Relationships between force curves and muscle oxygenation kinetics during repeated handgrip

The purpose of this study was to clarify the kinetics of muscle oxygenation by near infrared spectroscopy (NIRS) in the phase of the decreasing force, especially the pre- and post-phases of the inflection point, during repeated rhythmic grip (RRG) of 30 grips/min(-1) for 6 minutes. The inflection point was the time at which the decreasing speed of the grip force changed markedly. It was calculated statistically from two regression lines fitted to each decreasing phase by applying a two-phase regression model. Ten healthy males performed the RRG for 6 minutes. Total Hb and Oxy-Hb decreased rapidly about 10 sec (7.0+/-5.9 sec, 9.8+/-5.4 sec, respectively) corresponding to the value decreasing by 90% MVC after the onset of gripping. Deoxy-Hb was maintained at a high value for 76.2+/-27.9 sec, corresponding to the value decreasing by 70-80% MVC. These phases are considered to be the states where oxygen was not satisfactorily supplied to the active muscles because of the obstruction of blood flow caused by an increase in the intra-muscular pressure. Deoxy-Hb decreased for 120+/-21.3 sec after reaching the highest value, and then reached an almost steady state at a higher level than the rest. After this phase, muscle oxygenation kinetics enters the state where oxygen is satisfactorily supplied to active muscles. We considered that the relationship between oxygen supply and demand differs during the initial and the latter phases in RRG. The changing phase in the decreasing speed of the grip force, namely the inflection point of the decreasing force, significantly correlated with the changing phase of the Oxy-Hb and Deoxy-Hb kinetics. The inflection point of the decreasing force seems to correspond to the phase where oxygen supply cannot meet oxygen demand and the increase of Deoxy-Hb. We infer that the pre- and post-phases of the inflection point depend on different physiological factors.     

The influence of different target values and measurement times on the decreasing force curve during sustained static gripping work

The purposes of this study were to clarify the decreasing properties of, and to examine useful measurement times for evaluating muscle endurance in a comparison among various parameters using measurement times of 1, 3 and 6 mins and target values of 50, 75 and 100% MVC. Fifteen males and 15 females participated in this study. All subjects carried out sustained isometric gripping under nine conditions of measurement times and target forces, (1, 3 and 6 mins vs. 50, 75 and 100% MVC) with an interval of one or two days. The property of decreasing force in the initial phase (marked decreasing phase) differed among the target values, and the decreasing speed of the gripping force was highest for 100% MVC. However, the decreasing property after about 60 sec, in which the force decreased to about 30% MVC from the onset of grip, was similar among all target values, and then the gripping force reached an almost steady state phase at about 150-180 sec. In other words, the difference of the decreasing property during the initial phase with different target values was considered not to influence the property in the later phase, in which the force decreases to about 30% MVC. When muscle endurance is evaluated from the phase until reaching the steady state, it may be possible to evaluate the same property of the decreasing phase at 6 min as the measurement at 3 min. The measurement for 1 min at 50% MVC was not valid as an evaluation time because the grip force did not decrease enough. The integrated area in the initial phase was considered to depend on the magnitude of the target value, and the integrated area for 30 sec or 60 sec at 75% MVC was larger than that at 100% MVC. It was inferred that higher pain at 100% MVC resulted in a greater decrease in the speed of the force.     

The effect of measurement time when evaluating static muscle endurance during sustained static maximal gripping

The purpose of this study was to examine the useful measurement time when evaluating static muscle endurance by comparing various parameters during sustained static gripping for 1, 3 and 6 min. Fifteen males (mean +/- SD age 20.8 +/- 1.3 yr, height 172.9 +/- 4.6 cm, body mass 67.7 +/- 5.7 kg) and fifteen females [mean +/- SD age 20.2 +/- 0.9 yr, height 158.5 +/- 3.2 cm, body mass 55.9 +/- 4.6 kg] volunteered to participate in this study. The subjects performed the sustained static maximal grip test with a sagittal and horizontal arm position for 1, 3 and 6 min on different days. Eleven force-time parameters were selected to evaluate static muscle endurance. The trial-to-trial reliability of each measurement time of sustained static maximal gripping was very high (rxy = 0.887-0.981 (1 min), 0.912-0.993 (3 min), 0.901-0.965 (6 min)). The errors of exertion values between trials were very small (below 10%). A significant correlation was found in the following parameters: the final strength and the exponential function between 1 min and 3 min, all parameters except for the time required to reach 80% of maximal grip, the regression coefficient at post-inflection between 3 min and 6 min, and the decreasing rate between all measurement times (1 min, 3 min, and 6 min). Significant differences between the measurement times were found in all parameters except for the time to 60, 70, and 80% force decreases, and the regression coefficient of pre-inflection. There was a tendency that the longer the measurement time, the larger the decreasing force. It is suggested that for the 6 min measurement, the subjects unconsciously restrained the maximal gripping force, influenced by a psychological factor as the pain became greater. The 1 min measurement may evaluate only the remarkable decreasing phase of the decreasing force, and not evaluate the phase of an almost steady state.     

Relationships between force-time parameters and muscle oxygenation kinetics during maximal sustained isometric grip and maximal repeated rhythmic grip with different contraction frequencies

The purposes of this study were to examine the relationships between various force-time parameters and muscle oxygenation kinetics during maximal sustained isometric grip (SIG) and maximal repeated rhythmic grips (RRG) with different grip intervals (interval times: 5, 4, 3, and 2 s). Subjects were 10 healthy young males, aged 20-26 years (height 173.9+/-7.3 cm, body mass 71.5+/-11.2 kg). After measuring maximal grip force, each subject performed the SIG and RRG tests with a target frequency of 12, 15, 20, and 30 grips.min(-1) (interval times: 5, 4, 3, and 2 s, respectively) for 6 min. The decreasing time until 80% MVC showed significant and high correlations with final force values in RRGs with over 3 s intervals (r=0.866-0.941), but not in the SIG and RRG with a 2 s interval. The time at the lowest Oxy-Hb/Mb value showed a significant and high correlation with the time at the highest Deoxy-Hb/Mb value only in the SIG and RRG with a 2 s interval (r=0.825-0.916). Oxy-Hb/Mb decreases markedly and deoxy-Hb/Mb increases after the onset of SIG due to the obstruction of blood flow caused by the increase in intramuscular pressure. A similar physiological response to that of SIG occurs also in RRG with a 2 s interval, but RRGs with intervals over 3 s achieve more resumption of blood flow in the muscular relaxation phase. Hence, in spite of the same RRGs, it was determined that RRGs with intervals over 3 s differ significantly in a changing pattern of grip force and muscle oxygen kinetics from RRGs with a 2 s interval.     

A comparison of static and dynamic optimization muscle force predictions during wheelchair propulsion

The primary purpose of this study was to compare static and dynamic optimization muscle force and work predictions during the push phase of wheelchair propulsion. A secondary purpose was to compare the differences in predicted shoulder and elbow kinetics and kinematics and handrim forces. The forward dynamics simulation minimized differences between simulated and experimental data (obtained from 10 manual wheelchair users) and muscle co-contraction. For direct comparison between models, the shoulder and elbow muscle moment arms and net joint moments from the dynamic optimization were used as inputs into the static optimization routine. RMS errors between model predictions were calculated to quantify model agreement. There was a wide range of individual muscle force agreement that spanned from poor (26.4% F_max error in the middle deltoid) to good (6.4% F_max error in the anterior deltoid) in the prime movers of the shoulder. The predicted muscle forces from the static optimization were sufficient to create the appropriate motion and joint moments at the shoulder for the push phase of wheelchair propulsion, but showed deviations in the elbow moment, pronation–supination motion and hand rim forces. These results suggest the static approach does not produce results similar enough to be a replacement for forward dynamics simulations, and care should be taken in choosing the appropriate method for a specific task and set of constraints. Dynamic optimization modeling approaches may be required for motions that are greatly influenced by muscle activation dynamics or that require significant co-contraction.     

Effect of work and recovery duration on skeletal muscle oxygenation and fuel use during sustained intermittent exercise

The purpose of this study was to compare rates of substrate oxidation in two protocols of intermittent exercise, with identical treadmill speed and total work duration, to reduce the effect of differences in factors such as muscle fibre type activation, hormonal responses, muscle glucose uptake and non-esterified fatty acid (NEFA) availability on the comparison of substrate utilisation. Subjects (n = 7) completed 40 min of intermittent intense running requiring a work:recovery ratio of either 6 s:9 s (short-interval exercise, SE) or 24 s:36 s (long-interval exercise, LE), on separate days. Another experiment compared O(2) availability in the vastus lateralis muscle across SE (10 min) and LE (10 min) exercise using near-infrared spectroscopy (RunMan, NIM. Philadelphia, USA). Overall (i.e. work and recovery) O(2) consumption (VO(2)) and energy expenditure were lower during LE (P < 0.01, P < 0.05, respectively). Overall exercise intensity, represented as a proportion of peak aerobic power (VO2(peak)), was [mean (SEM)] 64.9 (2.7)% VO2(peak) (LE) and 71.4 (2.4)% VO2(peak) (SE). Fat oxidation was three times lower (P < 0.01) and carbohydrate oxidation 1.3 times higher (P < 0. 01) during LE, despite the lower overall exercise intensity. Plasma lactate was constant and was higher throughout exercise in LE [mean (SEM) 5.33 (0.53) mM, LE; 3.28 (0.31) mM, SE; P < 0.001)]. Plasma pyruvate was higher and glycerol was lower in LE [215 (17) microM, 151 (13) microM, P < 0.05, pyruvate; 197 (19) microM, 246 (19) microM, P < 0.05, glycerol]. There was no difference between protocols for plasma NEFA concentration (n = 4) or plasma noradrenaline and adrenaline. Muscle oxygenation declined in both protocols (P < 0.001), but the nadir during LE was lower [52.04 (0. 60)%] compared to SE [61.85 (0.51)%; P < 0.001]. The decline in muscle oxygenation during work was correlated with mean lactate concentration (r = 0.68; P < 0.05; n = 12). Lower levels of fat oxidation occurred concurrent with accelerated carbohydrate metabolism, increases in lactate and pyruvate and reduced muscle O(2) availability. These changes were associated with proportionately longer work and recovery periods, despite identical treadmill speed and total work duration. The proposal that a metabolic regulatory factor within the muscle fibre retards fat oxidation under these conditions is supported by the current findings.     

Skeletal muscle oxygenation during incremental exercise

The purpose of this study was to investigate the relationship between muscle oxygenation level at exhaustion and maximal oxygen uptake (VO2max) in an incremental cycling exercise. Nine male subjects took part in an incremental exhaustive cycling exercise, and then cuff occlusion was performed. Changes in oxy-(deltaHbO2) and deoxy-(deltaHb) hemoglobin concentrations in the vastus lateralis muscle were measured with a near infrared spectroscopy (NIRS). Muscle oxygenation during incremental exercise was expressed as a percentage (%Moxy) of the maximal range observed during an arterial occlusion as the lower reference point. A systematic decrease was observed in %Moxy with increasing intensity. A significant relationship was observed between %Moxy at exhaustion and VO2max (p < 0.01). We concluded that the one of the limiting factor of VO2max is the muscle oxygen diffusion capacity, and %Moxy during exercise could be one of the indexes of muscle oxygen diffusion capacity.     

The effect of decreasing erythrocyte membrane permeability for oxygen during oxygenation

It was shown that erythrocyte membranes permeability for oxygen decreases at least a few tens of times during oxygenation.     

Relationship between oxygenation in inactive biceps brachii muscle and hyperventilation during leg cycling

Inactive forearm muscle oxygenation has been reported to begin decreasing from the respiratory compensation point (RCP) during ramp leg cycling. From the RCP, hyperventilation occurs with a decrease in arterial CO2 pressure (PaCO2). The aim of this study was to determine which of these two factors, hyperventilation or decrease in PaCO2, is related to a decrease in inactive biceps brachii muscle oxygenation during leg cycling. Each subject (n = 7) performed a 6-min two-step leg cycling. The exercise intensity in the first step (3 min) was halfway between the ventilatory threshold and RCP (170+/-21 watts), while that in the second step (3 min) was halfway between the RCP and peak oxygen uptake (240+/-28 watts). The amount of hyperventilation and PaCO2 were calculated from gas parameters. The average cross correlation function in seven subjects between inactive muscle oxygenation and amount of hyperventilation showed a negative peak at the time shift of zero (r = -0.72, p<0.001), while that between inactive muscle oxygenation and calculated PaCO2 showed no peak near the time shift of zero. Thus, we concluded that decrease in oxygenation in inactive arm muscle is closely coupled with increase in the amount of hyperventilation.     

An EMG fractal indicator having different sensitivities to changes in force and muscle fatigue during voluntary static muscle contractions.

During a sustained contraction, electromyographic signals (EMGs) undergo a spectral compression. This fatigue behaviour induces a shift of the mean and the median frequencies to lower frequencies. On the other hand, several studies conclude that the mean/median frequency can increase, decrease or remain constant with an increasing force level. Such inconsistency is embarrassing since the fatigue state may be influenced by the force level. In this paper, we propose a frequency indicator which is sensitive to the force level independently of the fatigue state evaluated at 70% of the maximal voluntary contraction. Ten healthy volunteers participated in the study and both surface EMGs (from the short head of the biceps brachii) and force signals were measured. This study compared force and fatigue effects on the EMGs during short (3-s) isometric contractions at different strength intensities and during a sustained isometric contraction until exhaustion. The EMGs partly show 1/falpha spectral behaviours since their power spectral densities may experimentally fit with two linear segments in a log-log representation. The measured "right" slope produces variations of force as 20 times the variations of fatigue. 1/falpha Behaviour may be related to stochastic fractals. This fractal indicator is a new frequency indicator that is thus complementary to other known classical frequency indicators when studying force during unknown fatigue states.     

Targeted gripping reduces shoulder muscle activity and variability

The purpose of this study was to determine if the effect of visually targeted gripping on shoulder muscle activity was maintained with repeated exposures. Eleven healthy males had eight shoulder muscles monitored via surface electromyography while maintaining shoulder elevation at 90° in the scapular plane with and without a 30% grip force. Three non-gripping trials were followed by 15 gripping trials and another 3 non-gripping control trials. Gripping significantly decreased the activity of the anterior deltoid, trapezius, and latissimus dorsi over the exposure of 15 trials. Gripping also reduced variability in all muscles' activity. The changes in shoulder muscle activity are likely in response to forces being transferred through multi-articular muscles spanning from the forearm to the shoulder. Targeted gripping during shoulder elevation resulted in small but significant decreases in muscle activity and reduced variability which supports previous evidence for increased risk of upper extremity disorders in occupational settings.     

Changes in muscle oxygenation during weight-lifting exercise

The quantitative analysis of haemoglobin oxygenation of contracting human muscle during weight-lifting exercise was studied noninvasively and directly using near-infrared spectroscopy. This method was developed as a three-wavelength method which confirmed the volume changes in oxygenated haemoglobin (oxy-Hb), deoxygenated haemoglobin (deoxy-Hb) and blood volume (total-Hb; Oxy-Hb + deoxy-Hb). Nine healthy adult men with various levels of training experience took part in the study. Ten repetition maximum (10 RM) one-arm curl exercise was performed by all the subjects. Results showed that at the beginning of the 10-RM exercise, rapid increases of deoxy-Hb and decreases of oxy-Hb were observed. In addition, total-Hb gradually increased during exercise. These results corresponded to the condition of arm blood flow experimentally restricted using a tourniquet in contact with the shoulder joint, and they showed the restriction of venous blood flow and an anoxic state occurring in the dynamically contracted muscle. In three sets of lifting exercise with short rest periods, these tendencies were accelerated in each set, while total-Hb volume did not return to the resting state after the third set for more than 90 s. These results would suggest that a training regimen emphasizing a moderately high load and a high number of repetitions, and a serial set with short rest periods such as usually performed by bodybuilders, caused a relatively long-term anoxic state in the muscle.     

Biceps brachii myoelectric and oxygenation changes during static and sinusoidal isometric exercises

Surface myoelectric signal changes occurring during sustained isometric contractions have been extensively studied with quantitative surface electromyography (sEMG) and are described by means of some sEMG global variables in time and frequency domain (such as the median power spectral frequency). Recently, the possibility of studying local muscle O₂ saturation during exercise using non-invasive methods has been enhanced thanks to the use of near-infrared spectroscopy (NIRS). The purpose of this work was to combine NIRS and sEMG techniques to analyze the relationship between modifications of sEMG parameters and the underlying metabolic status of the exercising biceps brachii muscle. This relationship was tested under different isometric contraction modalities, namely static (ST) at 20, 40, 60 and 80%MVC and sinusoidal (SIN) at 40 ± 20 and 60 ± 20%MVC. Results clearly indicated the presence of an initial fast phase of muscle O₂ desaturation followed by a slow phase, regardless of the contraction modality. Moreover, the initial rate of muscle O₂ desaturation was related to the level of force output (R = 0.92), but it was independent on the contraction modality (p < 0.05). Similarly, changes in sEMG parameters were related to force level (Conduction Velocity-CV vs. Force: R = 0.87; sEMG Median Frequency-MDF vs. Force: R = 0.86). The high correlation found between CV-MDF and Tissue Oxygenation Index (TOI) slope (R = 0.73 and 0.72, respectively) suggests a strong relationship between NIRS and sEMG data. This study indicates that muscle O₂ demand during isometric contractions from low to high force levels is influenced by the type of active motor units and not from the type of isometric exercise modality.     

The kinetics relating calcium and force in skeletal muscle.

The kinetics relating Ca2+ transients and muscle force were examined using data obtained with the photoprotein aequorin in skeletal muscles of the rat, barnacle, and frog. These data were fitted by various models using nonlinear methods for minimizing the least mean square errors. Models in which Ca2+ binding to troponin was rate limiting for force production did not produce good agreement with the observed data, except for a small twitch of the barnacle muscle. Models in which cross-bridge kinetics were rate limiting also did not produce good agreement with the observed data, unless the detachment rate constant was allowed to increase sharply on the falling phase of tension production. Increasing the number of cross-bridge states did not dramatically improve the agreement between predicted and observed force. We conclude that the dynamic relationship between Ca2+ transients and force production in intact muscle fibers under physiological conditions can be approximated by a model in which (a) two Ca2+ ions bind rapidly to each troponin molecule, (b) force production is limited by the rate of formation of tightly bound cross-bridges, and (c) the rate of cross-bridge detachment increases rapidly once tension begins to decline and free Ca2+ levels have fallen to low values after the last stimulus. Such a model can account not only for the pattern of force production during a twitch and tetanus, but also the complex, nonlinear pattern of summation which is observed during an unfused tetanus at intermediate rates of stimulation.     

Alternate muscle activity observed between knee extensor synergists during low-level sustained contractions.

To determine quantitatively the features of alternate muscle activity between knee extensor synergists during low-level prolonged contraction, a surface electromyogram (EMG) was recorded from the rectus femoris (RF), vastus lateralis (VL), and vastus medialis (VM) in 11 subjects during isometric knee extension exercise at 2.5% of maximal voluntary contraction (MVC) for 60 min (experiment 1). Furthermore, to examine the relation between alternate muscle activity and contraction levels, six of the subjects also performed sustained knee extension at 5.0, 7.5, and 10.0% of MVC (experiment 2). Alternate muscle activity among the three muscles was assessed by quantitative analysis on the basis of the rate of integrated EMG sequences. In experiment 1, the number of alternations was significantly higher between RF and either VL or VM than between VL and VM. Moreover, the frequency of alternate muscle activity increased with time. In experiment 2, alternating muscle activity was found during contractions at 2.5 and 5.0% of MVC, although not at 7.5 and 10.0% of MVC, and the number of alternations was higher at 2.5 than at 5.0% of MVC. Thus the findings of the present study demonstrated that alternate muscle activity in the quadriceps muscle 1) appears only between biarticular RF muscle and monoarticular vasti muscles (VL and VM), and its frequency of alternations progressively increases with time, and 2) emerges under sustained contraction with force production levels < or =5.0% of MVC.     

A comparison of skeletal muscle oxygenation and fuel use in sustained continuous and intermittent exercise

In this study we compared substrate oxidation and muscle oxygen availability during sustained intermittent intense and continuous submaximal exercise with similar overall (i.e. work and recovery) oxygen consumption (VO2). Physically active subjects (n = 7) completed 90 min of an intermittent intense (12 s work:18 s recovery) and a continuous submaximal treadmill running protocol on separate days. In another experiment (n = 5) we compared oxygen availability in the vastus lateralis muscle between these two exercise protocols using near-infrared spectroscopy. Initially, overall VO(2) (i.e. work and recovery) was matched, and from 37.5 min to 67.5 min of exercise was similar, although slightly higher during continuous exercise (8%; P < 0.05). Energy expenditure was constant (22.5-90 min of exercise) and was not different in intermittent intense [0.81 (0.01) kJ x min(-1). kg(-1)] and continuous submaximal [0.85 (0.01) kJ x min(-1) x kg(-1)] exercise. Overall exercise intensity, represented as a proportion of peak aerobic power (VO2(peak)), was 68.1 (2.5)% VO2(peak) and 71.8 (1.8)% VO2(peak) for intermittent and continuous exercise protocols, respectively. Fat oxidation was almost 3 times lower (P < 0.05) and carbohydrate oxidation was approximately 1.2 times higher (P < 0.05) during intermittent compared to continuous exercise, despite the same overall energy expenditure. Capillary plasma lactate was constant from 15 to 90 min of exercise, and pyruvate was constant from 15 to 75 min, although both were higher (P < 0.0001, lactate; P < 0.001, pyruvate) during intermittent [5.05 (0.28) mM, 200 (7) microM, respectively] compared to continuous exercise [2.41 (0.10) mM, 114 (4) microM, respectively]. There was no difference between protocols for either plasma glycerol or non-esterified fatty acids. The decrease in muscle oxygenation during work periods of intermittent exercise resulted in a lower nadir oxygenation [54.62 (0.41)%] compared to continuous exercise [58.82 (0.21)%, P < 0.001]. The decline in oxygenation was correlated with treadmill speed (r = 0.72; P < 0.05). These results show a difference in substrate utilisation and muscle oxygen availability during sustained intermittent intense and continuous submaximal exercise, despite a similar overall VO(2) and identical energy expenditure.     

Relationships between nonhyperbolic kinetics and dimeric structure in ribonucleases

Dimers of bovine pancreatic RNase A give nonhyperbolic saturation curves for the substrate of the second, rate-limiting step of the reaction. Under the same conditions, the monomeric native enzyme shows Michaelis-Menten kinetics. Naturally dimeric bovine seminal RNase, which has been found to give nonhyperbolic saturation curves, loses this property upon monomerization. It is proposed that when RNase monomers are arranged in a quaternary structure, they assume a conformation which enables them to be modulated in their catalytic activities. A correlation is suggested between this effect and the quaternary structure proposed for both of these dimeric ribonucleases, in which composite active sites are generated by the mutual exchange of the NH2-terminal ends of the two monomers.     

Duration of static stretching influences muscle force production in hamstring muscles

The purpose of the present study was to investigate whether duration of static stretching could affect the maximal voluntary contraction (MVC). Volunteer male subjects (n = 10) underwent 2 different durations of static stretching of their hamstring muscles in the dominant leg: 30 and 60 seconds. No static stretching condition was used as a control condition. Before and after each stretching trial, hamstring flexibility was measured by a sit and reach test. MVC was then measured using the maximal effort of knee flexion. The hamstring flexibility was significantly increased by 30 and 60 seconds of static stretching (control: 0.5 +/- 1.1 cm; 30 seconds: 2.1 +/- 1.8 cm; 60 seconds: 3.0 +/- 1.6 cm); however, there was no significant difference between 30 and 60 seconds of static stretching conditions. The MVC was significantly lowered with 60 seconds of static stretching compared to the control and 30 seconds of the stretching conditions (control: 287.6 +/- 24.0 N; 30 seconds: 281.8 +/- 24.2 N; 60 seconds: 262.4 +/- 36.2 N). However, there was no significant difference between control and 30 seconds of static stretching conditions. Therefore, it was concluded that the short duration (30 seconds) of static stretching did not have a negative effect on the muscle force production.     

Characterization of cross-bridge elasticity and kinetics of cross-bridge cycling during force development in single smooth muscle cells

Force development in smooth muscle, as in skeletal muscle, is believed to reflect recruitment of force-generating myosin cross-bridges. However, little is known about the events underlying cross-bridge recruitment as the muscle cell approaches peak isometric force and then enters a period of tension maintenance. In the present studies on single smooth muscle cells isolated from the toad (Bufo marinus) stomach muscularis, active muscle stiffness, calculated from the force response to small sinusoidal length changes (0.5% cell length, 250 Hz), was utilized to estimate the relative number of attached cross-bridges. By comparing stiffness during initial force development to stiffness during force redevelopment immediately after a quick release imposed at peak force, we propose that the instantaneous active stiffness of the cell reflects both a linearly elastic cross-bridge element having 1.5 times the compliance of the cross-bridge in frog skeletal muscle and a series elastic component having an exponential length-force relationship. At the onset of force development, the ratio of stiffness to force was 2.5 times greater than at peak isometric force. These data suggest that, upon activation, cross-bridges attach in at least two states (i.e., low-force-producing and high-force-producing) and redistribute to a steady state distribution at peak isometric force. The possibility that the cross-bridge cycling rate was modulated with time was also investigated by analyzing the time course of tension recovery to small, rapid step length changes (0.5% cell length in 2.5 ms) imposed during initial force development, at peak force, and after 15 s of tension maintenance. The rate of tension recovery slowed continuously throughout force development following activation and slowed further as force was maintained. Our results suggest that the kinetics of force production in smooth muscle may involve a redistribution of cross-bridge populations between two attached states and that the average cycling rate of these cross-bridges becomes slower with time during contraction.     

Impact of detubulation on force and kinetics of cardiac muscle contraction

Action potential–driven Ca²⁺ currents from the transverse tubules (t-tubules) trigger synchronous Ca²⁺ release from the sarcoplasmic reticulum of cardiomyocytes. Loss of t-tubules has been reported in cardiac diseases, including heart failure, but the effect of uncoupling t-tubules from the sarcolemma on cardiac muscle mechanics remains largely unknown. We dissected intact rat right ventricular trabeculae and compared force, sarcomere length, and intracellular Ca²⁺ in control trabeculae with trabeculae in which the t-tubules were uncoupled from the plasma membrane by formamide-induced osmotic shock (detubulation). We verified disconnection of a consistent fraction of t-tubules from the sarcolemma by two-photon fluorescence imaging of FM4-64–labeled membranes and by the absence of tubular action potential, which was recorded by random access multiphoton microscopy in combination with a voltage-sensitive dye (Di-4-AN(F)EPPTEA). Detubulation reduced the amplitude and prolonged the duration of Ca²⁺ transients, leading to slower kinetics of force generation and relaxation and reduced twitch tension (1 Hz, 30°C, 1.5 mM [Ca²⁺]_o). No mechanical changes were observed in rat left atrial trabeculae after formamide shock, consistent with the lack of t-tubules in rodent atrial myocytes. Detubulation diminished the rate-dependent increase of Ca²⁺-transient amplitude and twitch force. However, maximal twitch tension at high [Ca²⁺]_o or in post-rest potentiated beats was unaffected, although contraction kinetics were slower. The ryanodine receptor (RyR)2 Ca-sensitizing agent caffeine (200 µM), which increases the velocity of transverse Ca²⁺ release propagation in detubulated cardiomyocytes, rescued the depressed contractile force and the slower twitch kinetics of detubulated trabeculae, with negligible effects in controls. We conclude that partial loss of t-tubules leads to myocardial contractile abnormalities that can be rescued by enhancing and accelerating the propagation of Ca²⁺-induced Ca²⁺ release to orphan RyR2 clusters.