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Energetics of cardiac contractions.

1. The average resting heat production of a muscle under zero tension is 24.8 mcal/g muscle .min at 20 degrees C. In the majority of muscles examined the resting heat production increases when the resting tension and muscle length are increased.2. The relation between actively developed tension and heat produced is similar to that existing in skeletal muscle. The plot of heat against developed tension can be obtained either by altering muscle length or by varying the stimulus frequency.3. The mean maximum total efficiency work/(work + heat) in the work experiments was 11.6%. The total energy produced (work + heat) correlates with the load rather than with the work done.4. In isotonic contractions more heat is liberated than the heat versus tension plot predicts. This extra heat is load-dependent.

Animals↗

Contraction kinetics of intact and skinned frog muscle fibers and degree of activation. Effects of intracellular Ca2+ on unloaded shortening.

This study addresses a long-standing controversy on the effects of the degree of activation on cross-bridge kinetics in vivo, by utilizing isolated intact and skinned fiber preparations. Steady force levels ranging from 0.1 to 0.76 P0 were achieved at 0 degrees C with temperature-step stimulation of intact fibers by varying the amount of caffeine in the bathing medium. The speed of unloaded shortening (by slack test) was found to be practically constant, which suggests that intracellular Ca2+ in the intact preparation has relatively little effect on isotonic shortening. Along with the results on tetanically stimulated fibers (force, P0), we observed a minor but significant trend for the speed to decline with lowered force levels. This trend is explained by the presence of a constant internal load equaling approximately 1% P0. The effect of Ca2+ on the shortening behavior of skinned fibers was examined at 0 and 10 degrees C. At 0 degrees C, there was practically no effect of Ca2+ on the shortening response in slack tests. At 10 degrees C, there was also no Ca2+ effect during the first activation cycle, but in subsequent cycles the speed of shortening was reduced during partial activation, which indicates that there were permanent changes in the fiber properties under these experimental conditions. The latter result could be explained if the internal load had increased to approximately 5% P0 in the modified skinned fiber (compared with 1% P0 in intact fiber). These findings show that isotonic contraction of frog fibers is intrinsically unaffected by the variations in intracellular Ca2+ that modulated the force over a nearly complete range. The results provide support for the idea that Ca2+ influences the force development in vivo by on-off switching mechanisms.

Animals↗

Force velocity relationships in vascular smooth muscle. The influence of temperature.

Force velocity relationships of isolated vascular smooth muscle preparations were examined in the tetanized rat portal--anterior mesenteric vein by means of afterloaded isotonic contractions. Lowering of the temperature from 37 degrees C to 25 degrees C caused an average decrease of the follwoing parameters: maximal velocity of shortening at zero load from 0.51 to 0.28 muscle length/sec; maximal rate of tension increase (dT/dt) from 847 to 362 dyn/sec; mechanical tension developed at the maximum of dT/dt from 43 to 34% of the peak force generation; constant b of Hill's equation from 0.18 to 0.09 muscle length/sec. The latency was prolonged from 0.33 to 0.56 sec, and the time-to-maximum of dT/dt from 0.9 to 1.6 sec. Between 27 degrees C and 37 degrees C the following Q10 values were calculated: for the maximal velocity of shortening at zero load 1.56; for the maximal rate of tension increase 1.88; for the latency 1.63. No distinct influence of temperature could be observed on the extent of isotonic shortening at zero load (3.69 muscle length at 37 degrees C), on the extent of peak force generation (1107 dyn at 37 degrees C), and on constant a of Hill's equation (0.35 at 37 degrees C). It is concluded that parameters of contraction velocity in vascular smooth muscle depend on temperature, whereas the extent of contraction is independent of temperature. These findings are discussed in connection with the theory of the sliding-filament mechanism.

Animals↗

Maturational changes in troponin T expression, Ca2+-sensitivity and twitch contraction kinetics in dragonfly flight muscle

Maximum lift production and the thermal sensitivity of lift production increase dramatically during adult maturation of Libellula pulchella dragonflies. Here, we report that the mechanistic basis for this transition appears to involve a developmental change in protein expression, which alters the Ca2+-sensitivity of muscle activation and twitch contraction kinetics. The alternatively spliced Ca2+ regulatory protein troponin T (TnT) undergoes an isoform shift during adult maturation. Skinned (demembranated) fibers of mature flight muscle are up to 13 times more sensitive to activation by Ca2+ than skinned fibers from teneral (newly emerged adult) flight muscle, and their Ca2+-sensitivity is more strongly affected by temperature. Intact muscle from mature individuals has a shorter time to peak tension and longer time to half-relaxation during twitch contractions, which is consistent with a greater Ca2+-sensitivity of mature muscle. Because it becomes activated more quickly and relaxes more slowly, mature flight muscle is able to generate, with each twitch, more force per unit area than teneral muscle; this difference in force becomes greater at high temperatures. There do not appear to be any age-related differences in actomyosin crossbridge properties, since teneral and mature flight muscles do not differ in shortening velocity, tetanic tension or instantaneous power output during isotonic contraction. Thus, variation in TnT expression appears to affect the temperature-dependent Ca2+-sensitivity of muscle activation, which in turn affects the kinetics and force production of the twitch contractions used by dragonflies during flight. This cascade of effects suggests that maturational changes in the expression of TnT isoforms may be a key determinant of overall muscle and organismal performance.

Journal Article↗

Changes in intracellular calcium during mechanical alternans in isolated ferret ventricular muscle.

Alternans in heart is important as pulsus alternans in cardiac failure and electrophysiological alternans in myocardial ischemia. The explanation of this phenomenon is still unclear. We attempted to investigate the cellular mechanisms of alternans by measuring intracellular free calcium concentration [( Ca2+]i) with the photoprotein aequorin in isolated ferret papillary muscles. Tension and length were also recorded simultaneously. Transient mechanical alternans lasting five to 20 contractions could be reliably induced in this preparation by following a 30-second rest period with stimulation at a fast rate (2-4 Hz). Production of sustained mechanical alternans, which lasted longer than 20 contractions and could persist for several hundred contractions, required additional interventions, consisting of a lower temperature (25 degrees C), a lower external calcium concentration (1 mM), and a lower pH (6.91) than control conditions (0.33-0.5 Hz, 30 degrees C, 2 mM Ca2+, pH 7.36). Transient mechanical alternans was associated with transient in-phase alternation of aequorin light and, hence, [Ca2+]i. Sustained mechanical alternans was associated with sustained in-phase alternation of aequorin light as well as incomplete relaxation of tension. However, when muscles were switched from isometric to unloaded isotonic contraction, relaxation between stimuli was complete but contraction and the aequorin light signal continued to alternate. The addition of 10 mM caffeine or 10 microns ryanodine abolished transient and sustained mechanical alternans and also abolished the associated alternation of aequorin light. Commensurate with the action of ryanodine, which allows the sarcoplasmic reticulum to reaccumulate calcium to a limited extent after a period of rapid stimulation, sustained mechanical alternans sometimes reappeared in an attenuated form 30 to 50 contractions after the addition of ryanodine. These results demonstrate that incomplete muscle relaxation between beats need not be present for alternans to occur, and support the hypothesis that alternans is caused by intracellular calcium cycling involving the sarcoplasmic reticulum.

Aequorin↗

The Anrep effect: an intrinsic myocardial mechanism.

In cat papillary muscles contracting physiologically, increasing the afterload caused a biphasic change in contractility. In response to an increase in afterload, contractility (as measured by peak shortening, peak developed force, or peak dF/dt) initially decreased (antihomeometric autoregulation) over the first few beats and then increased slowly with t 1/2 of about 3 min at 30 degrees C and about 1 min at 37 degrees C (homeometric autoregulation). The antihomeometric autoregulation is due to decreased active shortening when the afterload is increased, since it also occurs in response to increased afterload in isotonic contractions. The secondary slow increase in contractility is primarily due to the increase in mean diastolic length that occurs as a result of increased afterload. The time course and the magnitude of the biphasic change in contractility are very similar to those observed in response to afterload increase in intact hearts; we suggest that the secondary slow increase in contractility that we observed is a contributory mechanism to homeometric autoregulation (or the Anrep effect), as it is observed in the whole heart.

Animals↗

Muscarine receptor types mediating autoinhibition of acetylcholine release and sphincter contraction in the guinea-pig iris.

The potencies of several muscarine receptor antagonists in blocking either the autoinhibition of acetylcholine release or the muscarinic contraction of the sphincter muscle upon acetylcholine release were investigated in the guinea-pig iris. The agonist at pre- or postjunctional muscarine receptors was acetylcholine released upon field stimulation (5.5 Hz, 2 min) of the irides preloaded with 14C-choline. The stimulation-evoked 14C-overflow was doubled in the presence of atropine 0.1 mumol/l but unaffected by the agonist (+/-)-methacholine (50 mumol/l). Thus, under the present stimulation conditions, the autoinhibition of acetylcholine release on the guinea-pig iris cholinergic nerves was nearly maximally activated. Isotonic contractions of the irides upon field stimulation consisted of a rapid, atropine (0.1 mumol/l)-sensitive peak phase followed by a sustained contraction which involved a cholinergic and a non-cholinergic stimulation of the sphincter muscle. The M2-selective antagonists methoctramine (10 mumol/l) and gallamine (100 mumol/l) increased both the 14C-overflow and the peak contractions evoked by field stimulation. In contrast, the M3-selective antagonist hexahydrosiladifenidol (0.1-10 mumol/l) failed to affect the evoked 14C-release but concentration-dependently (1-10 mumol/l) reduced the iris contractions. Pirenzepine (10 mumol/l) enhanced the evoked 14C-overflow and inhibited the peak contractions (0.1-10 mumol/l; maximal effect at 10 mumol/l). The low potency of the antagonist at both receptor sites indicates that an M1 muscarine receptor is not involved. The results are consistent with the idea of M2 muscarine receptors mediating autoinhibition of acetylcholine release in the guinea-pig iris and M3-like receptors inducing the contraction of the sphincter muscle.

Acetylcholine↗

Compression of EMG signals with wavelet transform and artificial neural networks.

This paper presents a hybrid adaptive algorithm for the compression of surface electromyographic (S-EMG) signals recorded during isometric and/or isotonic contractions. This technique is useful for minimizing data storage and transmission requirements for applications where multiple channels with high bandwidth data are digitized, such as telemedicine applications. The compression algorithm proposed in this work uses a discrete wavelet transform for spectral decomposition and an intelligent dynamic bit allocation scheme implemented by an approach using the Kohonen layer, which improves the bit allocation for sections of the S-EMG with different characteristics. Finally, data and overhead information are packed by entropy coding. The results for the compression of isometric EMG signals showed that this algorithm has a better performance than standard wavelet compression algorithms presented in the literature (presenting a decrease of at least 5% in per cent residual difference (PRD) for the same compression ratio), and a performance that is comparable with the performance of algorithms based on an embedded zero-tree wavelet. For isotonic EMG signals, its performance is better than the performance of the algorithms based on embedded zero-tree wavelets (presenting a decrease in PRD of about 3.6% for the same compression ratios, in the useful compression range).

Algorithms↗

Magnesium ion-dependent contraction of skinned frog muscle fibers in calcium-free solution.

Skinned frog fibers were reversibly activated in Ca-free solutions containing 0 mM KCl, 23 microM free Mg, and having an ionic strength of approximately 50 mM. Contractile force was nearly maximal at 22 degrees - 25 degrees C and decreased at lower temperatures. Maximal force in Ca-free solution at 50 mM ionic strength was close to twice the calcium-activated force with pCa 5 and 190 mM ionic strength. The force in Ca-free solution could be reduced to zero by raising the concentration of free Mg from 23 microM to 1.0 mM at the same ionic strength (50 mM). On stretching the fiber from 2.0 to 3.2 micron the force decreased; this effect was similar to that seen with Ca-activated fiber and the data support the idea that Ca-free tension is made at the cross-bridge level. Isotonic contraction during Ca-free activation showed a velocity transient as in Ca-activated fiber at 190 mM ionic strength, but the transient in the present case was very much prolonged. This finding suggests that contraction mechanisms for force generation and for shortening are essentially the same in the two conditions, but that certain rate constants of cross-bridge turnover are slower for the Ca-free contraction. Also, the results indicate that, in low ionic strength, Ca binding to thin filaments is not essential for unmasking the cross-bridge attachment sites, which suggests that the steric blocking mechanism is modified under these conditions.

Animals↗

Kinetics of thin filament activation probed by fluorescence of N-((2-(iodoacetoxy)ethyl)-N-methyl)amino-7-nitrobenz-2-oxa-1,3-diazole-labeled troponin I incorporated into skinned fibers of rabbit psoas muscle: implications for regulation of muscle contraction.

Making use of troponin with fluorescently labeled troponin I subunit (N-((2-(iodoacetoxy)ethyl)-N-methyl)amino-7-nitrobenz-2-oxa-1, 3-diazole-troponin I, IANBD-TnI) that had previously been described in solution studies as a probe for thin filament activation (. Proc. Natl. Acad. Sci. 77:7209-7213), we present a new approach that allows the kinetics of thin filament activation to be studied in skinned muscle fibers. After the exchange of native troponin for fluorescently labeled troponin, the fluorescence intensity is sensitive to both changes in calcium concentration and actin attachment of cross-bridges in their strong binding states (. Biophys. J. 77:000-000). Imposing rapid changes in the fraction of strongly attached cross-bridges, e.g., by switching from isometric contraction to high-speed shortening, causes changes in thin filament activation at fixed Ca(2+) concentrations that can be followed by recording fluorescence intensity. Upon changing to high-speed shortening we observed small (<20%) changes in fluorescence that became faster at higher Ca(2+) concentrations. At all Ca(2+) concentrations, these changes are more than 10-fold faster than force redevelopment subsequent to the period of unloaded shortening. We interpret this as an indication that equilibration among different states of the thin filament is rapid and becomes faster as Ca(2+) is raised. Fast equilibration suggests that the rate constant of force redevelopment is not limited by changes in the activation level of thin filaments induced by the isotonic contraction before force redevelopment. Instead, our modeling shows that, in agreement with our previous proposal for the regulation of muscle contraction, a rapid and Ca(2+)-dependent equilibration among different states of the thin filament can fully account for the Ca(2+) dependence of force redevelopment and the fluorescence changes described in this study.

Actin Cytoskeleton↗

Is there a shortening-heat component in mammalian cardiac muscle contraction?

It has been suggested that there is a shortening-heat component that is an extra liberation of heat on shortening above that due to the external work, which contributes to the total energy expenditure of the beating heart. The presence of a shortening heat component was studied in isolated papillary muscles from the right ventricle of rabbits killed by cervical dislocation. At the onset of a contraction, muscles were shortened from various initial lengths through fixed distances at near maximum velocity before being allowed to develop force at the new length; the heat production accompanying such contractions was measured. The measured heat was compared with heat values predicted from previously established heat-stress curves obtained by using either preshortening or latency release methods. There was no shortening-related increment in heat output per contraction when comparison was made to a control heat-stress curve, obtained using the latency release method. An increase in heat production of 10% was observed with long shortening distances when comparison was made to a control heat-stress curve obtained by preshortening the muscles; however, this difference is most likely due to an underestimate of the magnitude of the activation heat component in these control heat-stress curves. An increase in isometric heat production due to maintained stretch per se was observed. The present data indicate that it is unlikely that there is a significant shortening heat component when cardiac muscle shortens. The absence of such a metabolic component may account for the rapid fall off in total enthalpy output in isotonic contractions at low to medium afterloads when compared with the skeletal muscle data.

Analysis of Variance↗

Effect of duration of vagal stimulation on shortening velocity of in vivo canine trachealis muscle.

We studied shortening velocity of in vivo canine trachealis muscle contracted by bilateral vagus nerve stimulation, as a function of duration of contraction. The cervical trachea was transected at two locations, cut at the ventral portion, and opened. One side of the cartilage cut was connected to a force transducer and the other to a lever with a given weight as an afterload, the length of which was measured by a linear displacement transducer. Bilateral vagosympathetic trunks were stimulated by supramaximal electrical impulses. With vagal stimulation, the trachealis muscle started to contract isometrically and at a given time the muscle was allowed to contract isotonically by unlocking a stopper at a given afterload. The shortening velocity was reduced with longer duration of active state. Ten sec after vagal stimulation the maximum force was 730 +/- 105 g/cm2 (mean +/- S.D.) and maximum velocity at zero load calculated by Hill's equation was 0.092 l0/sec. We conclude that the force-velocity relationship of in vivo canine trachealis muscle stimulated by vagus nerves exhibits a time-dependency similar to that in vitro.

Animals↗

Comparison of Caplan's irreversible thermodynamic theory of muscle contraction with chemical data.

Recently Caplan (1) applied the concepts of irreversible thermodynamics and cybernetics to contracting muscle and derived Hill's force-velocity relation. Wilkie and Woledge (2) then compared Caplan's theory to chemical rates inferred from heat data and concluded that the theory was not consistent with the data. Caplan defended his theory in later papers (3, 4) but without any direct experimental verifications. As Wilkie and Woledge (2) point out, the rate of phosphorylcreatine (PC) breakdown during steady states of shortening has not been observed because of technical difficulties. In this paper it is shown that the rate equations may be directly integrated with time to obtain relations among actual quantities instead of rates. The validity of this integration is based on experimental evidence which indicates that certain combinations of the transport coefficients are constant with muscle length. These equations are then directly compared to experimental data of Cain, Infante, and Davies (5) with the following conclusions: (a) The measured variations of DeltaPC for isotonic contractions are almost exactly as predicted by Caplan's theory. (b) The value of the chemical rate ratio, nu(m)/nu(o), obtained from these data was 3.53 which is close to the value of 3 suggested by Caplan (3). (c) The maximum value of the chemical affinity for PC splitting was found to be 10.6 k cal/mole which is as expected from in vitro measurements (2). Because of the excellent agreement between theory and experiment, we conclude that Caplan's theory definitely warrants further investigation.

Animals↗

Theory of muscular contraction extended to groups of actin sites.

It was shown in an earlier paper how to connect, in principle, the biochemical states of a cross-bridge with the mechanics of muscular contraction, by the methods of statistical mechanics. The treatment applies to cross-bridges that are able to interact with only one actin site at a time. The present paper shows that it is a straightforward matter to extend the theory to groups of actin sites (three, five, etc.), say 55 A apart, as suggested by the work of Moore, H. E. Huxley, and DeRosier. The possibility of the cross-bridge attachment slipping between sites is included. This provides an alternative molecular interpretation of the model introduced by A. F. Huxley and Simmons. A second possible interpretation is also suggested: their discrete stable angles correspond to different biochemical (attached) states. The Huxley-Simmons analysis of an example is rederived and extended somewhat (x averaging), from the point of view of the present theory. Their qualitative conclusions are left unchanged by the x averaging, but significant quantitative effects are possible. Possible consequences of fast slipping in isotonic contraction are discussed in a preliminary way.

Actins↗

The rate of resting heat production of rat papillary muscle.

The rate of resting heat production of quiescent rat left ventricular papillary muscles was measured myothermically. The effects of contractile activity, stretch, oxygen partial pressure, temperature, amino acids and time were examined. The rate of basal heat production was the same throughout the day whether or not muscles contracted isotonically under a small pre-load. Passive stretch increased the rate of resting heat production; the stretch-induced increment was highly variable from muscle to muscle. The resting heat rate per se was only moderately sensitive to oxygen partial pressure and temperature, and was insensitive to the presence of amino acids in the bathing medium. The stretch-induced increase in resting heat rate was independent of these three factors. The rate of resting heat production declined exponentially with time to reach a plateau about 4 h following cardiectomy.

Amino Acids↗

Viscoelastic properties of the contracting detrusor. II. Experimental approach.

Mechanical properties of detrusor muscle were studied with small-amplitude oscillatory volume perturbations in isometrically contracting bladders of anesthetized dogs. Contractions were studied at oscillatory frequencies (f) of 2 and 4 Hz and at bladder volumes (Vbl) ranging from 30 to 110 ml. The magnitude of bladder hydrodynamic stiffness (magnitude of G) increased linearly with mean detrusor pressure (Pdet) while the phase angle remained relatively constant during contraction. The slope (mG) of magnitude of G-Pdet relations had a positive dependence on f and a negative dependence on Vbl. Analysis of oscillatory data, described in the companion paper, was performed using incremental lumped-parameter models consisting of a spring with incremental constant (S = dF/dL), a viscous element with incremental viscosity (b = dF/du), and a mass (m). Only the model where elastic and viscous elements were placed in series with each other and in parallel with mass was compatible with the experimental data. Both S and b increased linearly with effective force (F), defined as Pdet times the cross-sectional area of the intravesical cavity. Slopes of the S-F and b-F relationships (ms and mb) were independent of Vbl and varied only slightly with f. The importance of this finding stems from recognizing that ms and mb correspond to the exponential coefficients of nonlinear series elastic and internal viscosity elements. These parameters, when normalized by resting muscle length, represent fundamental muscle properties independent of muscle cross-sectional area, stretch, or level of activation and compare well with parameters derived from other muscle systems using techniques such as quick releases and isotonic contractions.

Animals↗

Spectral analysis of the electromyograph of the erector spinae muscle before and after a dynamic manual load-lifting test.

The aim of the present study was to assess the spectral behavior of the erector spinae muscle during isometric contractions performed before and after a dynamic manual load-lifting test carried out by the trunk in order to determine the capacity of muscle to perform this task. Nine healthy female students participated in the experiment. Their average age, height, and body mass (+/- SD) were 20 +/- 1 years, 1.6 +/- 0.03 m, and 53 +/- 4 kg, respectively. The development of muscle fatigue was assessed by spectral analysis (median frequency) and root mean square with time. The test consisted of repeated bending movements from the trunk, starting from a 45 masculine angle of flexion, with the application of approximately 15, 25 and 50% of maximum individual load, to the stand up position. The protocol used proved to be more reliable with loads exceeding 50% of the maximum for the identification of muscle fatigue by electromyography as a function of time. Most of the volunteers showed an increase in root mean square versus time on both the right (N = 7) and the left (N = 6) side, indicating a tendency to become fatigued. With respect to the changes in median frequency of the electromyographic signal, the loads used in this study had no significant effect on either the right or the left side of the erector spinae muscle at this frequency, suggesting that a higher amount and percentage of loads would produce more substantial results in the study of isotonic contractions.

Adult↗

Inhibition of the p38 MAP kinase pathway destabilizes smooth muscle length during physiological loading.

We tested the hypothesis that mechanical plasticity of airway smooth muscle may be mediated in part by the p38 mitogen-activated protein (MAP) kinase pathway. Bovine tracheal smooth muscle (TSM) strips were mounted in a muscle bath and set to their optimal length, where the active force was maximal (F(o)). Each strip was then contracted isotonically (at 0.32 F(o)) with ACh (maintained at 10(-4) M) and allowed to shorten for 180 min, by which time shortening was completed and the static equilibrium length was established. To simulate the action of breathing, we then superimposed on this steady distending force a sinusoidal force fluctuation with zero mean, at a frequency of 0.2 Hz, and measured incremental changes in muscle length. We found that TSM strips incubated in 10 microM SB-203580-HCl, an inhibitor of the p38 MAP kinase pathway, demonstrated a greater degree of fluctuation-driven lengthening than did control strips, and upon removal of the force fluctuations they remained at a greater length. We also found that the force fluctuations themselves activated the p38 MAP kinase pathway. These findings are consistent with the hypothesis that inhibition of the p38 MAP kinase pathway destabilizes muscle length during physiological loading.

Animals↗