelastic properties of the muscle was investigated by independently varying muscle length and the rate of stimulation. Within the limits of resolution of the data, it was concluded that these two components of tension are additive and that muscle stiffness is related to the instantaneous tension of the muscle.     

The peripheral nerve allograft in the primate immunosuppressed with Cyclosporin A: II. Functional evaluation of reinnervated muscle.

Isometric contractile function was evaluated in primates receiving peripheral nerve allografts and autografts. Twelve adult male cynomolgus monkeys received both sural nerve allografts and autografts to the ulnar nerve in opposite forearms. Half the animals received Cyclosporin A (CsA) immunosuppression (25 mg/kg per day); the remaining animals received placebo. One year following nerve engraftment, isometric contractile muscle function was evaluated in reinnervated abductor digiti quinti and intact abductor pollicis brevis muscles. Maximal twitch tension (Pt), tetanic tension (P(o)), time to peak tension (tpt), rate of rise of twitch tension (DP/dt), and muscle fatigue were evaluated at optimal muscle length (L(o)). All reinnervated muscles distal to nerve autografts and allografts in both Cyclosporin A-immunosuppressed and placebo-treated animals generated equivalent maximal twitch tension, tetanic tension, and time to peak tension, with no significant difference between groups (p > 0.05 by ANOVA). There was a tendency toward increased muscle fatiguability in Cyclosporin A-treated animals (p > 0.05). However, the rate of rise of twitch tension was significantly faster in the reinnervated and intact muscles of Cyclosporin A-treated primates (p < 0.05). Evidence of excellent functional reinnervation across nerve allografts and autografts similar to that seen in histologic and electrophysiologic studies was noted. Cyclosporin A immunosuppression did not significantly enhance recovery of muscle function distal to nerve allografts in this model.     

Muscle Volume Changes

Measurements have been made of the volume changes accompanying single isometric and isotonic twitches of frog sartorius muscle. The volume change consists of a rapid increase, a subsequent decrease, and a return to the initial volume; the order of magnitude of increase and decrease is 10^-5 cc/g of muscle. This volume change is length-dependent: the initial increase becomes more pronounced as the initial length of the muscle is decreased, while the volume decrease is greatest at reference length and is diminished for longer and shorter initial lengths. Muscle volume changes are also dependent upon temperature and amount of shortening: the return phase is prolonged as the temperature is lowered; and, in an isotonic twitch, a volume increase accompanying muscle shortening is superimposed upon the volume change described for an isometric twitch. These "shortening volume changes" may correspond to the volume decrease observed in frog muscle under a passive stretch. If the active state is prolonged by the use of a frog Ringer solution in which iodide ions have been substituted for chloride ions, the time course of the volume decrease is likewise prolonged; this suggests a relationship between the volume decrease and the active state of the muscle.     

Mechanical properties of denervated amphibian muscle

Denervated amphibian muscle does not show the prolongation of action potential found in mammalian denervated muscle. It was, therefore, predicted that denervated amphibian muscle would not show prolongation of the mechanical twitch. The sartorius muscles in one leg of toads--Xenopus borealis--were denervated for 140-268 days. Isometric twitch time to peak, time to half relaxation and twitch/tetanus ratio were not changed following denervation, confirming our prediction. Twitch tension decreased to 68% and tetanic tension decreased to 75% of control values. The maximum velocity of unloaded shortening (muscle length/s) was also unchanged.     

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.     

A data-acquisition system for the analysis of the isometric tension generated by an electrically stimulated skeletal muscle

The system is designed for data-acquisition and computationof a relatively large nwnber of parameters of a single twitchor tetanic contraction generated by an electrically stimulatedskeletal muscle. The parameters that can be computed using theprogram facilities are: (i) the rise time (T_c), 50% relaxationtime (50% T_r) and the maximwn tendon (P_t) of the twitch contraction;(ii) the approximate number of motor units innervating the muscleand the fatigue index; (iii) the fission frequency of the tetaniccontraction and the twitch to tetanus ratio; (iv) the maximumtetanic tension (P₀) and its T_c and 50% T_n (v) the P₀ versusmuscle length diagram; (vi) the fatigue-time course.     

The active tension-length curve of vascular smooth muscle related to its cellular components

The active and passive isometric tension-length (internal circumference) relation of vascular smooth muscle has been investigated using a 100-200-micron lumen diameter artery from the rat mesenteric bed. Conditions were established under which maximal activation was obtained at all lengths. Below L0 (the length at which maximum tension, delta T0, was developed) the active tension fell with decreasing length along a line which extrapolated to 0.38 L0; below 1.1 L0 the relation was reversible regardless of the protocol used. Above L0 the active tension fell linearly with increasing length along a line which extrapolated to zero tension at 1.82 L0. At the longer lengths investigated (up to 1.6 L0) the passive tension upon which the active responses were superimposed was as high as 4.4 delta T0. However, measurements of the dynamic characteristics of the preparation (with a time resolution of 2 ms) suggest that the active tension measured is nevertheless a measure of the active properties of the contractile apparatus. Direct light microscopic observation of the effect of length change on the cells within the walls of the preparation showed that changes in vessel length produced, on average, the same percentage change in cell length. Histological examination showed no signs of cell destruction following large extensions. The results suggest that the decrease in tension with extension above L0 is due to changes in the properties of the contractile apparatus, rather than to cellular damage.     

Time-resolved X-ray diffraction studies of frog skeletal muscle isometrically twitched by two successive stimuli using synchrotron radiation

In order to clarify the delay between muscular structural changes and mechanical responses, the intensity changes of the equatorial and myosin layer-line reflections were studied by a time-resolved X-ray diffraction technique using synchrotron radiation. The muscle was stimulated at 12-13 degrees C by two successive stimuli at an interval (80-100 ms) during which the second twitch started while tension was still being exerted by the muscle. At the first twitch, the intensity changes of the 1.0 and 1.1 equatorial reflections reached 65 and 200% of the resting values, and further changes to 55 and 220% were seen at the second twitch, respectively. Although the second twitch decreased not only the time to peak tension but also that to the maximum intensity changes of the equatorial reflections (in both cases, about 15 ms), the delay (about 20 ms) between the intensity changes and the development of tension at the first twitch were still observed at the second twitch. On the other hand, the intensities of the 42.9 nm off-meridional and the 21.5 nm meridional myosin reflections decreased at the first twitch to the levels found when a muscle was isometrically tetanized, and no further decrease in their intensities was observed at the second twitch. These results indicate that a certain period of time is necessary for myosin heads to contribute to tension development after their arrival in the vicinity of the thin filaments during contraction.     

Sensitivity of cultured and skinned chick myotube to calcium, strontium, and barium ions examined by recording isometric contractions.

Sensitivity of cultured chick myotubes to alkaline earth metal ions was investigated by recording contractile isometric tension through a semiconductor transducer. The myotubes were obtained by culturing myoblasts of chick embryo breast muscles, and skinned chemically before physiological experiments. Contractions developed in response to Ca2+ in a bathing medium higher than 3 x 10(-7) M and reached maximum at 1 x 10(-5) M. Sr2+ was less effective than Ca2+; the threshold concentration was 1 x 10(-5) M and the tension reached maximum at 1 x 10(-3) M. Ba2+ was the least effective among the three alkaline earth metal ions; only one fifth of the Ca(2+)-induced maximum tension was attained at 1 x 10(-3) M. The sensitivity was similar to that of the mature pectoral muscle fiber, a fast twitch muscle fiber, rather than that of the anterior latissimus dorsi, a slow tonic muscle fiber. The sensitivity was shown to be dependent on its troponin C by replacing it with troponin C from the mature pectoral or cardiac muscle. This indicates that TnC of a fast-muscle type is expressed in the cultured chick myotube as in the mature pectoral muscle. The contractile apparatus was thus shown to be well developed in the cultured myotube with characteristics similar to the mature fast twitch muscle fiber.     

Thermal adaptations in lizard muscle function

Summary This study was undertaken to investigate thermal adaptations in muscle contractile properties in closely-related lizards with different preferred body temperatures (PBT). The species examined all belong to theSphenomorphus group of Australian skinks (Scincidae: Lygosominae). Preferred body temperatures are conservative at the generic level as follows:Ctenotus, 35°C;Sphenomorphus, 30°C;Eremiascincus, 25°C. Contractile properties of the fast glycolytic portion of the iliofibularis muscle were measured. Translational adaptations are evident in several isometric factors, including tetanic tension (Po), twitch tension (Pt), twitch time to peak tension (TPT), and twitch half-relaxation time (1/2 RT). Capacity adaptations are not evident in rates of tetanic tension development (dPo/dt) or in maximal velocities of isotonic shortening (V _max). Rotational adaptations are not evident in any contractile properties. Thermal limits on upper response temperatures are about 5°C warmer inCtenotus than in the more cryophilic species, indicative of resistance adaptation in muscle performance. Despite these adaptive shifts, there is little indication that muscle functional capacities are optimized or equalized at PBT in these lizards.Abbreviations FG fast glycolytic - IF iliofibularis muscle - PBT preferred body temperature - Po tetanic tension - Pt twitch tension - 1/2RT twitch half relaxation time - TPT twitch time to peak tension