Ultrastructural diversity in motor units of crustacean stomach muscles

The physiological and ultrastructural properties of muscle fiber.s comprising three motor units in the gastric mill of blue crabs are described. In their contractile properties muscle fibers in all motor units are similar and resemble the slow type fibers in crustacean limb muscles. The majority of fibers generate large excitatory post-synaptic potentials which do not facilitate strongly. Structurally two types of fibers are found. The one type has long sarcomeres (greater than 6 mum), thin to thick myofilament ratios of 5-6:1 and diads located near the ends of the A-band. The other type has shorter sarcomeres (less than 6 mum), thin to thick myofilament ratios of 3:1 and diads located at mid sarcomere level. Both types of fibers occur within a single motor unit and this differs from the vertebrate situation. Furthermore, the finding of fibers with a low thin to thick myofilament ratio of 3:1 demonstrates that they are not exclusive to fast type crustacean muscle but also occur in slow stomach muscles.     

Summation of motor unit forces in rat medial gastrocnemius muscle

The summation of contractile forces of motor units (MUs) was analyzed by comparing the recorded force during parallel stimulation of two and four individual MUs or four groups of MUs to the algebraic sum of their individual forces. Contractions of functionally-isolated single MUs of the medial gastrocnemius muscle were evoked by electrical stimulation of thin filaments of the split L5 or L4 ventral roots of spinal nerves. Additionally, contractions of large groups of MUs were evoked by stimuli delivered to four parts of the divided L5 ventral root. Single twitches, 40 Hz unfused tetani, and 150 Hz fused maximum tetani were recorded. In these experimental situations the summation was more effective for unfused tetani than for twitches or maximum tetani. The results obtained for pairs of MUs were highly variable (more- or less-than-linear summation), but coactivation of more units led to progressively weaker effects of summation, which were usually less-than-linear in comparison to the algebraic sums of the individual forces. The variability of the results highlights the importance of the structure of the muscle and the architecture of its MUs. Moreover, the simultaneous activity of fast and slow MUs was considerably more effective than that of two fast units.     

Organization of skeletal muscle in the urodele Triturus cristatus: muscle fibre types and motor units

Four muscle fibre types are described in the biceps and extensor digitorum communis muscles of the newt forelimb. The histological criteria forming the basis for the distinctions include differential staining with p-phenylenediamine and succinate dehydrogenase histochemistry and electron microscopy. In addition, three distinctive motor unit types are described for the biceps muscle. These are fast units, slow units and intermediate units. The structure of muscle fibre and the physiological characteristics of muscle fibres belonging to each motor unit, have been correlated by using iontophoretic passage of Lucifer yellow into muscle fibres belonging to physiologically characterized motor units and their subsequent histological identification by the succinate dehydrogenase reaction. The three motor unit types correspond to slow muscle fibres, intermediate muscle fibres and two classes of fast muscle fibres.     

Interspecies differences of motor units properties in the medial gastrocnemius muscle of cat and rat

The purpose of the study was to analyze the interspecies differences of motor unit contractile properties in two most frequently studied mammals: cats and rats. A total sample of 166 motor units (79 in cats and 85 in rats) was investigated in the medial gastrocnemius muscle. Considerable differences were found in composition of the studied muscle. In cats, fast fatigable, fast resistant and slow units formed 68, 18 and 14% of the investigated population, whereas in rats 36, 52 and 12%, respectively. The contraction and relaxation times of motor units in the cat muscle were evidently longer than in the rat and the border values for fast/slow motor units division in these species were 44 and 20 ms, respectively. The mean values of twitch and tetanic forces appeared to be 7-8 times lower in rats, for fast, while 2-5 times for slow motor units. Also variability between the strongest and the weakest units within each type revealed differences 10-60 times in cats, whereas only 3.5-14 times in rats. The summation of twitches into tetanus for fast units was comparable in both species, but for S units was evidently more effective in the cat. In fast motor units' tetanic contractions evident interspecies differences concerned sag appearance and profiles of unfused tetani of FF and FR units. Differences in contractile properties described in the study may depend on the size, number and innervation ratio of motor units in the muscle of cat and rat, as well as their biochemical variability. Differences in composition of motor unit types and uneven mechanisms of force development may reflect biological adaptation to variable behaviour of cats and rats.     

Comparative analysis of motor unit action potentials of the medial gastrocnemius muscle in cats and rats

Properties of motor unit action potentials (MUAPs) were compared for medial gastrocnemius (MG) motor units (MUs) in cats and rats. The experiments on functionally isolated MUs were performed under general anaesthesia, under comparable conditions (surgery, stimulating protocol and recording methods) for both species investigated. The proportions of motor units and contractile properties of the sample used in the study were consistent with previous studies performed on the MG muscle in both animal species, so comparisons of action potentials of individual types of MUs were acknowledged as fully reliable. The most prominent differences concerning MUAPs were observed in total duration and peak-to-peak times which for all MU types were about twice longer in cat MUs, in comparison to the rat MUs. The considerable disproportions were observed between the MUAP amplitudes of FF (fast fatigable), FR (fast resistant to fatigue) and S (slow) MUs in each species (the highest amplitudes were measured for FF and the lowest for S MUs), but there were no significant differences between cat and rat when respective types of MUs were compared. The shapes of MUAPs were commonly characterized by biphasic waveforms composed of two or three turns in all types of units, and no interspecies differences were revealed. Several factors influencing MUAP parameters were discussed indicating most of all importance of variable length of cat and rat muscle fibres and ambiguous influence of motor unit size, thickness of muscle fibres and their density around the recording electrode in the MG muscle of both species.     

Myosin isozymes in normal and cross-reinnervated cat skeletal muscle fibers

Immunocytochemical characteristics of myosin have been demonstrated directly in normal and cross-reinnervated skeletal muscle fibers whose physiological properties have been defined. Fibers belonging to individual motor units were identified by the glycogen-depletion method, which permits correlation of cytochemical and physiological data on the same fibers. The normal flexor digitorum longus (FDL) of the cat is composed primarily of fast-twitch motor units having muscle fibers with high myosin ATPase activity. These fibers reacted with antibodies specific for the two light chains characteristic of fast myosin, but not with antibodies against slow myosin. Two categories of fast fibers, corresponding to two physiological motor unit types (FF and FR), differed in their immunochemical response, from which it can be concluded that their myosins are distinctive. The soleus (SOL) consists almost entirely of slow-twitch motor units having muscle fibers with low myosin ATPase activity. These fibers reacted with antibodies against slow myosin, but not with antibodies specific for fast myosin. When the FDL muscle was cross-reinnervated by the SOL nerve, twitch contraction times were slowed about twofold, and motor units resembled SOL units in a number of physiological properties. The corresponding muscle fibers had low ATPase activity, and they reacted with antibodies against slow myosin only. The myosin of individual cross-reinnervated FDL muscle units was therefore transformed, apparently completely, to a slow type. In contrast, cross-reinnervation of the SOL muscle by FDL motoneurons did not effect a complete converse transformation. Although cross-reinnervated SOL motor units had faster than normal twitch contraction times (about twofold), other physiological properties characteristic of type S motor units were unchanged. Despite the change in contraction times, cross-reinnervated SOL muscle fibers exhibited no change in ATPase activity. They also continued to react with antibodies against slow myosin, but in contrast to the normal SOL, they now showed a positive response to an antibody specific for one of the light chains of fast myosin. The myosins of both fast and slow muscles were thus converted by cross-reinnervation, but in the SOL, the newly synthesized myosin was not equivalent to that normally present in either the FDL or SOL. This suggests that, in the SOL, alteration of the nerve supply and the associated dynamic activity pattern are not sufficient to completely respecify the type of myosin expressed.     

Differences between properties of male and female motor units in the rat medial gastrocnemius muscle.

Differences between motor units in hindlimb locomotor muscles of male and female Wistar rats were studied. The contractile and action potential properties of various types of motor units as well as proportions of these units in the medial gastrocnemius muscle were analyzed. Experiments were based on functional isolation and electrical stimulation of axons of single motor units. Composition of motor units was different for male and female subjects, with higher number of the fast fatigable and lower number of slow type units in male animals. The contraction and the half-relaxation times were significantly longer in male motor units, what might be due to differences in muscle size. Slower contraction of male motor units likely corresponds to lower firing rates of their motoneurons. On the other hand, no significant differences between sexes were observed with respect to force parameters of motor units (the twitch and the maximum tetanus forces), except the fast resistant units (higher force values in male muscles). The mass of the muscle was approximately 1.5 time bigger in male rats. However, the mean ratio of motor unit tetanus force to the muscle mass was almost twice smaller in this group, what indirectly suggests that muscles of male rats are composed of higher number of motor units. Finally, female muscles appeared to have higher fatigue resistance as the effect of higher proportion of resistant units (slow and fast resistant) and higher values of the fatigue index in respective motor unit types. The motor unit action potentials in female rats had slightly lower amplitudes and shorter time parameters although this difference was significant only for fast resistant units.     

Restoration of muscle contractile properties after partial and complete denervation

The subject of this experimental study was the isometric contractile properties of rat tibialis anterior muscle, number and average size of the motor units as well as type content and type-grouping of muscle fibres according to SDH activity in the same muscle after total and partial denervation (crushing the sciatic nerve and L4). It has been shown that in the process of reinnervation after total and partial denervation, quantitative differences with the general tendency in the dynamics of restoration of contractile properties of the whole muscle are found at different dynamics of restoration of electromyographic and muscle histochemical characteristics of motor units.     

Time-related changes of motor unit properties in the rat medial gastrocnemius muscle after spinal cord injury. I. Effects of total spinal cord transection

The contractile properties of motor units (MUs) were electrophysiologically investigated in the medial gastrocnemius (MG) muscle in 17 Wistar three-month-old female rats: 14, 30, 90 and 180 days after the total transection of the thoracic spinal cord and compared to those in intact (control) rats. A sag phenomenon, regularly observed in unfused tetani of fast units in intact animals at 40 Hz stimulation, almost completely disappeared in spinal rats. Therefore, the MUs of intact and spinal rats were classified as fast or slow types basing on 20 Hz tetanus index, the value of which was lower or equal 2.0 for fast and higher than 2.0 for slow MUs. The MUs composition of MG muscle changed with time after the spinal cord transection: an increasing proportion of fast fatigable (FF) units starting one month after injury and a disappearance of slow (S) units within the three months were observed. In all MUs investigated the twitch contraction and half-relaxation time were significantly prolonged after injury (p < 0.01, Mann–Whitney U-test). Moreover, a decrease of the fatigue index for fast resistant (FR) and slow MUs was observed in subsequent groups of spinal rats. No significant changes were found between twitch forces in all MU types of spinal animals (p > 0.05). However, due to a decrease of the maximal tetanic force, a significant rise of the twitch-to-tetanus ratio of all MUs in spinal rats was detected (p < 0.01). The considerable reduction of ability to potentiate the force was noticed for fast, especially FF type MUs. In conclusion, the spinal cord transection leads to changes in the proportion of the three MU types in rat MG muscle. The majority of changes in MUs’ contractile properties were developed progressively with time after the spinal cord injury. However, the most intensive alterations of twitch-time parameters were observed in rats one month after the transection.     

Power and control muscles of cicada song: structural and contractile heterogeneity

Sound production in cicadas is powered by a pair of large muscles whose contractions cause buckling of cuticular tymbals and thereby create sound pulses. Sound is modulated by control muscles that alter the stiffness of the tymbals or change the shape of the abdominal resonance chamber. Muscle ultrastructure and contractile properties were characterized for the tymbal muscle and two control muscles, the ventral longitudinal muscle and the tymbal tensor, of the periodical cicada Magicicada septendecim. The tymbal muscle is a fast muscle that is innervated by a single motoraxon. The control muscles are an order of magnitude less massive than the tymbal muscles, but their innervation patterns were considerably more complex. The tensor muscle is innervated by two axons, each of which evokes rather slow twitches, and the ventral muscle is innervated by at least six axons, some of which produce fast and the others slow contractions. Muscle contraction kinetics correlated well with ultrastructure. Fibers of the tymbal muscle and the portions of the ventral muscle thought to be fast were richly supplied with transverse tubules (T-tubules) and sarcoplasmic reticulum (SR); slow portions of the ventral muscle and the tensor muscle had relatively little SR.Abbreviations SR sarcoplasmic reticulum - TTS transverse tubular system - VLM ventral longitudinal muscle     

Characteristics of surface mechanomyogram are dependent on development of fusion of motor units in humans.

The purpose of this study was to test whether surface mechanomyogram (MMG) recorded on the skin reflects the contractile properties of individual motor units in humans. Eight motor units in the medial gastrocnemius muscle were identified, and trains of stimulation at 5, 10, 15, and 20 Hz were delivered to each isolated motor unit. There was a significant positive correlation between the duration of MMG and twitch duration. MMG amplitude decreased with increasing stimulation frequency. Reductions in MMG amplitude were in parallel with the reductions in force fluctuations, and the rate of change in both was positively correlated across the motor units. Rate of change in MMG amplitude against force was negatively correlated to half relaxation time and twitch duration. Similar negative correlations were found between force fluctuations and contractile properties. These results provide evidence supporting a direct relation between MMG and contractile properties of individual motor units within the gastrocnemius muscle, indicating that surface MMG is dependent on the contractile properties of the activated motor units in humans.     

The oculomotor system of Daphnia magna

Summary The highly mobile cyclopic compound eye of Daphnia magna is rotated by six muscles arranged as three bilateral pairs. The three muscles on each side of the head share a common origin on the carapace and insert dorsally, laterally and ventrally on the eye. The dorsal and ventral muscles are each composed of two muscle fibers and the lateral muscle is composed of from two to five fibers, with three the most common number. Individual muscle fibers are spindle-shaped mononucleated cells with organized bundles of myofilaments. Lateral eye-muscle fibers are thinner than those of the other muscles but are otherwise similar in ultrastructure. Two motor neurons innervate each dorsal and each ventral muscle and one motor neuron innervates each lateral muscle. The cell bodies of the motor neurons are situated dorsally in the supraesophageal ganglion (SEG) and are ipsilateral to the muscles they innervate. The dendritic fields of the dorsal-muscle motor neurons are ipsilateral to their cell bodies; those of the ventral-muscle motor neurons are bilateral though predominantly contralateral. The central projections of the lateral-muscle motor neurons are unknown. In the dorsal and ventral muscles one motor axon synapses principally with one muscle fiber; in each lateral muscle the single motor axon branches to, and forms synapses with, all the fibers. The neuromuscular junctions, characterized by pre- and postsynaptic densities and clear vesicles, are similar in all the eye muscles.     

The resilience of the size principle in the organization of motor unit properties in normal and reinnervated adult skeletal muscles

Henneman's size principle relates the input and output properties of motoneurons and their muscle fibers to size and is the basis for size-ordered activation or recruitment of motor units during movement. After nerve injury and surgical repair, the relationship between motoneuron size and the number and size of the muscle fibers that the motoneuron reinnervates is initially lost but returns with time, irrespective of whether the muscles are self- or cross-reinnervated by the regenerated axons. Although the return of the size relationships was initially attributed to the recovery of the cross-sectional area of the reinnervated muscle fibers and their force per fiber, direct enumeration of the innervation ratio and the number of muscle fibers per motoneuron demonstrated that a size-dependent branching of axons accounts for the size relationships in normal muscle, as suggested by Henneman and his colleagues. This same size-dependent branching accounts for the rematching of motoneuron size and muscle unit size in reinnervated muscles. Experiments were carried out to determine whether the daily amount of neuromuscular activation of motor units accounts for the size-dependent organization and reorganization of motor unit properties. The normal size-dependent matching of motoneurons and their muscle units with respect to the numbers of muscle fibers per motoneuron was unaltered by synchronous activation of all of the motor units with the same daily activity. Hence, the restored size relationships and rematching of motoneuron and muscle unit properties after nerve injuries and muscle reinnervation sustain the normal gradation of muscle force during movement by size-ordered recruitment of motor units and the process of rate coding of action potentials. Dynamic modulation of size of muscle fibers and their contractile speed and endurance by neuromuscular activity allows for neuromuscular adaptation in the context of the sustained organization of the neuromuscular system according to the size principle.     

Appearance of "transitional" motor units in overloaded rat skeletal muscle

The contractile characteristics of single motor units, isolated from rat plantaris muscles subjected to short-term (30 days) compensatory overload, were assessed to determine whether motor units in transition could be detected. In the control plantaris 88% of the motor units were classified as fast. After overload, a large decline (26.5%) in the proportion of typical fast motor units was noted. The estimated contribution of fast fatigable units to whole muscle tetanic tension (Po 200) also declined (from 55 to 25%), whereas that of fast intermediate motor units increased (from 33 to 55%). In the overloaded plantaris, motor units that exhibited unusual "sag" and contraction time characteristics were detected. These motor units, which could be further subdivided into two distinct types by a variety of indexes, exhibited characteristics intermediate to fast and slow units and therefore were termed "transitional." Transitional units accounted for 12% of the estimated whole muscle Po200 after overload. These experiments characterize novel classifications of motor units undergoing transformation and further detail the motor unit shift that accompanies compensatory overload.     

ATP concentrations and muscle tension increase linearly with muscle contraction.

Previous studies have suggested that activation of ATP-sensitive P2X receptors in skeletal muscle play a role in mediating the exercise pressor reflex (Li J and Sinoway LI. Am J Physiol Heart Circ Physiol 283: H2636-H2643, 2002). To determine the role ATP plays in this reflex, it is necessary to examine whether muscle interstitial ATP (ATPi) concentrations rise with muscle contraction. Accordingly, in this study, muscle contraction was evoked by electrical stimulation of the L7 and S1 ventral roots of the spinal cord in 12 decerebrate cats. Muscle ATPi was collected from microdialysis probes inserted in the muscle. ATP concentrations were determined by the HPLC method. Electrical stimulation of the ventral roots at 3 and 5 Hz increased mean arterial pressure by 13 +/- 2 and 16 +/- 3 mmHg (P < 0.05), respectively, and it increased ATP concentration in contracting muscle by 150% (P < 0.05) and 200% (P < 0.05), respectively. ATP measured in the opposite control limb did not rise with ventral root stimulation. Section of the L7 and S1 dorsal roots did not affect the ATPi seen with 5-Hz ventral root stimulation. Finally, ventral roots stimulation sufficient to drive motor nerve fibers did not increase ATP in previously paralyzed cats. Thus ATPi is not largely released from sympathetic or motor nerves and does not require an intact afferent reflex pathway. We conclude that ATPi is due to the release of ATP from contracting skeletal muscle cells.     

The dependence of the twitch course of medial gastrocnemius muscle of the rat and its motor units on stretching of the muscle.

The relationship between the force of a single twitch of the medial gastrocnemius muscle of the rat and contraction and half-relaxation times, on one hand, and the load of the muscle on the other, was studied. Twitches of the whole muscle and its individual motor units were induced. The optimal load, at which the majority of motor units reached the greatest twitch force, was 10 G. Mean optimal loads for twitches of different types of motor units were very similar. Slow motor units reached a slightly greater twitch force at greater loads (12.5 G) than at 10 G. However, the optimal load for the twitch of the whole muscle was much greater. It was 47 G on the average. The contraction and half-relaxation times of motor units, as well as of the whole muscle, became longer as the force stretching the muscle increased. Half-relaxation time changed more rapidly than contraction time. Both parameters were undergoing the greatest changes in slow motor units.     

Myosin transducer mutations differentially affect motor function, myofibril structure, and the performance of skeletal and cardiac muscles

Striated muscle myosin is a multidomain ATP-dependent molecular motor. Alterations to various domains affect the chemomechanical properties of the motor, and they are associated with skeletal and cardiac myopathies. The myosin transducer domain is located near the nucleotide-binding site. Here, we helped define the role of the transducer by using an integrative approach to study how Drosophila melanogaster transducer mutations D45 and Mhc(5) affect myosin function and skeletal and cardiac muscle structure and performance. We found D45 (A261T) myosin has depressed ATPase activity and in vitro actin motility, whereas Mhc(5) (G200D) myosin has these properties enhanced. Depressed D45 myosin activity protects against age-associated dysfunction in metabolically demanding skeletal muscles. In contrast, enhanced Mhc(5) myosin function allows normal skeletal myofibril assembly, but it induces degradation of the myofibrillar apparatus, probably as a result of contractile disinhibition. Analysis of beating hearts demonstrates depressed motor function evokes a dilatory response, similar to that seen with vertebrate dilated cardiomyopathy myosin mutations, and it disrupts contractile rhythmicity. Enhanced myosin performance generates a phenotype apparently analogous to that of human restrictive cardiomyopathy, possibly indicating myosin-based origins for the disease. The D45 and Mhc(5) mutations illustrate the transducer's role in influencing the chemomechanical properties of myosin and produce unique pathologies in distinct muscles. Our data suggest Drosophila is a valuable system for identifying and modeling mutations analogous to those associated with specific human muscle disorders.     

Use of motor cortex stimulation to measure simultaneously the changes in dynamic muscle properties and voluntary activation in human muscles.

Force responses to transcranial magnetic stimulation of motor cortex (TMS) during exercise provide information about voluntary activation and contractile properties of the muscle. Here, TMS-generated twitches and muscle relaxation during the TMS-evoked silent period were measured in fresh, heated, and fatigued muscle. Subjects performed isometric contractions of elbow flexors in two studies. Torque and EMG were recorded from elbow flexor and extensor muscles. One study (n = 6) measured muscle contraction times and relaxation rates during brief maximal and submaximal contractions in fresh and fatigued muscle. Another study (n = 7) aimed to 1) assess the reproducibility of muscle contractile properties during brief voluntary contractions in fresh muscle, 2) validate the technique for contractile properties in passively heated muscle, and 3) apply the technique to study contractile properties during sustained maximal voluntary contractions. In both studies, muscle contractile properties during voluntary contractions were compared with the resting twitch evoked by motor nerve stimulation. Measurement of muscle contractile properties during voluntary contractions is reproducible in fresh muscle and reveals faster and slower muscle relaxation rates in heated and fatigued muscle, respectively. The technique is more sensitive to altered muscle state than the traditional motor nerve resting twitch. Use of TMS during sustained maximal contractions reveals slowing of muscle contraction and relaxation with different time courses and a decline in voluntary activation. Voluntary output from the motor cortex becomes insufficient to maintain complete activation of muscle, although slowing of muscle contraction and relaxation indicates that lower motor unit firing rates are required for fusion of force.     

Axonal conduction velocity of motor units of rat's medial gastrocnemius muscle

Axonal conduction velocity and its relations to different contractile properties of motor units of medial gastrocnemius muscle were investigated in nine Wistar rats anaesthetized with pentobarbitone. Functionally isolated motor units were identified as slow (S), fast resistant (FR) and fast fatigable (FF). Axons of S motor units conducted significantly more slowly than of fast units, while there was considerable overlap between conduction velocities measured for FR and FF types. The mean values of conduction velocity were 50.9 m/s for S, 68.9 m/s for FR and 71.3 m/s for FF type motor units. Strong and significant negative correlation between conduction velocity and contraction time as well as half-relaxation time was demonstrated. However, only a weak correlation between conduction velocity and twitch tension, tetanic tension or fatigue index was found. The multiple regression analysis revealed that the major factor to determine conduction velocity was contraction time.