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PubMedDate not supplied

Intensification of the 5.9-nm actin layer line in contracting muscle.

According to the cross-bridge model of muscle contraction, an interaction of myosin heads with interdigitating actin filaments produces tension. Although X-ray equatorial diffraction patterns of active (contracting) muscle show that the heads are in the vicinity of the actin filaments, structural proof of actual attachment of heads to actin during contraction has been elusive. We show here that during contraction of frog skeletal muscle, the 5.9-nm layer line arising from the genetic helix of actin is intensified by as much as 56% of the change which occurs when muscle enters rigor, using a two-dimensional X-ray detector. This provides strong structural evidence that myosin heads do in fact attach during contraction.

Actins↗
PubMed1979

Pentobarbitone and skeletal muscle contractions: on the interaction with the effect elicited by the beta-adrenoceptor agonist, terbutaline.

The soleus, a slow-contracting muscle, and the extensor digitorum longus (EDL), a fast-contracting muscle from guinea-pig were prepared for isometric recording in vitro. Subtetanic contractions were evoked by transmural field-stimulation. Pentobarbitone increased the force of contraction in both muscles. In the soleus it shifted the stimulation frequency-response curve to the left. Terbutaline caused a decrease in the force of subtetanic contractions of the soleus, an effect which was dependent on the stimulation frequency. In the presence of pentobarbitone, the stimulation frequency had to be lowered by about 2 HZ in order to maintain the optimum response to terbutaline. The EDL responded to terbutaline with an increased force of contraction. In this case the stimulation frequency was less critical and the effects were the same in the presence and in the absence of pentobarbitone. Experiments with alpha-chloralose yielded results similar to those obtained with pentobarbitone.

Animals↗
PubMedDate not supplied

Actomyosin structure in contracting muscle detected by rapid freezing.

It is now widely accepted that the ATP-induced active sliding of adjacent thin and thick filaments mediated by myosin heads (cross-bridges) is responsible for muscle contraction. Despite intensive studies, the behaviour of the myosin heads during muscle contraction is still unclear. Recent progress in the rapid freezing electron microscope technique has greatly improved the temporal resolution of the images that can be obtained. Here, we report a new type of actomyosin structure captured by rapid freezing. We have analysed images from thin sections of freeze-substituted rabbit skeletal muscle rapidly frozen during isometric contraction. For comparison, we also studied relaxed and rigor muscles. Our results show that, during isometric contraction, most myosin heads are regularly arrayed along the helix of the actin filaments and that this actomyosin structure appears to be distinct from that observed in rigor muscle.

Actins↗
PubMed1988

A self-induced translation model of myosin head motion in contracting muscle. I. Force-velocity relation and energy liberation.

In our previous model, it was assumed that the two heads of myosin act co-operatively in producing force for the sliding of actin filaments relative to myosin filaments. We eliminate the assumption of co-operativity in the present model, following the conclusion by Harada and co-workers that a co-operative interaction between the two heads of myosin is not essential in producing actin filament movement. We assume that (1) a myosin head activated by ATP hydrolysis binds to the thin filament at a definite angle and does not do the power stroke, i.e. does not change its orientation during attachment, (2) a potential of force acting on the myosin head is induced around the thin filament when an ATP-activated myosin head binds to an actin molecule in the thin filament, and (3) the potential remains for a while after detachment of the myosin head and statistically controls the direction of thermal motion of the myosin head, so that the myosin head translates toward the Z-line as a statistical average. We did calculations on these assumptions with a mean tension approximation and got the following results. (a) The calculated force-velocity relation in muscle contraction is in fairly good agreement with experimental observation, including the give phenomenon that lengthening velocity becomes very large for a force about twice the isometric tension. (b) The calculated rate of energy liberation during muscle contraction as a function of load on muscle is in good agreement with experimental results. (c) The calculated distance over which a myosin molecule moves along the thin filament during one ATP hydrolysis can be more than 60 nm under unloaded conditions.

Energy Metabolism↗
PubMed1994

Tension development and duty cycle affect Qpeak and VO2peak in contracting muscle.

Ten canine gastrocnemius-plantaris muscle preparations were stimulated in situ to determine the interaction between tension development and the duty cycle in determining Qpeak and VO2peak. The muscle was stimulated with supramaximal voltage using four different stimulation protocols: 1) 5 twitches.s-1 (Tw), 2) 1 train.s-1-200 ms (1-200), 3) 1 train.s-1-300 ms (1-300), and 4) 2 trains.s-1-100 ms (2-100). Arterial and venous blood were sampled and Qpeak measured for determination of VO2peak. The total tension developed per second was integrated and averaged over 1 s (TDa) and used as an index of work of the muscle for each condition. The Qpeak and VO2peak were greater (P < 0.05) in the 1-200 condition compared to all other conditions. Further, Qpeak and VO2peak were greater (P < 0.05) in both the 1-300 and 2-100 conditions than during Tw: Qpeak (ml.kg-1.min-1) (mean +/- SE) for (Tw) = 928 +/- 65; (1-200) = 1368 +/- 102; (1-300) = 1150 +/- 96; (2-100) = 1189 +/- 89; VO2peak (ml.kg-1.min-1) for (Tw) = 108 +/- 8; (1-200) = 159 +/- 9; (1-300) = 135 +/- 11; (2-100) = 137 +/- 8. The TDa was significantly different among all conditions: TDa (N.kg-1) for (Tw) = 443 +/- 56; (1-200) = 606 +/- 81; (1-300) = 722 +/- 79; (2-100) = 522 +/- 41. We interpret these findings as an indication that the interaction of the duty cycle and tension development is a prime determinant of blood flow during muscle contractions.

Animals↗
PubMed2002

Effect of rhythmic tetanic skeletal muscle contractions on peak muscle perfusion.

The purpose of this investigation was to examine the effect of rhythmic tetanic skeletal muscle contractions on peak muscle perfusion by using spontaneously perfused canine gastrocnemii in situ. Simultaneous pulsatile blood pressures were measured by means of transducers placed in the popliteal artery and vein, and pulsatile flow was measured with a flow-through-type transit-time ultrasound probe placed in the venous return line. Two series of experiments were performed. In series 1, maximal vasodilation of the muscles' vascular beds was elicited by infusing a normal saline solution containing adenosine (29.3 mg/min) and sodium nitroprusside (180 microg/min) for 15 s and then simultaneously occluding both the popliteal artery and vein for 5 min. The release of occlusion initiated a maximal hyperemic response, during which time four tetanic contractions were induced with supramaximal voltage (6-8 V, 0.2-ms stimuli for 200-ms duration at 50 Hz, 1/s). In series 2, the muscles were stimulated for 3 min before the muscle contractions were stopped for a period of 3 s; stimulation was then resumed. The results of series 1 indicate that, although contractions lowered venous pressure, muscle blood flow was significantly reduced from 2,056 +/- 246 to 1,738 +/- 225 ml x kg(-1) x min(-1) when contractions were initiated and then increased significantly to 1,925 +/- 225 ml x kg(-1) x min(-1) during the first 5 s after contractions were stopped. In series 2, blood flow after 3 min of contractions averaged 1,454 +/- 149 ml x kg(-1) x min(-1). Stopping the contractions for 3 s caused blood flow to increase significantly to 1,874 +/- 172 ml x kg(-1) x min(-1); blood flow declined significantly to 1,458 +/- 139 ml x kg(-1) x min(-1) when contractions were resumed. We conclude that the mechanical action of rhythmic, synchronous, maximal isometric tetanic skeletal muscle contractions inhibits peak muscle perfusion during maximal and near-maximal vasodilation of the muscle's vascular bed. This argues against a primary role for the muscle pump in achieving peak skeletal muscle blood flow.

Adenosine↗
PubMed1992

[Control of isometric muscle contraction in muscle hypotonia of central origin: EMG mapping analysis].

Electromyographic and mechanographic investigations in patients with muscular hypotonia, which is, for instance, a side-effect after stereotactic treatment of tremor syndromes, permit the presumption that in this sensomotor open-loop situation the decreased muscular resistance to stretching during isometric contraction (initial stiffness) is caused by changes of muscular innervation pattern. Probably, the innervation pattern during tonic activity is changed by a shift of a more tonic motoneurone behaviour to motoneurone activities with predominantly phasic characteristics. In 17 controls and 4 patients with muscular hypotonia caused by stereotactic lesions of VIM area (treatment of tremor syndromes) the EMG of right and left side forearm flexors (especially the activity of the M. biceps brachii) was investigated by a sophisticated, topographically oriented 16-channel-surface-EMG-technique ("EMG-Mapping") during slight isometric contraction. EMG-Maps of forearm flexors (especially of M. biceps brachii) in patients with centrally evoked muscular hypotonia demonstrate that in these open-loop conditions the motor control is changed. For this the reason could be a shift of the activated motor units from a predominantly static to a more phasic functional behaviour. The latest results on muscular activation processes in cats support this presumption.

Adult↗
PubMed1999

Effects of muscle contraction on skeletal muscle blood flow: when is there a muscle pump?

PURPOSE: The purpose of this study was to determine the effects of rhythmic muscle contraction on the dynamics of venous outflow in rat skeletal muscle. METHODS: The effects of frequency and duration of tetanic contraction on venous blood flow (BF) were examined with transonic flow probes placed on the femoral artery and vein. RESULTS: Results reveal that instrumentation of the venous system with cannulas or flow probes alters vascular mechanics so that the muscle pump effect is masked. Measurements conducted without instrumentation of the venous vasculature in situ, as well as experiments with conscious exercising animals, indicate that the muscle pump enhances BF during exercise. Also, recent in vivo studies of humans indicate an important role for the muscle pump. In contrast, results reported herein and recent results from in situ experiments, which allow control of more parameters, indicate that there is no measurable muscle pump effect on BF during rhythmic muscle contraction. Review of the literature indicates that many in vitro/in situ experiments used instrumented veins that may have altered venous vascular mechanics and the interactions of muscle contraction and venous vascular mechanics, thus minimizing or abolishing the muscle pump effect. CONCLUSIONS: The muscle pump contributes to the initial increase in BF at exercise onset and to maintenance of BF during exercise.

Animals↗
PubMed2003

From sarcomere to cell: an efficient algorithm for linking mathematical models of muscle contraction.

Two classes of mathematical framework have previously been developed to model active tension generation in contracting muscle. Cross-bridge models of muscle are biophysically based but computationally expensive to solve, and thus unsuitable for embedding in spatially distributed continuum representations. Fading memory models are computationally efficient but provide limited biophysical insight. In this study a novel computational method is proposed for coupling these two frameworks such that biophysical events can be determined and computational tractability maintained. Within the cross-bridge model, the functional forms of the distribution of cross-bridges, as a function of strain in each state, are approximated using the distribution moment approach. Using the variables of area, mean and standard deviation of each distribution, analytic expressions are developed to calculate the temporal dynamics of stiffness, tension and energy. A root finding method is employed to adjust the variables such that the temporal dynamics of the cross-bridge model match those of an equivalent fading memory model. The method is demonstrated for sinusoidal perturbations in length at two frequencies, with an approximate 30-fold increase in computational efficiency over a conventional technique for finding a solution to the cross-bridge model.

Adenosine Triphosphate↗
PubMed1999

Regulation of Limulus skeletal muscle contraction.

Skeletal muscle contraction of Limulus polyphemus, the horseshoe crab, seemed to be regulated in a dual manner, namely Ca2+ binding to the troponin complex as well phosphorylation of the myosin light chains (MLC) by a Ca2+/calmodulin-dependent myosin light chain kinase. We investigated muscle contraction in Limulus skinned fibers in the presence of Ca2+ and of Ca2+/calmodulin to find out which of the two mechanisms prevails in Limulus skeletal muscle contraction. Although skinned fibers revealed high basal MLC mono- and biphosphorylation levels (0.48 mol phosphate/mol 31 kDa MLC; 0.52 mol phosphate/mol 21 kDa MLC), the muscle fibers were fully relaxed at pCa 8. Upon C2+ or Ca2+/calmodulin activation, the fibers developed force (357+/-78.7 mN/mm2; 338+/-69.7 mN/mm2, respectively) while the MLC phosphorylation remained essentially unchanged. We conclude that Ca2+ activation is the dominant regulatory mechanism in Limulus skeletal muscle contraction.

Animals↗
PubMed1980

On the possible role of potassium ions in the action of terbutaline on skeletal muscle contractions.

The soleus, a slow-contracting, and the extensor digitorum longus (EDL), a fast-contracting muscle from guinea-pig were prepared for isometric recording in vitro, and the effect of terbutaline, a beta-adrenoceptor agonist, was studied during various experimental conditions. In the soleus terbutaline caused an initial depression of the force of contraction followed by an increase. The degree of fusion of subtetanic contractions was reduced throughout the experiment. The EDL responded with an increased force only. The force of contraction of the two muscles increased with the K+-ion concentration of the medium up to 6 mM. Higher concentrations of K+ caused a depression which was partly prevented by terbutaline. When the contractions of the soleus muscle had been depressed by excess K+, depletion of K+ or by ouabain, terbutaline restored the twitch tension. It is suggested that the contractile machinery of the muscles is controlled by the ionic balance which in turn is changed following beta-adrenoceptor stimulation.

Animals↗
PubMed1998

Asymmetrical hemispheric activation and behavioral persistence: effects of unilateral muscle contractions.

Contractions of the left hand and of the left side of the lower third of the face induce negative emotional states whereas right-sided contractions induce positive states. Contractions also have mood-congruent influences on perception. This article reports that contractions affect behavior as well. Persistence in attempting to solve insoluble problems is greater following right-sided contractions than following left-sided contractions. This effect is unrelated to dominance of the contracted muscles because right-handed individuals tend to be left-face dominant. Results support the hypothesis that unilateral contractions activate the contralateral cerebral hemispheres and arouse the hemispheres' respective functions with regard to emotion and behavior.

Adult↗
PubMed1994

A model of the release of myosin heads from actin in rapidly contracting muscle fibers.

We describe a model that relates the maximum shortening velocity of a muscle fiber, Vm, to the kinetics of the dissociation of a myosin head from actin. At Vm, the positive work exerted by cross-bridges attached in the powerstroke must be balanced by cross-bridges that have been carried by movement of the filaments into a region where they exert a negative force. This balance allows one to relate Vm and the rate of cross-bridge detachment. Studies of actomyosin kinetics suggest that at high substrate, detachment should be limited by a slow protein isomerization (approximately 50 s-1) that precedes ADP release. This rate is too slow to be easily accommodated in existing models. However, a slow rate for cross-bridge dissociation, similar to that of the isomerization, is predicted if previous models are modified to include rapid detachment of cross-bridges that have been carried so far into the negative force region that their free energy exceeds that of the detached state. The model also explains another aspect of muscle contraction: at high shortening velocities, the observed rate of ATP hydrolysis is low, because a cross-bridge can interact with multiple actin binding sites before releasing the hydrolysis products and binding another ATP.

Actins↗
PubMed1992

Rapid regeneration of the actin-myosin power stroke in contracting muscle.

At the molecular level, muscle contraction is the result of cyclic interaction between myosin crossbridges, which extend from the thick filament, and the thin filament, which consists mainly of actin. The energy for work done by a single crossbridge during a cycle of attachment, generation of force, shortening and detachment is believed to be coupled to the hydrolysis of one molecule of ATP. The distance the actin filament slides relative to the myosin filament in one crossbridge cycle has been estimated as 12 nm by step-length perturbation studies on single fibres from frog muscle. The 'mechanical' power stroke of the attached crossbridge can therefore be defined as 12-nm shortening with a force profile like that shown by the quick recovery of force following a length perturbation. According to this definition, power strokes cannot be repeated faster than the overall ATPase rate. Here, however, we show that the power stroke can be regenerated much faster than expected from the ATPase rate. This contradiction can be resolved if, in the shortening muscle, the free energy of ATP hydrolysis is used in several actin-myosin interactions consisting of elementary power strokes each of 5-10 nm.

Actins↗
PubMed1969

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↗
PubMed2002

Coordination of the two heads of myosin during muscle contraction.

We have used luminescence resonance energy transfer between regulatory light chains (RLC) to detect structural changes within the dimeric myosin molecule in contracting muscle fibers. Fully functional scallop muscle fibers were prepared such that each myosin molecule contained a terbium-labeled (luminescent donor) RLC on one head and a rhodamine-labeled (acceptor) RLC on the other. Time-resolved luminescence energy transfer between the two heads increased upon the transition from relaxation (ATP) to contraction (ATP plus Ca) and increased further in rigor (no ATP). Combined with experiments on mutant RLCs labeled specifically at other sites, these results support a model in which the force-generating weak-to-strong transition causes one myosin LC domain to tilt through a 30 degrees angle toward the other, thus acting as a coordinated lever arm.

Animals↗
PubMed2006

X-ray interference studies of crossbridge action in muscle contraction: evidence from quick releases.

We have used a high-resolution small angle X-ray scattering system, together with a high-performance CCD camera, on the BioCAT beamline at the APS synchrotron radiation facility at the Argonne National Laboratory, to study X-ray interference effects in the meridional reflections generated by the arrays of myosin crossbridges in contracting muscle. These give information about axial movements of the myosin heads during contraction with sub-nanometer resolution. Using whole intact muscle preparations (frog sartorius) we have been able to record the detailed behavior of M3 (the first order meridional reflection from the myosin crossbridges, at 14.56 nm) at each of a number of quick releases of increasing magnitude, on the same specimen, and at the same time make similar measurements on higher order myosin meridional reflections, particularly M6. The latter provides information about the dispersion of lever arm angles of the actin-attached myosin heads. The observations show that in isometric contraction the lever arm angles are dispersed through +/- 20-25 degrees on either side of a mean orientation that is about 60 degrees away from their orientation at the end of the working stroke: and that they move towards that orientation in synchronized fashion, with constant dispersion, during quick releases. The relationship between the shift in the interference fringes (which measures the shift of the myosin heads scattering mass towards the center of the sarcomere, and the changes in the total intensity of the reflections, which measures the changes in the axial profile of the heads, is consistent with the tilting lever arm mechanism of muscle contraction. Significant fixed contributions to the meridional reflections come from unattached myosin heads and from backbone components of the myosin filaments, and the interaction of these with the contributions from actin-attached myosin heads determines the behavior of these reflections.

Animals↗