Calcium efflux during the cold-induced contraction of mammalian striated muscle fibres

The efflux of ⁴⁵Ca from mammalian slow twitch muscle fibres has been studied to provide a measure of the concentration of free Ca²⁺ in the sarcoplasm. The kinetically complex early phases of washout of the isotope are succeeded by a prolonged slower phase which exhibits first-order kinetics. This later phase is accelerated by caffeine, by preventing oxidative phosphorylation and also during an isometric contraction, whether this contraction is produced by lowering the temperature or by electrical stimulation. The local anaesthetic tetracaine abolishes the contraction caused by cold and in this case the rate constant for efflux is progressively lowered as the temperature is reduced (Q₁₀ value of 2.3). The removal of external Na⁺ and Ca²⁺ reduces the efflux rate constant. Caffeine, sodium removal and the inhibition of oxidative phosphorylation, all potentiate the cold contraction and the associated extra ⁴⁵Ca efflux. Ca removal causes the cold contraction to become phasic. It appears that caffeine, sodium removal, the inhibition of oxidative phosphorylation and a decrease in temperature to below 10°C are all treatments which, like electrical stimulation, increase the sarcoplasmic free calcium concentration to varying degrees.     

Effect of deuterium oxide on contraction characteristics and ATPase activity in glycerinated single rabbit skeletal muscle fibers

We studied the effect of deuterium oxide (D₂O) on contraction characteristics and ATPase activity of single glycerinated muscle fibers of rabbit psoas. D₂O increased the maximum isometric force P₀ by about 20%, while the force versus stiffness relation did not change appreciably. The maximum shortening velocity under zero load V_max did not change appreciably in D₂O, so that the force-velocity (P-V) curve was scaled depending on the value of P₀. The Mg-ATPase activity of the fibers during generation of steady isometric force P₀ was reduced by about 50% in D₂O. Based on the Huxley contraction model, these results can be accounted for in terms of D₂O-induced changes in the rate constants f₁ and g₁ for making and breaking actin-myosin linkages in the isometric condition, in such a way that f₁/(f₁+g₁) increases by about 20%, while (f₁+g₁) remains unchanged. The D₂O effect at the molecular level is discussed in connection with biochemical studies on actomyosin ATPase.     

Inferring crossbridge properties from skeletal muscle energetics

Work is generated in muscle by myosin crossbridges during their interaction with the actin filament. The energy from which the work is produced is the free energy change of ATP hydrolysis and efficiency quantifies the fraction of the energy supplied that is converted into work. The purpose of this review is to compare the efficiency of frog skeletal muscle determined from measurements of work output and either heat production or chemical breakdown with the work produced per crossbridge cycle predicted on the basis of the mechanical responses of contracting muscle to rapid length perturbations. We review the literature to establish the likely maximum crossbridge efficiency for frog skeletal muscle (0.4) and, using this value, calculate the maximum work a crossbridge can perform in a single attachment to actin (33 × 10⁻²¹ J). To see whether this amount of work is consistent with our understanding of crossbridge mechanics, we examine measurements of the force responses of frog muscle to fast length perturbations and, taking account of filament compliance, determine the crossbridge force-extension relationship and the velocity dependences of the fraction of crossbridges attached and average crossbridge strain. These data are used in combination with a Huxley-Simmons-type model of the thermodynamics of the attached crossbridge to determine whether this type of model can adequately account for the observed muscle efficiency. Although it is apparent that there are still deficiencies in our understanding of how to accurately model some aspects of ensemble crossbridge behaviour, this comparison shows that crossbridge energetics are consistent with known crossbridge properties.     

Thin filament flexibility and its role in muscle contraction

Contemporary experimental methods do not allow unequivocal determination of molecular structural events during muscle contraction. To analyze existing contradictions, an original computer program has been developed. This program reconstructs the hexagonal lattice of a sarcomere for different states of muscle and finds the most realistic structure by comparing the calculated Fourier spectrum with the actual diffraction pattern. Previously, the new approach allowed reconstructing the actual structure of a myosin filament from mammalian striated muscle (http://zope.ibib.waw.pl/pspk). In this work, the thin filament is reconstructed for three states: relaxed, activated, and contracting. The good fit between the calculated Fourier spectra and the actual diffraction patterns taken from the literature suggests that the thin filament owing to its flexibility may play an active role in muscle contraction, as myosin cross-bridges do.     

Changes in the lateral filament spacing of skinned muscle fibres when cross-bridges attach

When a skinned fibre prepared from frog skeletal muscle goes from the relaxed to the rigor state at a sarcomere length of about 2.2 μm, the 1, 0 transverse spacing of the filament lattice, measured by X-ray diffraction, decreases by about 11%. In measurements at various sarcomere lengths, the decrease in the spacing was approximately proportional to the degree of overlap between the thick and thin filaments. This suggests that the shrinkage of the lattice is caused by a lateral force produced by cross-bridges. In order to estimate the magnitude of the lateral force, the decrease of spacing between relaxed and rigor states was compared with the shrinkage caused osmotically by adding a high molecular weight polymer, polyvinylpyrrolidone, to the bathing solution. The results indicate that the lateral force produced per unit length of thick filament in the overlap zone is of the same order of magnitude as the axially directed force produced during maximum isometric contraction (10^?10 to 10^?9 N/μm).Experiments in the presence of a high concentration of polyvinylpyrrolidone (100 g/l) show that when the lattice spacing is decreased osmotically beyond a certain value, the lateral force produced when the fibre goes into rigor changes its direction, causing the lattice to swell. This result can be explained by assuming that there is an optimum interfilament spacing at which the cross-bridges produce no lateral force. At other spacings, the lateral force tends to displace the filament lattice toward that optimum value.     

Considerations on muscle contraction

The independent force generator and the power-stroke cross-bridge model have dominated the thinking on mechanisms of muscular contraction for nearly the past five decades. Here, we review the evolution of the cross-bridge theory from its origins as a two-state model to the current thinking of a multi-state mechanical model that is tightly coupled with the hydrolysis of ATP. Finally, we emphasize the role of skeletal muscle myosin II as a molecular motor whose actions are greatly influenced by Brownian motion. We briefly consider the conceptual idea of myosin II working as a ratchet rather than a power stroke model, an idea that is explored in detail in the companion paper.     

Calcium indicators and calcium signalling in skeletal muscle fibres during excitation–contraction coupling

During excitation–contraction coupling in skeletal muscle, calcium ions are released into the myoplasm by the sarcoplasmic reticulum (SR) in response to depolarization of the fibre’s exterior membranes. Ca²⁺ then diffuses to the thin filaments, where Ca²⁺ binds to the Ca²⁺ regulatory sites on troponin to activate muscle contraction. Quantitative studies of these events in intact muscle preparations have relied heavily on Ca²⁺-indicator dyes to measure the change in the spatially-averaged myoplasmic free Ca²⁺ concentration (Δ[Ca²⁺]) that results from the release of SR Ca²⁺. In normal fibres stimulated by an action potential, Δ[Ca²⁺] is large and brief, requiring that an accurate measurement of Δ[Ca²⁺] be made with a low-affinity rapidly-responding indicator. Some low-affinity Ca²⁺ indicators monitor Δ[Ca²⁺] much more accurately than others, however, as reviewed here in measurements in frog twitch fibres with sixteen low-affinity indicators. This article also examines measurements and simulations of Δ[Ca²⁺] in mouse fast-twitch fibres. The simulations use a multi-compartment model of the sarcomere that takes into account Ca²⁺’s release from the SR, its diffusion and binding within the myoplasm, and its re-sequestration by the SR Ca²⁺ pump. The simulations are quantitatively consistent with the measurements and appear to provide a satisfactory picture of the underlying Ca²⁺ movements.     

A mechanism accounting for independence on starting length of tension increase in ramp stretches of active skeletal muscle at short half-sarcomere lengths

Based on previous experimental results of independence on starting length of the tension gradient in constant-velocity stretches of active skeletal muscle at muscle lengths including the ascending limb and the plateau of the tension-length relation, a possible physiological mechanism determining the tension increase in lengthening active muscle is discussed. Considering the sliding filament theory, it is suggested that the tension-length relation of a half-sarcomere in lengthening contractions is different from that in isometric contractions. The assumed mechanism predicts, among others, that the thick filament retains its shortened length in lengthening contractions starting from a half-sarcomere length where this filament is compressed. An example model is implemented and checked with simulations.     

Electromyographic basis of inaccurate movement; its dependence upon the mode of muscle contraction

The electromyographic basis of inaccurate performance was investigated in two rapid precision-grip skills controlled by concentric and eccentric muscle contractions respectively. Surface electromyograms, recorded from the first dorsal interosseous (DI), adductor pollicis (AP) and abductor pollicis brevis, were utilised to identify changes in the timing and intensity of muscle activation which may be responsible for inaccurate performance. The results showed that when fast precision-grip skills were controlled by concentric DI and AP muscle contraction, variations in the intensity of muscle contraction were responsible for inaccurate performance. However, when these skills were controlled by eccentric DI and AP muscle contractions, inaccurate performance resulted from variations in the timing of muscle activation. It was concluded that the nature of the deficiency in the patterns of muscle activation resulting in inaccurate performance was dependent upon the type of muscle contraction used in the skill.     

In situ NADH laser fluorimetry during muscle contraction in humans

The aim of the present study was to use nicotinamide adenine dinucleotide phosphate, reduced (NADH) fluorimetry, to investigate in situ NADH changes during muscle contraction in humans on an isokinetic dynamometer. Thirteen healthy male subjects each performed one maximal voluntary contraction (MVC) with the knee extensor muscle. The NADH muscle fluorescence was monitored by a double beam laser fluorimeter which uses an optical fibre, percutaneously inserted through a needle into the vastus lateral muscle, to guide the light. The NADH fluorescence was continuously measured at a wavelength of 337 nm. To estimate the haemodynamic artefact, blood backscattering was simultaneously determined at a wavelength of 586 nm. The fluorescence signal was recorded before, during and after contractions at 50% of MVC. The fibre was kept out of contact with the muscle during contractions at 100% of MVC and was only put into contact with it at the end of the contraction. At the onset of contractions at 50% of MVC, NADH fluorescence increased rapidly for 3 s and remained stable thereafter until exhaustion. After a muscle measurement had been made, the optical fibre was put successively into solutions of increasing NADH concentration to ascertain the relationship between the muscle fluorescence signal and the muscle NADH level. This procedure yielded estimated mean values for muscle NADH of 0.172 mmol.kg-1, SEM 0.028 and of 0.184 mmol.kg-1, SEM 0.027 after contractions at 50% and 100% of MVC, respectively, from a resting value of 0.087 mmol.kg-1, SEM 0.015. These results indicated that in situ laser fluorimetry could be used to evaluate NADH changes in humans during muscle contraction.(ABSTRACT TRUNCATED AT 250 WORDS)     

Predicting muscle force generation during fast-starts for the common carp Cyprinus carpio

Muscle contractile properties have been characterised for white myotomal muscle from the common carp Cyprinus carpio at 10, 15, and 20 °C. The time course of muscle force development was measured when one, two, or three stimuli were delivered at the onset of constant velocity shortening. As the shortening velocity increased several parameters decreased including the maximum force, the time course for the contraction and the relative duration of the deactivation compared to the activation. The maximum force and the relative rates of activation to deactivation for the contraction were relatively independent of temperature, whereas the duration of the contraction decreased with increasing temperature. A predictive model was developed which was based on fitting a modified Weibull distribution to these observations. The model was used to interpolate the expected contractile forces during cyclic length-changes. Measured and predicted values for force and power during such cyclic work-loop experiments showed an excellent agreement over the range of shortening regimes typically found during swimming behaviours. However, the predicted force was overestimated during the deactivation phase of the contractions when the shortening velocities exceeded those found during swimming. Accepted: 25 May 1999     

Gender modulates the energy cost of muscle contraction in untrained healthy subjects. A P magnetic resonance spectroscopy analysis

The forearm flexor muscles of 56 untrained volunteers (26 women and 30 men) were examined by ³¹P magnetic resonance spectroscopy, during a rest-exercise-recovery protocol, in order to document the impact of gender on muscle energetics. Absolute concentrations of high-energy phosphate compounds, intracellular pH and rates of aerobic and anaerobic ATP production were calculated. An inverse correlation was found between body mass index (BMI) and power output in women but not in men. After correcting for power output and BMI, the measured energy cost of contraction was twice larger for women than for men. This increase was also reflected in larger ATP production from aerobic and anaerobic pathways. This higher energy cost might be explained in part by differences in local muscle mass, a higher impact of fatness, but also by a reduced metabolic efficiency of muscle fibers in untrained women.     

The interrelation between aPKC and glucose uptake in the skeletal muscle during contraction and insulin stimulation

Contraction and insulin increase glucose uptake in skeletal muscle. While the insulin pathway, better characterized, requires activation of phosphoinositide 3‐kinase (PI3K) and atypical protein kinase (aPKC), muscle contraction seems to share insulin‐activated components to increase glucose uptake. This study aimed to investigate the interrelation between the pathway involved in glucose uptake evoked by insulin and muscle contraction. Isolated muscle of rats was treated with solvent (control), insulin, wortmannin (PI3K inhibitor) and the combination of insulin plus wortmannin. After treatment, muscles were electrically stimulated (contracted) or remained at rest. Glucose transporter 4 (GLUT4) localization, glucose uptake and phospho‐aPKC (aPKC activated form) were assessed. Muscle contraction and insulin increased glucose uptake in all conditions when compared with controls not stimulating an effect that was accompanied by an increase in GLUT4 and of phospho‐aPKC at the muscle membrane. Contracted muscles treated with insulin did not show additive effects on glucose uptake or aPKC activity compared with the response when these stimuli were applied alone. Inhibition of PI3K blocked insulin effect on glucose uptake and aPKC but not in the contractile response. Thus, muscle contraction seems to stimulate aPKC and glucose uptake independently of PI3K. Therefore, aPKC may be a convergence point and a rate limit step in the pathway by which, insulin and contraction, increase glucose uptake in skeletal muscle. Copyright © 2014 John Wiley & Sons, Ltd.     

Relation of NADH/NAD to contraction in vascular smooth muscle

The relationship of NADH/NAD to O2 consumption with respect to the different phases of contraction in vascular smooth muscle in response to a maximal depolarizing concentration of KCl was investigated. The NADH bound to cellular proteins could be distinguished from free NADH in whole tissue homogenates. Evidence suggested that the NADH was bound to pyruvate dehydrogenase and perhaps to other dehydrogenases since binding paralleled the changes in the activity of pyruvate dehydrogenase with contraction. The measured changes in NADH were attributed to that within the mitochondrial compartment since the contribution of reducing equivalents within the cytoplasmic compartment was negligible. During the phase of contraction in which force was initially being generated and at which O2 consumption was the highest, there was a net increase in NADH/NAD. After stable isometric force was maintained, at which time O2 consumption had returned to slightly above the basal pre-contraction level, there was a net decrease in NADH/NAD. Previous evidence indicates the phosphorylation potential (ATP/ADP) may decrease during this phase of contraction. It is concluded that contraction of vascular smooth muscle is accompanied by a changing pool of reducing equivalents. Factors which govern O2 consumption may change during the different phases of muscle contraction.     

Non-uniform mechanical activity of quadriceps muscle during fatigue by repeated maximal voluntary contraction in humans

To determine the non-uniform surface mechanical activity of human quadriceps muscle during fatiguing activity, surface mechanomyogram (MMG), or muscle sound, and surface electromyogram (EMG) were recorded from the rectus femoris (RF), vastus lateralis (VL), and vastus medialis (VM) muscles of seven subjects during unilateral isometric knee extension exercise. Time- and frequency-domain analyses of MMG and of EMG fatigued by 50 repeated maximal voluntary contractions (MVC) for 3 s, with 3-s relaxation in between, were compared among the muscles. The mean MVC force fell to 49.5 (SEM 2.0)% at the end of the repeated MVC. Integrated EMG decreased in a similar manner in each muscle head, but a marked non-uniformity was found for the decline in integrated MMG (iMMG). The fall in iMMG was most prominent for RF, followed by VM and VL. Moreover, the median frequency of MMG and the relative decrease in that of EMG in RF were significantly greater (P < 0.05) than those recorded for VL and VM. These results would suggest a divergence of mechanical activity within the quadriceps muscle during fatiguing activity by repeated MVC. Accepted: 19 January 1999     

Structural changes of the regulatory proteins bound to the thin filaments in skeletal muscle contraction by X-ray fiber diffraction

In order to clarify the structural changes related to the regulation mechanism in skeletal muscle contraction, the intensity changes of thin filament-based reflections were investigated by X-ray fiber diffraction. The time course and extent of intensity changes of the first to third order troponin (TN)-associated meridional reflections with a basic repeat of 38.4 nm were different for each of these reflections. The intensity of the first and second thin filament layer lines changed in a reciprocal manner both during initial activation and during the force generation process. The axial spacings of the TN-meridional reflections decreased by ∼0.1% upon activation relative to the relaxing state and increased by ∼0.24% in the force generation state, in line with that of the 2.7-nm reflection. Ca²⁺-binding to TN triggered the shortening and a change in the helical symmetry of the thin filaments. Modeling of the structural changes using the intensities of the thin filament-based reflections suggested that the conformation of the globular core domain of TN altered upon activation, undergoing additional conformational changes at the tension plateau. The tail domain of TN moved together with tropomyosin during contraction. The results indicate that the structural changes of regulatory proteins bound to the actin filaments occur in two steps, the first in response to the Ca²⁺-binding and the second induced by actomyosin interaction.     

Differences in the mechanism of collagen lattice contraction by myofibroblasts and smooth muscle cells

Both rat derived vascular smooth muscle cells (SMC) and human myofibroblasts contain α smooth muscle actin (SMA), but they utilize different mechanisms to contract populated collagen lattices (PCLs). The difference is in how the cells generate the force that contracts the lattices. Human dermal fibroblasts transform into myofibroblasts, expressing α‐SMA within stress fibers, when cultured in lattices that remain attached to the surface of a tissue culture dish. When attached lattices are populated with rat derived vascular SMC, the cells retain their vascular SMC phenotype. Comparing the contraction of attached PCLs when they are released from the culture dish on day 4 shows that lattices populated with rat vascular SMC contract less than those populated with human myofibroblast. PCL contraction was evaluated in the presence of vanadate and genistein, which modify protein tyrosine phosphorylation, and ML‐7 and Y‐27632, which modify myosin ATPase activity. Genistein and ML‐7 had no affect upon either myofibroblast or vascular SMC‐PCL contraction, demonstrating that neither protein tyrosine kinase nor myosin light chain kinase was involved. Vanadate inhibited myofibroblast‐PCL contraction, consistent with a role for protein tyrosine phosphatase activity with myofibroblast‐generated forces. Y‐27632 inhibited both SMC and myofibroblast PCL contraction, consistent with a central role of myosin light chain phosphatase. J. Cell. Biochem. 111: 362–369, 2010. © 2010 Wiley‐Liss, Inc.     

Cross bridge slippage induced by the ATP analogue AMP-PNP and stretch in glycerol-extracted fibrillar muscle fibres

Glycerol-extracted insect fibrillar muscle fibres in rigor exhibited both an elastic and a plastic phase in the length-tension diagram. The transition between these phases took place at a critical tension, the yield point or elastic limit. In the plastic phase the apparent static elastic modulus became zero, whereas the immediate elastic modulus (measured by rapid length changes completed within 4 ms) exhibited no abrupt change at the yield point. The tension value of the yield point (but not immediate stiffness) was lowered by addition of AMP-PNP and was partially restored by washing out AMP-PNP. The dependence of the critical tension at which plastic flow begins on cooperative cross bridge behaviour is discussed in terms of breaking and reforming acto-myosin linkages. Evidence is presented that addition of AMP-PNP induces slippage of cross bridges on the actin filament by affecting the interaction between myosin and actin.     

Caldesmon inhibits the rotation of smooth actin subdomain-1 and alters its mobility during the ATP hydrolysis cycle

Smooth muscle thin filaments have been reconstituted in muscle ghost fibers by incorporation of smooth muscle actin, tropomyosin and caldesmon. For the first time, rotation of subdomain-1 and changes of its mobility in IAEDANS-labeled actin during the ATP hydrolysis cycle simulated using nucleotides and non-hydrolysable ATP analogs have been demonstrated directly. Binding of caldesmon altered the mobility and inhibited the rotation of actin subdomain-1 during the transition from AM∗∗·ADP·Pi to AM state, resulting in inhibition of both strong and weak-binding intermediate states. These new results imply that regulation of actomyosin interaction by caldesmon during the ATPase cycle is fulfilled via the inhibition of actin subdomain-1 rotation toward the periphery of the thin filament, which decreases the area of the specific binding between actin and myosin molecules and is likely to underlie at least in part the mechanism of caldesmon-induced contractility suppression.