Residual force enhancement after stretch in striated muscle. A consequence of increased myofilament overlap?

When skeletal muscle is stretched above optimal sarcomere length during tetanic activity there is an increase in force that stays above the isometric force level throughout the activity period. This long-lasting increase in contractile force, generally referred to as 'residual force enhancement after stretch' (FE(resid)), has been studied in great detail in various muscle preparations over more than half a century. Substantial evidence has been presented to show that non-uniform sarcomere behaviour plays a major part in the development of FE(resid). However, in a great number of recent studies the role of sarcomere non-uniformity has been challenged and alternative mechanisms have instead been proposed to explain the increase in force such as enhancement of cross-bridge function and/or strengthening of parallel elastic elements along the muscle fibres. This article presents a short review of the salient features of FE(resid) and provides evidence that non-uniform sarcomere behaviour is indeed likely to play a major role in the development of FE(resid). Electron microscopical studies of fibres rapidly fixed after active stretch demonstrate that, dispersed in the preparation, there are assymetrical length changes within the two halves of myofibrillar sarcomeres resulting in greater filament overlap in one half of the sarcomere than in the opposite sarcomere half. Sarcomere halves with increased filament overlap will consequently be in a situation where they are able to produce a greater force than that recorded in the isometric control. Weaker regions in series will be able to keep the enhanced force by recruitment of elastic elements.

Contractile performance of striated muscle.

The single muscle fiber preparation provides an excellent tool for studying the mechanical behaviour of the contractile system at sarcomere level. The present article gives an overview of studies based on intact single fibers from frog and mouse skeletal muscle. The following aspects of muscle function are treated: (1) The length-tension relationship. (2) The biphasic force-velocity relationship. (3) The maximum speed of shortening, its independence of sarcomere length and degree of activation. (4) Force enhancement during stretch, its relation to sarcomere length and myofilament lattice width. (5) Residual force enhancement after stretch. (6) Force reduction after loaded shortening. (7) Deactivation by active shortening. (8) Differences in kinetic properties along individual muscle fibers.

Release of calcium into the myofibrillar space in response to active shortening of striated muscle.

AIM: The study was undertaken to explore whether shortening of striated muscle during activity is associated with release of bound Ca2+ into the myofibrillar space as has previously been proposed in order to explain the depressant effect of active shortening. METHODS: The experiments were carried out on single muscle fibres isolated from the anterior tibialis muscle of Rana temporaria. The fibres were loaded with the calcium sensitive indicator Fluo-3. The fibres, stimulated to produce a partially fused isometric tetanus, were subjected to a shortening ramp or, alternatively, to a stretch ramp during activity while force, fibre length, sarcomere length and the Fluo-3 signal were recorded. RESULTS: A shortening ramp performed during a partially fused tetanus caused an increase in the myofibrillar free calcium concentration and produced, simultaneously, a decrease in active force. The isometric force recovered gradually after the shortening ramp, while the intracellular Ca2+ concentration stayed above the control level during the remainder of the stimulation period. A stretch ramp applied during a partially fused tetanus caused a considerably smaller change in the myofibrillar Ca2+ concentration. CONCLUSION: The results provide evidence that the myosin cross-bridges interact with the calcium binding sites on the thin filaments during active shortening, causing sustained release of calcium and reduced contractile strength.

The force-velocity relationship at negative loads (assisted shortening) studied in isolated, intact muscle fibres of the frog.

AIM: The study was undertaken to explore the force-velocity relationship under conditions where the myofilament system is subjected to an external force that serves as a negative load and assists the shortening movement. METHODS: The experiments were carried out on single muscle fibres isolated from the anterior tibialis muscle of Rana temporaria. The fibres, being operated under load-clamp control, were released to shorten during tetanic stimulation at sarcomere lengths where the fibres carried different degrees of passive tension. The shortening thus occurred while the sarcomeres were subjected to a force that may be characterized as a 'negative load', that is, a force assisting the shortening movement. RESULTS: The force-velocity relationship below zero load was found to be a smooth continuation of the force-velocity curve recorded at positive loads with the shortening velocity increasing steeply at loads below zero. A negative load amounting to merely 10% of the isometric force, thus raised the shortening velocity to a level two to three times higher than V0 , the velocity recorded at zero load. CONCLUSIONS: The results provide evidence that, even in the presence of a longitudinal compressive force, the speed of shortening of the muscle fibre is determined by the cycling rate of the interacting cross-bridges. The force-velocity relationship at negative loads may play a relevant part during fast movements of striated muscle as pointed out in the discussion.

Non-linear myofilament elasticity in frog intact muscle fibres.

The aim of the present investigation was to elucidate the elastic properties of the myofilaments during tetanic activity in striated muscle. The study was carried out on intact single muscle fibres from the anterior tibialis muscle of Rana temporaria (2.0-2.5 degrees C). The instantaneous stiffness was measured as the change in force that occurred in response to a high-frequency (2-4 kHz) length oscillation while the fibre was released to shorten against a pre-set constant load that ranged between 40 and 70% of maximum tetanic force in different experiments. Measurements of fibre stiffness were carried out, at a given load, both at 2.20 microm sarcomere length (S(2.20)), i.e. at full overlap between the thick and thin filaments, and at 2.60 microm sarcomere length (S(2.60)). The fact that the load on the fibre was constant during the stiffness measurements at the two sarcomere lengths implies that the stiffness of elastic elements, acting in series with the myofilaments, was constant at the two sarcomere lengths. The fibre stiffness was consistently lower at the extended sarcomere length, the S(2.60)/S(2.20) ratio ranging from 0.83 to 0.97 at the different loads investigated. Based on the S(2.60)/S(2.20) ratio, the compliance of the free portions of the thick and thin filaments could be calculated. The myofilament stiffness was found to increase progressively as the load was raised from 40 to 70% of maximum tetanic force. At 2.20 microm sarcomere length and at 40% of maximum load on the fibre, the calculated myofilament stiffness was approximately 2.5 times the maximum cross-bridge stiffness.

Determinants of force rise time during isometric contraction of frog muscle fibres.

Force-velocity (F-V) relationships were determined for single frog muscle fibres during the rise of tetanic contraction. F-V curves obtained using isotonic shortening early in a tetanic contraction were different from those obtained at equivalent times with isovelocity shortening, apparently because changing activation early in the contraction leads, in isovelocity experiments, to changing force and changing series elastic extension. F-V curves obtained with isotonic and with isovelocity shortening are similar if the shortening velocity in the isovelocity trials is corrected for series elastic extension. There is a progressive shift in the scaling of force-velocity curves along the force axis during the course of the tetanic rise, reflecting increasing fibre activation. The time taken for F-V curves to reach the steady-state position was quite variable, ranging from about 50 ms after the onset of contraction (1-3 degrees C) to well over 100 ms in different fibres. The muscle force at a fixed, moderately high shortening velocity relative to the force at this velocity during the tetanic plateau was taken as a measure of muscle activation. The reference velocity used was 60% of the maximum shortening velocity (V(max)) at the tetanic plateau. The estimated value of the fractional activation at 40 ms after the onset of contraction was used as a measure of the rate of activation. The rate of rise of isometric tension in different fibres was correlated with the rate of fibre activation and with V(max) during the plateau of the tetanus. Together differences in rate of activation and in V(max) accounted for 60-80% of the fibre-to-fibre variability in the rate of rise of isometric tension, depending on the measure of the force rise time used. There was not a significant correlation between the rate of fibre activation and V(max). The steady-state F-V characteristics and the rate at which these characteristics are achieved early in contraction are seemingly independent. A simulation study based on F-V properties and series compliance in frog muscle fibres indicates that if muscle activation were instantaneous, the time taken for force to rise to 50% of the plateau value would be about 60% shorter than that actually measured from living fibres. Thus about 60% of the force rise time is a consequence of the time course of activation processes and about 40% represents time taken to stretch series compliance by activated contractile material.

Contractile properties of mouse single muscle fibers, a comparison with amphibian muscle fibers.

Single fibers, 25-40 microm wide and 0.5-0.7 mm long, were isolated from the flexor digitorum brevis muscle of the mouse. Force and movement were recorded (21-27 degrees C) from the fiber as a whole and, in certain experiments, from a short marked segment that was held at constant length by feedback control. The maximum tetanic force, 368+/-57 kN/m2 (N = 10), was not significantly different from that recorded in frog muscle fibers at equal temperature. However, the rising phase of the tetanus was considerably slower in the mammalian fibers, 202+/-20 ms (N = 17) being required to reach 90% of maximum tetanic force as compared with 59+/-4 ms (N = 20) in the frog muscle fibers. Similar to the situation in frog muscle fibers, the force-velocity relation exhibited two distinct curvatures located on either side of a breakpoint near 80% of the isometric force. Maximum speed of shortening was 4.0+/-0.3 fiber lengths s(-1) (N = 6). The relationship between tetanic force and sarcomere length was studied between 1.5 and 4.0 microm sarcomere spacings, based on length-clamp recordings that were free of 'tension creep'. There was a flat maximum (plateau) of the length-tension relation between approximately 2.0 and 2.4 microm sarcomere lengths. The descending limb of the length-tension relation (linear regression) intersected the length axis (zero force) at 3.88 microm and reached maximum force at 2.40 microm sarcomere length. The slope of the descending limb is compatible with a thick filament length of 1.63 microm and an average thin filament length of 1.10 microm. These values accord well with recent electron microscope measurements of myofilament length in mammalian muscle.

Effects of intracellular acidification and varied temperature on force, stiffness, and speed of shortening in frog muscle fibers.

This study aimed to establish whether the temperature-dependent effect of acidification on maximum force observed in mammalian muscles also applies to frog muscle. Measurements of force, stiffness, and unloaded velocity of shortening in intact single muscle fibers from the anterior tibialis muscle of Rana temporaria were performed between 0 and 22 degrees C during fused tetani in H(2)CO(3)-CO(2)-buffered Ringer solution with pH adjusted to 7.0 and 6.3, respectively. The force-to-stiffness ratio increased as a rectilinear function of temperature between 0 and 20 degrees C at pH 7.0. Lowering the pH to 6.3 reduced the tetanic force by 13.5 +/- 1.2 and 11.5 +/- 1.4% at 2.8 and 20.5 degrees C, respectively, with only a minor reduction in fiber stiffness. The maximum speed of shortening was decreased by lowered pH by 12.9 +/- 1.5 and 7.8 +/- 1.1% at low and high temperature, respectively. Acidification increased the time to reach 70% of maximum force by 18.0% at approximately 2 degrees C; the same pH change performed at approximately 20 degrees C in the same fibers reduced the rise time by 24.1%. The same increase in the rate of rise of force at high temperature was also found at normal pH after the fibers were fatigued by frequent stimulation. It is concluded that, in frog muscle, the force-depressant effect of acidification does not vary significantly with temperature. By contrast, acidification affects the onset of activation in a manner that is critically dependent on temperature.

A mechanical stretch induces contractile activation in unstimulated developing rat skeletal muscle in vitro.

The effects of a stretch-release cycle (approximately 25% of the resting muscle fibre length, Lo) on both tension and [Ca2+]i in small, unstimulated, intact muscle fibre bundles isolated from adult and neonatal rats were investigated at 20 degrees C. The results show that the effects of the length change depended on the age of the rats. Thus, the length change produced three effects in the neonatal rat muscle fibre bundles, but only a single effect in the adult ones. In the neonatal fibre bundles, the length change led to an increase in resting muscle tension and to a transient increase in [Ca2+]i. The stretch-release cycle was then followed by a twitch-like tension response. In the adult fibre bundles, only the increase in resting tension was seen and both the transient increase in [Ca2+]i and the stretch-induced twitch-like tension response were absent. The amplitude of the twitch-like tension response was affected by both 2,3-butanedione monoxime and sarcomere length in the same manner as active twitch tension, suggesting that it arose from actively cycling crossbridges. It was also reversibly abolished by 25 mM K+, 1 microM tetrodotoxin and 1.5 mM lidocaine (lignocaine), and was significantly depressed (P < 0.001) by lowering [Ca2+]o. These findings suggest that a rapid stretch in neonatal rats induces a propagated impulse that leads to an increase in [Ca2+]i, and that abolishing the action potential abolishes the stretch-induced twitch-like tension response. In 5- to 7-day-old rats, the twitch-like tension response was approximately 50 % of the isometric twitch. It then decreased progressively with age and was virtually absent by the time the rats were 21 days old. Interestingly, this is the same period over which rat muscles differentiate from their neonatal to their adult types.

Contractile properties of isolated muscle spindles of the frog.

Force and isotonic shortening velocities were studied (0.6-4.0 degrees C) in isolated single muscle spindles from the anterior tibialis muscle of Rana temporaria using techniques that enabled measurements both from the spindle as a whole and from marked segments of the preparation. The force-velocity relationship during tetanic stimulation exhibited the same biphasic shape as previously described for extrafusal muscle fibres. However, the maximum speed of shortening of the spindle fibres was merely 0.95 +/- 0.006 lengths s(-1) (mean +/- S.E.M., n = 11), which is approximately half the value recorded in extrafusal fibres of the same muscle. The maximum tetanic force, 91 +/- 10 kN m(-2), n = 14, was likewise only approximately half that produced by extrafusal fibres. The force generated by the capsule segment was lower than that produced by the whole spindle resulting in elongation of the capsule region during a fixed-end tetanus. The intracellular calcium ion concentration reached during the plateau of the tetanus, 1.7 +/- 0.1 microM (n = 8), was substantially lower than the value attained in extrafusal fibres under equivalent conditions. In accordance, the spindle fibres did not become fully activated during supramaximal electrical stimulation as indicated by the finding that the tetanic force could be further increased by 16.6 +/- 0.04 % (n = 5) on addition of 0.5 mM caffeine. Inadequate activation may thus, to a certain extent, account for the relatively low force per cross-sectional area of the spindle fibres. The contractile properties of the intrafusal fibres should make the spindle organ suited to provide feedback control during eccentric (forced lengthening) and static (isometric) contractions and, with reduced effectiveness, during slow muscle shortening.