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Cross-bridge cooperativity during isometric contraction and unloaded shortening of skeletal muscle.

Whether the two heads of skeletal muscle myosin work independently or cooperatively remains an open question in muscle biophysics. While individual myosin heads are sufficient for ATPase activity (Reisler (1980) J Mol Biol 138: 93-107) and force production (Harada et al. (1987) Nature 326: 805-808), it has also been reported that in situ, the two heads of a myosin molecule work cooperatively (Chaen et al. (1986) J Biol Chem 261(29): 13,632-13,636). To examine the role of cross-bridge cooperativity on isometric contraction and unloaded shortening we progressively inactivated myosin cross-bridges via titration with para-phenylenedimaleimide. The resting fiber ATPase was measured to provide an estimate of the fraction of active cross-bridges remaining during the titration. Isometric force and unloaded shortening velocity decline more rapidly than the resting ATPase as the titration proceeds. This is inconsistent with models for independent force generation and suggests cooperative action of myosin cross-bridges when muscle is isometrically contracting or shortening under zero load. However the degree of cooperativity depends on the type of muscle activity. While isometric force declines in a manner consistent with pair-wise cooperative action of myosin heads, unloaded shortening velocity declines more rapidly (greater cooperativity). Therefore, myosin cross-bridges in situ may be capable of at least two types of cooperative interactions, pair-wise cooperativity (when isometric) and another form of cooperativity that is sensitive to longer range interactions transmitted from other cross-bridges in the ensemble (during unloaded shortening).

Adenosine Triphosphatases↗

The effects of local anaesthetics on the isometric contraction of the isolated hemidiaphragm of the rat.

Local anaesthetics (benzocaine, tetracacine, lidocaine, cocaine, procaine) were found to depress both Td and dT/dt max of the isometric contraction of the isolated hemidiaphragm of the rat during direct electrical stimulation. The same substances were found to antagonize the action of aminophylline on Td and dT/dt max of the isolated hemidiaphragm during direct stimulation. The effect of higher concentrations of aminophylline was even reversed by cocaine, procaine and lidocaine. Increasing of calcium concentrations did not reverse the antagonistic action of aminophylline on Td and dT/dt max of the isometric contraction of the isolated hemidiaphragm. It is suggested that a change in the muscle membrane is responsible for the observed action of local anaesthetics on Td and dT/dt max, as well as for their antagonistic action towards aminophylline.

Aminophylline↗

Distribution of muscle fiber conduction velocity of M. biceps brachii during voluntary isometric contraction with use of surface array electrodes.

Surface electromyogram (EMG) was recorded in m. biceps brachii during the contractions of 20, 40 and 60% of maximum voluntary isometric contraction (MVC) in twelve healthy male subjects, using surface array electrodes. The distribution of muscle fiber conduction velocity (MFCV) was found directly using the averaging technique and the cross-correlation function technique. MFCVs in the region of 20-40 mm measured from end-plate denoted constant value of about 4 m/s in 20% MVC, while MFCVs in the region around end-plate and tendons showed about 10 m/s in 20% MVC. The values of MFCV depended on the contraction levels of muscle. The model for the generation of MFCV which considered the ensemble of muscle fibers with the shape of a cone was proposed. The theoretical values of MFCV by the muscle fiber ensemble model (MFE model) proposed in the paper showed in good agreement with the experimental results.

Action Potentials↗

Organizing principles for single-joint movements. IV. Implications for isometric contractions.

1. Normal human subjects made isometric pulse and step contractions about the elbow to visually defined target torques of different amplitudes and at different rates. We measured joint torque and electromyograms (EMG) from two agonist and two antagonist muscles. 2. When the task specification requires that the subject explicitly alter the rate at which torque is increased, the rates of rise of the agonist and antagonist EMG bursts covary with the rate of rise of the torque. For pulses of torque the duration of motoneuron excitation varies with the duration of the task-defined contractile event. 3. When a subject is asked to generate torques of different amplitudes without specifying a time interval, torque amplitude is positively correlated with how long, and therefore how high, the EMG rose. Subjects usually proportionately covary the strength of the agonist and antagonist contractions but are not constrained to do so. Some subjects use a strategy of varying the antagonist inversely with the agonist contraction. 4. We extend the organizing principles for the control of movement about a single joint to the control of isometric torque. These rules state that control of torque about a single joint is exercised by one of two strategies: the speed-sensitive strategy modulates the rate at which contraction rises by varying the intensity of motoneuron-pool excitation. The speed-insensitive strategy varies the duration over which contraction rises but does not change the rate. These two respective patterns of torque emerge from pulse-height and pulse-width modulation of motoneuron-pool excitation. 5. The rules defining speed-sensitive and speed-insensitive strategies for movements are broadened for isometric contractions because of the wider range of torque patterns that we observe under these conditions. We propose a step-excitation component for prolonged isometric step contractions and slowly rising ramp patterns of excitation for contractions that develop over several hundreds of milliseconds. 6. The choice of strategies is based on task-specific torque requirements. The same two strategies that control torque to produce movement apply to the control of isometric torque. Unlike movements, however, isometric tasks are more often controlled by a blending of the two patterns. Possible reasons for this are discussed.

Electromyography↗

Relation between intrinsic viscoelasticity and activation level of the human finger muscle during voluntary isometric contraction.

The purpose of the present study was to isolate the length perturbation-evoked force attributed to intrinsic muscle viscoelasticity, and to investigate the relation between muscle viscoelasticity and the level of muscle activation during isometric contraction in five healthy male subjects. A small length perturbation (stretching or contraction) was applied to the flexor pollicis longus muscle while the subject maintained constant isometric force; the time courses of the length perturbation was found to be almost identical in all the experiments. The force (Fv) induced by the muscle viscoelasticity was calculated using the equation Fv = F - Fc - Fp over an interval of 35 ms after the onset of perturbation, where F is the measured force, Fc is the tonic isometric force before the onset of perturbation and Fp is the force at rest obtained from the same length perturbation. The force response attributed to the stretch reflex is not included during this interval. These experiments were repeated at varying levels of isometric force. An almost linear relationship was obtained between the muscle viscoelasticity-induced force and the tonic isometric force during both the stretching and contraction of the muscle, i.e. the intrinsic muscle viscoelasticity varied almost linearly with the level of isometric contraction.

Adult↗

The control of blood flow through human forearm muscles following brief isometric contractions.

1. The blood flow through the forearm was measured 2 sec after single, brief isometric hand-grip contractions. The tension and duration of those contractions varied from 10 to 100% of the maximal voluntary contraction (m.v.c.) and from 2 to 12 sec, respectively. 2. The blood flow increased linearly with tension up to about 60% m.v.c. but further increases in tension, up to 100% m.v.c., did not elicit higher blood flows than were found at 60% m.v.c. The same relationship between tension and the resultant blood flow held for all durations of contractions, from 2 to 12 sec. The blood flow immediately after (2 sec) contractions at a given tension increased linearly with the duration of the contraction, from 2 to 12 sec. Maximal exercise blood flow was approached only in response to the longest contractions (12 sec) at tensions of 60% m.v.c. or higher. 3. Brief alterations (2--5 sec) of transmural pressure across blood vessels did not result in a significant change of blood flow, either in the resting forearm or when the vessels were dilated by brief, isometric contractions. When the tension was applied or released either rapidly or gradually ('ramp' contractions) there was no correlation between the rate of change of stretch on arterial vessels and the resultant blood flow. However, there was a direct relationship between a force--time integral (duration of contraction x peak tension) and blood flow. All these results make it clear that changes in blood flow in the forearm elicited by brief isometric contractions are not the result of a myogenic reflex but are metabolically induced. 4. Successive contractions exerted at 60% m.v.c. for 4 sec induced a blood flow of 21.2 +/- 1.6 ml.min-1.100 ml.-1 when a rest interval of 8 sec was allowed between the contractions. Blood flows remained constant at this submaximal level, even when muscular fatigue was induced, and when there was an accompanying large increase in blood pressure. 5. Isometric muscular activity by the contralateral arm which resulted in fatigue, associated with a large increase in mean blood pressure, did not alter the level of vasodilation that was induced by brief, isometric contractions in the 'test' arm. 6. It is suggested that the vasodilatation in response to intermittent isometric contractions is the result of metabolic vasodilatation of distal segments and continued sympathetic vasoconstriction of the proximal segments of the forearm vascular bed.

Blood Pressure↗

Frequency of acoustic myography during isometric contraction of fresh and fatigued muscle and during dynamic contractions.

The frequency of the acoustic myographic (AMG) signal was examined during fresh and fatigued isometric contractions of quadriceps and during dynamic contractions of biceps brachii (BB) in healthy subjects. Recordings were obtained from quadriceps over a range of forces between 10% and 100% maximal voluntary contraction prior to, and 15 minutes after, a fatiguing exercise. Recordings from BB were obtained over a range of submaximal forces (0-8.5 kg) during concentric and eccentric contractions. The mean power frequency (MPF) of the AMG signal was analyzed during each of these contractions by fast-Fourier transform (FFT). The MPF was not significantly different (P > 0.05) during fresh and fatigued contractions of quadriceps and increased quadratically with force in both states (r = 0.81, fresh; r = 0.77, fatigued). During concentric contractions of BB the MPF initially increased with force, but then decreased at the heavier loads (> 5.5 kg). The MPF of eccentric contractions did not significantly (P > 0.05) alter with force. The AMG MPF was within a similar low frequency range for both muscles, during different types of contraction, and was unaltered with fatigue.

Adolescent↗

Myofibrillar ATPase activity during isometric contraction and isomyosin composition in rat single skinned muscle fibres.

1. Myofibrillar ATPase activity, isometric tension (Po) and unloaded shortening velocity (Vo) were determined in single skinned fibres isolated from rat hindlimb muscles during maximal calcium activation at 12 degrees C. In each fibre, myosin heavy chain (MHC) isoforms were identified using electrophoresis and immunocytochemistry. ATPase activity was determined spectrophotometrically from NADH oxidation in a coupled enzyme assay. 2. On the basis of their MHC isoform composition, the fibres (n = 102) were divided into five groups containing the slow isoform, I MHC, or one of the fast isoforms, IIB MHC, IIA MHC, IIX MHC, or a mixture of the latter three. ATPase activity was significantly higher in IIB than in 2X and IIA fibres (0.230 +/- 0.010, 0.178 +/- 0.023 and 0.168 +/- 0.026 nmol mm-3 s-1, respectively). Mixed fibres had intermediate values. ATPase activity in slow fibres was considerably less (0.045 +/- 0.006 nmol mm-3 s-1). 3. The ratio between ATPase activity and Po, i.e. tension cost, was found to be 2.90 +/- 0.09, 2.56 +/- 0.14, 1.89 +/- 0.22, 1.52 +/- 0.13 and 0.66 +/- 0.004 pmol ATP nM-1 mm-1 s-1 in IIB, mixed, IIX, IIA and slow fibres, respectively. All the differences were statistically significant except that between IIA and IIX fibres. 4. Within each group of fibres with the same MHC composition, ATPase activity was found to correlate with Po, but not Vo. However, ATPase activity was found to correlate with Vo when all the fibre types were pooled together. 5. In thirty-seven fast fibres the MLC ratio, i.e. the proportion of the fast alkali light chain isoform, MLC3f, to the amount of the regulatory light chain, MLC2f, was determined. IIB fibres had the highest proportion of MLC3f and IIA fibres, the lowest. 6. A multiple regression analysis, used to distinguish between the effects of MHC and MLC composition, showed that ATPase activity was insensitive to the MLC ratio, whereas it had a significant impact on Vo. 7. The results obtained in this study indicate that in rat skeletal muscle fibres: (a) ATPase activity during isometric contractions and tension cost are strongly dependent on MHC isoform composition, and (b) there is no evidence that the alkali MLC ratio is a determinant of ATPase activity.

Adenosine Triphosphate↗

A simple model of force generation by skeletal muscle during dynamic isometric contractions.

The force that an isometric skeletal muscle will produce in response to time-varying stimulation ("dynamic isometric" force) is important both for understanding muscle function and for designing neuroprostheses. This paper reports a model for predicting the force produced by an isometric skeletal muscle at rest length in response to a wide range of stimulation patterns. The model consists of two linear, first-order systems separated by a static nonlinearity. The rate constant of the second first-order system varies with force level. The model was validated using three cat soleus and three cat plantaris muscles. The following whole-nerve stimulation trains were used: single pulses (twitches), 2-4 pulses, constant rates, triangularly modulated interpulse intervals, and randomly modulated interpulse intervals. The model reproduced most responses accurately. The model shows that a force-dependent rate constant is essential for model validity, and could be used in the control of neuroprostheses.

Analog-Digital Conversion↗

Systolic blood pressure responses during isometric contractions of large and small muscle groups.

The purpose of this study was to test for a difference between the systolic blood pressure responses to voluntary contractions of large and small muscle groups of the upper extremity. Systolic blood pressure was measured at approximately 20 sec intervals during sustained isometric contractions of the index finger adductors and handgrip muscles contracting at 40% of maximal voluntary contraction (MVC). Contractions were terminated when EMG activity exceeded 10% of maximal activity of selected accessory muscles or when the 40% MVC tension could no longer be held constant (+/-10%). The slopes of the regression lines representing systolic blood pressure responses to sustained isometric contraction of a small and large muscle group of the upper extremity were significantly different (p less than .001). Older theories that pressor responses are determined only by the % MVC and not by the mass of contracting muscle need to be reexamined.

Adult↗

Electrical and mechanical changes in human soleus muscle during sustained maximum isometric contractions.

Experiments were designed to evaluate changes in the electrical activation and force generating capabilities of human soleus muscle during sustained, maximum isometric contractions. Eighteen experiments were conducted on 7 healthy subjects. Surface EMG, and in select cases, intramuscular fine wire EMG recordings, were made to assess the electrical activation of soleus. Subjects performed maximum isometric plantarflexion contractions of 1-3 min during which time supramaximal electrical pulses were delivered to the tibial nerve at 5-s intervals to elicit maximum M waves. M wave areas were assessed for evidence of neuromuscular junction failure. The results revealed that, on average, maximum force declined to 80% of unfatigued maximum by 60 s of effort, 74% by 90 and 120 s, and 70% by 180 s. M waves were stable for efforts up to 3 min, thereby providing little evidence for neuromuscular junction failure. In 3 experiments, total spike counts from intramuscular recordings displayed a 50% reduction in firing by 30 s of effort, with little additional slowing for up to 3 min. Although all of the fatigue-induced electrical and mechanical alterations in muscle activation reported earlier for intrinsic hand and foot muscles were verified in these experiments on soleus, the magnitudes and time courses of these changes were quite different. All changes were consistent with a muscle designed to optimally resist fatigue.

Action Potentials↗

Isometric contraction of the abductor digiti minimi muscle in man.

Isometric contraction of the abductor digiti minimi muscle (ADM) has been studied in six normal subjects. Twitch contraction times of ADM ranged from 60 to 68 ms and twitch torque ranged from 2·33 to 6·24 × 10(-3) Nm. In three subjects torque declined by an average of 31% after tetanization at 50 Hz for 30 seconds but there was no similar diminution in the evoked muscle action potential suggesting that the fatigue arose from intrinsic muscular mechanisms. A marked decline in tetanic torque occurred with continued tetanization in two subjects for a total of five minutes, but this change was accompanied by a decrease in the muscle action potential. In six subjects threshold stimulation to the ulnar nerve at the wrist and to various sites over the motor point of ADM allowed 55 threshold twitch contractions to be identified after averaging. A unimodal range of contraction times ranging from 40-100 ms was found and this was confirmed by additional experiments in two subjects in whom 30 threshold twitch contractions were identified using a needle electrode to stimulate various sites in the motor point. Tetanization at 50 Hz was performed using threshold stimulus levels. Nine threshold tetanic contractions were evoked in two subjects. In eight tetanic torque progressively fatigued to between 14 and 20% within 60-90 seconds, but, in one tetanic contraction, torque proved relatively fatigue resistant. These results suggest that there is a homogeneous group of motor units in ADM (with respect to contraction time) and that this group contains what are probable fast twitch fatigue sensitive and fatigue resistant motor units. No evidence of a distinct group of slow twitch units was found.

Adult↗

Time-course of force production by fast isometric contraction of the knee extensor in young and elderly subjects.

Rapid force production by isometric contraction of the knee extensor was examined in a wide range of force output for 12 healthy elderly (65-86 years) and 12 young (20-35 years) subjects. Time-course of tension development and duration of first burst EMG activities (AG-1) of the vastus medialis muscle were compared between both groups. Significant increase in the elderly as compared with the young was found in duration from the onset of the EMG to the rise of tension (TLT), but not in time from the rise to peak of force (FTmax), although FTmax tended to be longer in the elderly than the young at three different levels of force output. AG-1 duration was also prolonged in the elderly but the difference was not significant. The prolongation of TLT in the elderly suggests that spatio-temporal recruitments of the motor units and/or the percentage of fast twitch fibers decreases with aging.

Adult↗

X-ray evidence for the elongation of thin and thick filaments during isometric contraction of a molluscan smooth muscle.

The elongation of thin and thick filaments during isometric contraction of a molluscan smooth muscle was studied by measuring spacing changes of meridional reflections in the medium-angle X-ray diffraction pattern. X-ray patterns from the anterior byssus retractor muscle of Mytilus edulis in the resting, active, and catch states were taken from the same part of a muscle bundle at a fixed specimen-to-detector distance, using imaging plates and 10 s exposure to synchrotron radiation. The third-order reflection (9.2 A) of the axial period of actin, and the fourteenth-order reflection (10.4 A) of the axial subunit-repeat of the thick filament are increased in spacing in the active and catch states. From accurately measured changes in the axial distance of the 9.2 A layer line from the origin, thin filament elongations in the active and catch states are estimated to be 0.48 and 0.32%, respectively, in a muscle that maintains a tension of 12.2 kg cm-2 in the active state and 9.8 kg cm-2 in the catch state. Thick filament elongations in the active and catch states are similarly estimated to be 0.33, and 0.28%, respectively, based on the axial shift of the 10.4 A reflection. The 0.48% elongation of the thin filament in the active state agrees with an elongation that is presumed by White and Thorson (1973) to estimate the lower limit of the thin-filament stiffness. It seems that in the catch state the activated and resting thin filament structures are intermixed. The activated parts of the thin filament are probably more elongated than the apparent value, 0.32%.

Actin Cytoskeleton↗

Evaluation of grip strength with a sustained maximal isometric contraction for 6 and 10 seconds.

The grip strength in which subjects sustained a maximal isometric contraction for 6 seconds (the 6-second test) was compared with that for 10 seconds (the 10-second test) to clarify a reliable sustained grip strength test procedure. Fifty healthy students (22+/-4 years, 25 men and 25 women) participated in this study. Strength was measured continuously by the Dexter (Cedaron Medical Inc.). The maximal grip strength, the peak time, and the momentary strength every second during a trial were evaluated. For both tests, the reliable maximal strength and a typical strength-time curve consisting of an early peak time and a decrease in the strength after the peak gradually over time were obtained. In the 6-second test, the momentary strength after 5 seconds that was 82+/-10 approximately 87+/-7% of the peak strength showed good reliability. This variable may be effective in assessing the ability to sustain maximal grip strength.

Adolescent↗

Mechanical behavior of skeletal muscle during intermittent voluntary isometric contractions in humans.

Changes in contractile speed and force-fusion properties were examined during repetitive isometric contractions with the knee extensors at three different target force levels. Seven healthy subjects were studied at target force levels of 30, 45, and 60% of their maximal voluntary contraction (MVC) force. Repeated 6-s contractions followed by 4-s rest were continued until exhaustion. Contractile speed was determined for contractions elicited by electrical stimulation at 1-50 Hz given during exercise and a subsequent 27-min recovery period. Contraction time remained unchanged during exercise and recovery, except for an initial rapid shift in the twitch properties. Half relaxation time (RT1/2) decreased gradually by 20-40% during exercise at 30 and 45% of MVC. In the recovery period, RT1/2 values were not fully restored to preexercise levels. During exercise at 60% MVC, the RT1/2 decreased for twitches and increased for the 50-Hz stimulation. In the recovery period after 60% MVC, RT1/2 values declined toward those seen after the 30 and 45% MVC exercise. The force oscillation amplitude in unfused tetani relative to the mean force increased during exercise at 30 and 45% MVC but remained unaltered during the 60% MVC exercise. This altered force-fusion was closely associated with the changes in RT1/2. The faster relaxation may at least partly explain the increased energy cost of contraction reported previously for the same type of exercise.

Adult↗

Demonstration of the significance of isometric contraction for the formation of stress fibres in chick embryo fibroblasts.

Observations of the cytoskeleton visualized with Coomassie brilliant blue R250 were carried out on chick embryo fibroblasts emigrating from cell aggregates onto clean or siliconized cover glasses. Conditions for persistent isometric contraction were created in cells attached strongly to glass and to the aggregate. These cells showed a presence of abundant stress fibres in their cytoplasm. Upon induction of lamellipodia retraction, the stress fibres transformed into typical retraction fibres. In cells spreading onto siliconized glass, the fibres were rare. Upon cell contraction their lamellipodia detached, folded and fibres, if originally present, disintegrated. These observations extend the conclusions of Wohlfarth-Bottermann to tissue culture cells, formulated on the basis of extensive study on the structure and behaviour of cytoplasmic fibrils in slime mold plasmodia, that bundles of parallely oriented actomyosin filaments arrange as a result of the persistent isometric contraction of cytoplasm. The present result point out the significance of cell surface adhesive properties for the organization of cell cytoskeleton.

Animals↗

Ultrastructure of clots during isometric contraction.

We explored the retraction or contraction of platelet-fibrin clots under isometric conditions. In the presence of micromolar calcium clots of normal platelet-rich plasma developed tension at an initial rate of 0.1 to 0.2 g/min per cm2 (initial cross-sectional area). Electron microscopy of clots fixed after attaining a force of 1.6 g/cm2 revealed platelets with elongated bodies and pseudopods in close apposition to fibrin strands which were oriented in cablelike fashion in the direction of tension. The development of tension could not be explained simply on the basis of platelet-platelet association and interaction alone. First, factor XIII-dependent cross-linking of fibrin fibers was critical to normal isometric contraction. Second, tension decreased linearly, rather than exponentially, when the platelet count in the platelet-fibrin clot was decreased, suggesting that platelets must be interacting with another component (i.e. fibrin). Thrombasthenic platelets, deficient in fibrinogen receptors, failed to develop tension or to align fibrin strands or pseudopods in the clot. Platelet-fibrin clots treated with vincristine to disassemble microtubules or cytochalasin B to disrupt microfilaments failed to develop tension and relaxed if these agents were added after tension had developed. Relaxation under these conditions, however, was not associated with loss of orientation of fibrin strands. Our findings suggest that platelet-fibrin interaction in clots under isometric conditions leads to orientation of fibrin strands and platelets in the direction of force generation. Tension develops as platelets simultaneously attach to and spread along fibrin strands, and contract. The contraction draws some fibrin into platelet-fibrin clumps and aligns other strands in the long axis of tension. The achievement and maintenance of maximum tension appears to depend on the development of platelet-fibrin attachments and extension of platelet bodies and long pseudopods containing bundles of microfilaments and microtubules along the oriented fibrin fibers.

Blood Platelets↗