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Dynamic stiffness and crossbridge action in muscle.

Small sinusoidal vibrations at 300 HZ were applied to frog sartorius muscle to measure the dynamic stiffness (Young's modulus) throughout the course of tetanus. For a peak-to-peak amplitude of 0.4% the dynamic Young's modulus increased from 1.5 X 10(5) Nm-2 in the resting state to 2 X 10(7) Nm-2 in tetanus. After correction for the external connective tissue, the dynamic Young's modulus of the muscle was almost directly proportional to the tension throughout the development of tetanus. The ratio of dynamic Young's modulus to tensile stress thus remained constant (with a value at 300 Hz of approximately 100), consistently with Huxley and Simmon's identification of the crossbridges as the source of both tension and stiffness. For a single crossbridge the ratio of stiffness to tension was 8.2 X 10(7) m-1 at 300 Hz; it is deduced from literature data that the limiting value at high frequencies is about 1.6 X 10(8) m-1. This ratio is interpreted on Harrington's (1971) model to show that crossbridge action can be explained by a helix-coil transition of about 80 out of the 260 residues in each S-2 myosin strand. It is also shown that a helix-coil model can account for the observed rapid relaxation of muscle without invoking any complex behaviour of the crossbridge head.

Animals↗

Changes in the properties of synaptic channels opened by acetylcholine in denervated frog muscle.

Acetylcholine (ACh)-activated channels in end-plates of frog sartorius muscle were studied at various times after denervation. Mean open times of the synaptic membrane channels were derived from the time constant of decay of miniature end-plate currents (tau MEPC) evoked by ACh quanta released from Schwann cells, which replace the motor nerve terminals after these degenerate. Membrane current noise, elicited by iontophoretic application of ACh to voltage-clamped end-plates, was also used to determine mean open time (tau noise) and conductance of the ion channels. About 1 week after denervation, soon after Schwann cell MEPCs appeared, they had a tau similar to that of the neural MEPC in innervated end-plates. However, 5-6 weeks after denervation tau MEPC was increased by a factor of about 5. Circa 4 weeks after denervation, cholinesterase activity of the denervated muscle decreased to 76% of that in the contralateral, innervated muscle, and even 4 months after the operation it was still 64%. Thus, it is unlikely that a change in acetylcholinesterase activity is the main factor responsible for the increase in tau of Schwann cell MEPC. About 1 week after denervation tau noise was close to that in innervated end-plates (about 2 ms). Twelve to 24 days after denervation the average channel open time was 4.5 +/- 1.0 ms, with some end-plates still showing normal 'fast' channels. However, in muscles denervated for 47-113 days the open time was 12.9 +/- 1.9 ms. In the early and intermediate periods, ACh-induced noise spectra with two components were obtained from many end-plates, indicating the simultaneous activation of two different types of channels. At some end-plates during the early and intermediate periods after denervation, but not after about 5 weeks, neostigmine caused the appearance of a component, which was as fast as that of normal end-plate channels. In other experiments small doses of alpha-bungarotoxin were applied in order to predominantly block extra-junctional receptors. In the early period of denervation, when two components were present in the noise spectra, alpha-bungarotoxin eliminated the slow component leaving channels as fast as in innervated end-plates. After prolonged denervation, a component with tau of about 5.5 ms was occasionally disclosed by application of alpha-bungarotoxin. tau noise and tau MEPC from the same end-plate closely agreed. Our results indicate that at frog end-plates the open time of the majority of the synaptic channels opened by ACh becomes longer with increasing time after denervation.

Acetylcholine↗

Effect of force development on contraction induced glucose transport in fast twitch rat muscle.

A previous study has shown that in fast twitch frog sartorius muscle contraction stimulated glucose transport depends only on stimulation frequency and not on workload. In contrast, we have recently shown that in rat slow twitch muscle stimulated to contract at constant frequency, glucose transport varies directly with force development and, in turn, metabolism. The present study was carried out to clarify whether the discrepancy between the earlier studies reflected differences in physiological behaviour between fast and slow twitch muscle. We investigated the effect of force development on glucose transport in incubated fast twitch rat flexor digitorum brevis (rich in type 2a fibres) and epitrochlearis (rich in type 2b fibres) muscle. Muscles were electrically stimulated to perform repeated tetanic contractions at 1 Hz for 10 min. Resting length was adjusted to achieve either no force or maximum force. Glucose transport (2-deoxyglucose uptake) increased when force was produced compared with when it was not (P < 0.05) in both flexor digitorum brevis (19 +/- 7 (basal), 163 +/- 14 (no force) and 242 +/- 17 (max force) nmol x g(-1) x 5 min(-1)) and epitrochlearis (60 +/- 4 (basal), 100 +/- 7 (no force) and 125 +/- 6 (max force) nmol x g(-1) x 5 min(-1)). In both muscles glucose transport increased in parallel with metabolic rate, as reflected by muscle lactate concentrations and 5' AMP-activated protein kinase activity, during contractions. In conclusion, as previously shown for rat soleus muscle, at a given stimulation frequency glucose transport varies directly with force development in rat flexor digitorum brevis and epitrochlearis muscle. Accordingly, force development enhances glucose transport in all mammalian muscle fibre types. The influence of force development probably reflects effects of enhanced 5' AMP-activated protein kinase activity resulting from reduced intra-cellular energy status and pH.

AMP-Activated Protein Kinase Kinases↗

The penetration of some cations into muscle.

The influx of Na(+), K(+), Rb(+), and Cs(+) into frog sartorius muscle has been followed. The results show that a maximum rate is found for K(+), while Na(+) and Cs(+) penetrate much more slowly. Similar measurements with Ca(++), Ba(++), and Ra(++) show that Ba(++) penetrates at a rate somewhat greater than that of either Ca(++) or Ra(++). All these divalent cations, however, penetrate at rates much slower than do the alkali cations. The results obtained are discussed with reference to a model that has been developed to explain the different penetration rates for the alkali cations.

Cations↗

Multimodal distribution of frog miniature endplate potentials in adult denervated and tadpole leg muscle.

Amplitude histograms of spontaneous miniature endplate potentials (MEPPs) from adult sartorius muscle cells show a definite bimodality with the mean amplitude of the larger mode five to seven times that of the smaller mode which accounted for 2-5 % of the total MEPPs. Histograms were plotted after high frequency MEPP generation induced by increasing temperature, increasing external calcium or nerve stimulation. These plots showed a reversible left-shift of the major mode as well as a reversible increase in the proportion of small mode MEPPs. Repeated challenges shifted almost all MEPPs into the small mode. An increase in the percentage of small mode MEPPs also occurred spontaneously during the course of denervation before the quiescent period and some of the histogram profiles showed multiple modes whose means were integer multiples of the small mode mean. In the early stages of hind leg development the greatest proportion of MEPPs were of the small mode size; as metamorphosis progressed, the histograms showed a definite multimodality with the mean of each mode being an integer multiple of the small mode mean and with the proportion of MEPPs in each mode about the same. During tail resorption the percentage of larger MEPPs increased until the adult histogram profile was reached. Thus, the changes in MEPP amplitude histograms over the course of metamorphosis are the reverse of those found with denervation.

Animals↗

Sodium currents in skeletal muscle fibres from the toad Bufo marinus.

Single fibres from the sartorius muscle of the toad Bufo marinus were used to study ionic currents, using a fast voltage-clamp method. Sodium currents were analysed following Hodgkin-Huxley formulations. Internal caesium fluoride blocks delayed currents but produced shifts in the equilibrium potential of the early channel due to sodium accumulation. Delayed channels are very unstable when fluoride is used inside the fibre. These instabilities and equilibrium potential shifts were eliminated on replacing fluoride by aspartate in the internal solution. Late peaks of inward current, probably associated with activity at the tubular system, were occasionally observed for small depolarizations. The results are compared with those obtained for other amphibian species and used to explain more general electrophysiological properties of muscle fibres of this tropical toad.

Animals↗

Metachromatic procedure for fiber typing in chicken skeletal muscle.

Metachromatic staining of anterior latissimus dorsi, posterior latissimus dorsi, and sartorius muscles of domestic chicken with azure A and toluidine blue produced differentiation of muscle fiber types. The procedure is rapid, with optimum results obtained after 2 to 5 min incubation and 5 to 10 s of staining. Reaction of serial sections for myosin Ca++-adenosine triphosphatase with the usual procedure showed that the same fibers were typed similarly with both methods.

Animals↗

[Effect of insulin and adrenaline on the respiration and glycolysis of the skeletal muscle of frog].

It was shown in the experiments on isolated sartorius muscles of the frog that insulin increased endogenous respiration by 10 mm3 O2 hr per gramm of wet weight. The effect of insulin and epinephrine on respiration is additive. Insulin in middle (5--50 milliunits/ml) and low, physiological, (50 microunits/ml) concentration does not induce acceleration of glycolyses. The conclusion was confirmed that the increase of the endogenous respiration by insulin cannot be accounted for by the mobilization of the substrate of glycolysis. In the presence of epinephrine, the low concentration (50 microunits/ml) of insulin induces the decrease of the content of glucose-6-phosphate. It is probably connected with the influence of insulin on the concentration level of the cAMP.

Adenosine Triphosphate↗

The distribution of orthogonal assemblies and other intercalated particles in frog sartorius and rabbit sacrospinalis muscle.

Freeze-fracture replicas have been prepared from two fast-acting vertebrate muscles (frog sartorius and rabbit sacrospinalis) and are described with particular reference to the distribution of intercalated particles in the plasma membrane. T-tubule and SR cisternae. Orthogonal assemblies of small particles are present on the A face plasma membrane in each instance, and their distribution (in sartorius) is found to be random with respect to the underlying myofibrillar sarcomere repeat. Such assemblies are not present on A or B faces of T-tubule or SR cisternae. Asymmetric particle distribution is described for fracture faces of the T-tubules and SR: the profuse particle packing of the SR A face is uniform from terminal cisternae to medial fenestrated collar. Intercalated particles are present on A and B faces of T-tubule fractures; more commonly on the latter. These results are compared with studies on insect muscles, and a comparative approach to further studies on the correlation between membrane structure and function is discussed.

Animals↗

Organization of single motor units in feline sartorius.

1. We depleted single motor units in feline sartorius muscles of glycogen by stimulating their motoneurons intracellularly. We mapped the intramuscular distribution of depleted fibers by inspecting histological cross-sections throughout the length of sartorius. 2. We selected ten depleted motor units for detailed study and quantitative analysis. Nine motor units were located in the anterior head of sartorius. One was located in a muscle whose distal half appeared to have been damaged some time before the acute experiment. A single motor unit was located in the medial head of sartorius. 3. Five motor units were composed of fast-twitch glycolytic (FG) muscle fibers, two of fast-twitch oxidative glycolytic (FOG) muscle fibers, and three of slow-twitch oxidative (SO) muscle fibers. Estimates of the numbers of depleted fibers in motor units of anterior sartorius indicated that FG motor units were larger (mean 566 fibers) than FOG and SO motor units (SO mean 190, FOG mean 156 fibers). The SO motor unit in the damaged muscle had 550 fibers. One motor unit depleted in the medial head of sartorius had 270 fibers with FG profiles. 4. Muscle fibers belonging to each anterior motor unit were never distributed throughout the whole cross-section of anterior sartorius at any proximodistal level. Furthermore, fibers were distributed nonuniformly along the proximodistal axis of the muscle. In most muscles at least a few depleted fibers were found at all proximodistal levels. However, in one normal muscle and the damaged muscle, depleted fibers were confined to the proximal end. 5. The fibers in the medial motor unit were confined to a strip that did not extend across the whole cross-section of the muscle head. Fibers within this strip were scattered quite evenly from origin to insertion. This medial FG motor unit occupied a smaller territory and contained fewer fibers than anterior motor units of the same histochemical type. 6. These results show that sartorius motor units are not distributed uniformly in the mediolateral plane; those in anterior sartorius were distributed asymmetrically in the proximodistal axis as well. This finding has important functional implications for the way in which we model force development and transmission in sartorius and other long muscles.

Animals↗

Modelling diffusive O(2) supply to isolated preparations of mammalian skeletal and cardiac muscle.

The purpose of this study was to use A. V. Hill's equation describing diffusion of O(2) into cylindrical muscles to assess the adequacy of O(2) supply for commonly used isolated preparations of mammalian cardiac and skeletal muscles. The diffusion equation was solved numerically to give the maximum, steady state O(2) diffusion distances (i.e. the distance from the surface of the muscle to the radial location where P(O(2)) is 0) for both resting and contracting muscles and for a range of temperatures. Non-steady state solutions for the rest-to-work transition were also determined to estimate how long contractile activity could be continued before anoxia develops at the muscle centre. The influence on muscle oxygenation of myoglobin-facilitated O(2) diffusion was also assessed. The analysis was performed for typical sized, whole muscles from adult rats and mice, for frog sartorius muscle and for a range of temperatures. Muscle O(2) consumption rates were taken from the literature. The results indicated that (1) diffusive O(2) supply would be adequate to support resting metabolism of soleus and EDL muscles of rat and mouse but may not be adequate to support the transient high resting metabolic rate of papillary muscles shortly after dissection, (2) during steady contractile activity of soleus and EDL muscles, particularly those from the rat, over a reasonable range of duty cycles, adequate O(2) supply could only be ensured if the radii of preparations was substantially smaller than those of whole muscles and (3) for cardiac muscles, diffusive O(2) supply could only support steady-state metabolism at twitch frequencies <1 Hz for whole papillary muscles from rat and <3 Hz for those from mouse. Reducing experimental temperature markedly enhances O(2) supply to skeletal, but not cardiac, muscle. O(2) supply from myoglobin had only minimal effects on oxygenation under typical isolated muscle conditions.

Animals↗

Selective loss of a plasma membrane protein associated with contraction of skeletal muscle.

Muscle proteins were labeled by incubating isolated frog sartorius muscles with [3H]- or [14C]phenylalanine. Sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis of plasma membrane fractions revealed a major protein band with an apparent molecular weight of approx. 96,000. Radioactivity in this band showed a clearly delineated decrease, relative to other bands, when previously labeled muscles were induced to contract either by electrical stimulation or by increasing the influx of Ca2+ from the incubation medium. It is postulated that a Ca2+-activated neutral protease may account for this decrease in labeled membrane protein.

Animals↗

Static and dynamic x-ray diffraction recordings from living mammalian and amphibian skeletal muscles.

Static and time-resolved two-dimensional x-ray diffraction patterns, recorded from the living mouse diaphragm muscle, were compared with those from living frog sartorius muscle. The resting pattern of mouse muscle was similar to that of frog muscle, and consisted of actin- and myosin-based reflections with spacings basically identical to those of frog. As a notable exception, the sampling pattern of the myosin layer lines (MLL's) indicated that the mouse myofilaments were not organized into a superlattice as in frog. The intensity changes of reflections upon activation were also similar. The MLL's of both muscles were markedly weakened. Stereospecific (rigorlike) actomyosin species were not significantly populated in either muscle, as was evidenced by the 6th actin layer line (ALL), which was substantially enhanced but without a shift in its peak position or a concomitant rise of lower order ALL's. On close examination of the mouse pattern, however, a few lower order ALL's were found to rise, slightly but definitely, at the position expected for stereospecific binding. Their quick rise after the onset of stimulation indicates that this stereospecific complex is generated in the process of normal contraction. However, their rise is still too small to account for the marked enhancement of the 6th ALL, which is better explained by a myosin-induced structural change of actin. Since the forces of the two muscles are comparable regardless of the amount of stereospecific complex, it would be natural to consider that most of the force of skeletal muscle is supported by nonstereospecific actomyosin species.

Actins↗

Effects of chronic heat exposure and protein intake on growth performance, nitrogen retention and muscle development in broiler chickens.

The respective effects of ambient temperature, dietary crude protein and feed intake were investigated in finishing chickens and the consequence of protein supplementation under high temperature conditions was analysed in particular. Heat-related reduction in growth was associated with decreased nitrogen retention (-30 or -35% according to the diet), which could not be explained by the observed lower feed intake alone. Tissue samples performed in 5- to 6-week-old chicks showed varying effects of heat according to the muscles studied: at 32 degrees C, the proportion of Pectoralis major muscle (in percentage of body weight) appeared slightly reduced (reduction lower than 10%), whereas the proportion of two leg muscles were increased (+10 to +15% for the Sartorius muscle; +5% for the gastrocnemius muscle). At 32 degrees C, providing a high protein diet significantly (P < 0.05) increased weight gain and feed efficiency, and slightly improved whole body protein deposition.

Animal Nutritional Physiological Phenomena↗

Work and power output in the hindlimb muscles of Cuban tree frogs Osteopilus septentrionalis during jumping.

It has been suggested that small frogs use a catapult mechanism to amplify muscle power production during the takeoff phase of jumping. This conclusion was based on an apparent discrepancy between the power available from the hindlimb muscles and that required during takeoff. The present study provides integrated data on muscle contractile properties, morphology and jumping performance that support this conclusion. We show here that the predicted power output during takeoff in Cuban tree frogs Osteopilus septentrionalis exceeds that available from the muscles by at least sevenfold. We consider the sartorius muscle as representative of the bulk of the hindlimb muscles of these animals, because this muscle has properties typical of other hindlimb muscles of small frogs. At 25 degrees C, this muscle has a maximum shortening velocity (Vmax) of 8.77 +/- 0.62 L0 s-1 (where L0 is the muscle length yielding maximum isometric force), a maximum isometric force (P0) of 24.1 +/- 2.3 N cm-2 and a maximum isotonic power output of 230 +/- 9.2 W kg-1 of muscle (mean +/- S.E.M.). In contrast, the power required to accelerate the animal in the longest jumps measured (approximately 1.4 m) is more than 800 W kg-1 of total hindlimb muscle. The peak instantaneous power is expected to be twice this value. These estimates are probably conservative because the muscles that probably power jumping make up only 85% of the total hindlimb muscle mass. The total mechanical work required of the muscles is high (up to 60 J kg-1), but is within the work capacities predicted for vertebrate skeletal muscle. Clearly, a substantial portion of this work must be performed and stored prior to takeoff to account for the high power output during jumping. Interestingly, muscle work output during jumping is temperature-dependent, with greater work being produced at higher temperatures. The thermal dependence of work does not follow from simple muscle properties and instead must reflect the interaction between these properties and the other components of the skeletomuscular system during the propulsive phase of the jump.

Animals↗

[Effects of estrone on transmission at neuromuscular junction of the toad].

The effects of estrone on neuromuscular transmission were studied intracellularly at the muscle fiber endplate in isolated sciatic nerve-sartorius muscle preparations of the toad (Bufo bufo gargarizans) in vitro. The results showed that estrone (10(-4) mol/L), added to bath medium, caused an increase in amplitudes of endplate potentials (EPP) both in 10 micrograms tubocurarine treated and 10 mmol/L MgCl2 treated preparations, but the increase was more prominent in the former, and that estrone, 30 min after being added to bath medium, caused clear increase both in frequency and amplitude of miniature endplate potentials (MEPP), but the wave form of MEPP remained unchanged and the resting membrane potentials were also unchanged. These results suggest that estrone can increase neuromuscular transmission, which seems to involve primarily the presynaptic mechanism.

Animals↗

The thermoelastic effect in rigor muscle of the frog.

Small length changes were imposed on pairs of sartorius muscles from Rana temporaria and Rana pipiens in rigor and the mechanical and thermal responses studied. Rigor was induced by soaking the muscles overnight at 0 degrees C in a physiological salt solution containing 1.5 mM sodium azide and 0.4 mM sodium iodoacetate. Tension was measured at both the tibial and the pelvic ends of the preparation. Muscles were held at a steady tension of 20 to 76 kN m-2 and stretches or releases of 0.02 to 0.6 mm applied in pairs, with the initial change reversed several hundred milliseconds later. Single stretches resulted in heat absorption and releases in heat production by the preparation. Net heat production resulted from complete cycles of length changes larger than 0.1 mm, whether the initial change was a stretch or a release. The heat produced by the complete cycle was attributed to the movement of the muscles over the thermopile. It was proportional to the difference in tension between the tibial and pelvic ends of the preparation and increased with the size and speed of the length change. Half the heat produced by a complete cycle of length changes was subtracted from the thermal response recorded in the first half-cycle to obtain the reversible component of the response. The reversible component was linearly related to the tension change for all sizes and speeds of length change which were studied, with the heat:tension ratio ranging from -0.0093 to -0.0179 in eleven muscles (mean -0.0128 +/- 0.0009). The constancy of the heat:tension ratio in rigor muscles over a wide range of mechanical conditions indicates that the source of the thermal changes is the normal elasticity of the preparation. Since the size of the ratio is approximately the same as that measured in active muscles, the tension-dependent component of the thermal response to length changes applied to active muscles is probably also of elastic origin.

Animals↗