Computer simulation study of the shape of motor unit action potential

Motor unit action potentials (MUAPs) of brachial biceps were simulated. A simulated MUAP was obtained as a sum of single fibre action potentials (SFAPs) from all the muscle fibres of a motor unit (MU). The influence of the following factors on MUAP shape for different kinds of recording electrode was studied: fibre density, neuromuscular jitter, temporal dispersion and electrode displacements. The simulation confirms that typical MUAPs recorded with needle electrodes from muscles of low fibre density such as brachial biceps are usually triphasic. Increased fibre density produces MUAPs of more complex shape and higher amplitude. Normal neuromuscular jitter is responsible for the variability of shape of subsequent potentials from the same MU as well as for electromyographic shimmer. Pathologic (increased) jitter makes the shapes of subsequent potentials unrecognizable. The influence of temporal dispersion is interconnected with other factors but rather of minor importance. The simulation shows how big changes in MUAP shape can be expected due to electrode displacements during single experiment or during estimation of MU territory.     

Differentiation of muscle fibers of various types during postnatal development of the ventral serrate muscle in a rat]

In the denticulate ventral muscle of Wistar rats at the age of 1 day--2 months activity of NAD-N-dehydrogenase, succynic dehydrogenase and cytochrome oxidase has been determined in transversal cryostat sections. Quantitative estimation of the enzymes activity has been carried on by the plag-method. With age, general tendency to increasing activity of the enzymes mentioned is noted, but the dynamics of the increase is peculiar for every enzyme. Analysing the histograms on muscular fibre distribution according to their optical density, it is possible to estimate the dynamics. Simultaneously, the width of variational series, central statistical moments, indices of asymmetry and excess are also estimated. The whole course of the muscular fibre development, in accordance to the range and moments of distribution, can be devided into four main stages: stable, initial stage, slow increase of events, rapid increase of events and stabilization of the process. The stages mentioned pass gradually one into another making it possible to mark transitional stages (5--11, 15--19, 34--60 days). Using standard indices, it is possible to characterize more strictly the processes occurring in the course of muscular fibre differentiation. Lack of parallelism in the dynamics of asimilarity and excess can be treated as variety in differentiation of muscular fibres with middle and large optic density, and parallelism in dynamics--as their simultaneous differentiation. By comparing the curves it can be concluded that up to the 12--14th days, variety in differentiation of muscular fibres occurs, while after the 14th day their differentiation is more regular and simultaneous. The method of cytophotometry with subsequent mathematical processing of the results helps to determine the stages of muscular fibres differentiation.     

A study of the motor unit action potential by means of computer simulation

In order to study the motor unit action potential a computer simulation model was developed. It is based on the superposition of single muscle fibre potentials of the fibres belonging to the motor unit. The parameters which characterize each fibre (spatial position, diameter, and a dispersion of arrival time of the potential at the electrode) are chosen from statistical distributions which can be derived from anatomical and physiological data. The electrode type, position and dimensions can be specified. Simulated motor unit action potentials are presented in the time and frequency domain. The simulation results refer to (1) the influence of the electrode position and dimensions with respect to the motor unit territory, (2) the meaning of this model for the study of pathological phenomena, (3) the variability of some parameters characterizing the motor unit, (4) the selectivity of uni- and bipolar electrodes and finally (5) the influence of the geometrical situation of the motor end-plates within the muscle, on the shape of motor unit action potentials.     

Neuromuscular organization of avian flight muscle: architecture of single muscle fibres in muscle units of the pectoralis (pars thoracicus) of pigeon (Columba livia)

The M. pectoralis (pars thoracicus) of pigeons (Columba livia) is comprised of short muscle fibres that do not extend from muscle origin to insertion but overlap ''in-series''. Individual pectoralis motor units are limited in territory to a portion of muscle length and are comprised of either fast twitch, oxidative and glycolytic fibres (FOG) or fast twitch and glycolytic fibres (FG). FOG fibres make up 88 to 90% of the total muscle population and have a mean diameter one-half of that of the relatively large FG fibres. Here we report on the organization of individual fibres identified in six muscle units depleted of glycogen, three comprised of FOG fibres and three comprised of FG fibres. For each motor unit, fibre counts revealed unequal numbers of depleted fibres in different unit cross-sections. We traced individual fibres in one unit comprised of FOG fibres and a second comprised of FG fibres. Six fibres from a FOG unit (total length 15.45 mm) ranged from 10.11 to 11.82 mm in length and averaged (± s.d.) 10.74 ± 0.79 mm. All originated bluntly (en mass) from a fascicle near the proximal end of the muscle unit and all terminated intramuscularly. Five of these ended in a taper and one ended bluntly. Fibres coursed on average for 70% of the muscle unit length. Six fibres from a FG unit (total length 34.76 mm) ranged from 8.97 to 18.38 mm in length and averaged 15.32 ± 3.75 mm. All originated bluntly and terminated intramuscularly; one of these ended in a taper and five ended bluntly. Fibres coursed on average for 44% of the muscle unit length. Because fibres of individual muscle units do not extend the whole muscle unit territory, the effective cross-sectional area changes along the motor unit length. These non-uniformities in the distribution of fibres within a muscle unit emphasize that the functional interactions within and between motor units are complex.     

Simulation analysis of the ability to estimate motor unit propagation velocity non-invasively by different two-channel methods and types of multi-electrodes.

Ability to estimate motor unit propagation velocity correctly using different two-channel methods for delay estimation and different non-invasive spatial filters was analysed by simulation. It was established that longitudinal double difference electrodes could be not a better choice than simple bipolar parallel electrodes. Spatial filtration with a new multi-electrode (performing difference between signals detected by two transversal double difference electrodes positioned along the muscle fibres) promises to give the best estimate. Delay estimation between reference points is more preferable than that based on the cross-correlation technique, which is considerably sensitive to the fundamental properties of the muscle fibre extracellular fields. Preliminary averaging and approximation of the appropriate parts of the signals around chosen reference points could reduce the larger noise sensitivity and the effects of local tissue inhomogeneities as well as eliminate the sampling problem. A correct estimate of the propagation velocity could be impossible, even in the case of not very deep motor units (15 or 10 mm, depending on the spatial filter used) with relatively long (about 120 mm) muscle fibres. In the case of fibres with asymmetrical location of the end-plates in respect to the fibre ends, the propagation velocity estimates could be additionally biased above the longer semilength of the motor unit fibres.     

Muscle fibre growth dynamics in diploid and triploid rainbow trout

The effect of triploidy on muscle fibre growth was determined by comparing hyperplasia and hypertrophy of white muscle fibres in all-female, diploid and triploid rainbow trout Oncorhynchus mykiss (100–400 mm total length). Conventional morphometry and protein and DNA concentrations were used to assess muscle fibre hyperplasia and hypertrophy in white muscle samples derived from an anterio-dorsal location. Muscle fibre distributions were significantly different between triploids and diploids in trout <300 mm. The proportion of fibres <20 μm was higher in diploids than in triploids and the proportion of fibres in the 20–40 μm category was higher in triploids than in diploids. This indicates that the hyperplastic fibres of triploids are larger than those of diploids. Larger hyperplastic fibres in triploids are probably due to the combined effect of increased nuclear size in triploids and the relatively high nucleus: cell ratio observed in small muscle fibres. These larger fibres may be less favourable to cellular metabolic exchange because of their smaller surface area to volume ratios, and perhaps account for reduced viability and growth observed in triploids during early life stages. On the other hand, the lack of difference in the distribution of fibres <20 μm between diploids and triploids at larger body size ranges (301–400 mm) imply that triploid trout may have higher rates of new fibre recruitment and growth capacity at these sizes. There was no difference between diploid and triploid trout in the mean size of muscle fibres; however, the number of fibres per unit area was reduced by 10% in triploids. No differences were observed in protein or DNA concentrations in muscle tissues between the two genetic groups. Since triploid nuclei have 1·5 times more DNA than diploid nuclei, this deviation from the expected muscle DNA concentration (1·3–1·4 times more DNA in triploids when the 10% reduction in fibre density is considered) suggests that the number of nuclei per muscle fibre is reduced. In both diploids and triploids, mean fibre size increased with body length while fibre density decreased. Similarly, protein concentration in the muscle tissue increased and DNA concentration declined with increasing body length. Protein/DNA ratio was strongly and positively correlated with fibre size. These results demonstrate that changes in DNA and protein concentrations can be used to assess hyperplasia and hypertrophy in muscle tissues. However, the morphometric procedure provides better insight into muscle fibre growth as it enables the direct visualization and analysis of muscle fibre distribution patterns.     

Spectral and time domain characteristics of single muscle fibre action potentials during continuous activity extracted from model considerations

A model of the muscle fibre extracellular action potentials (ECAPs) calculation using experimentally recorded intracellular action potentials (ICAPs) has been applied to investigate the effect of repetitive stimulation on the electrical activity of isolated frog muscle fibres. The ECAPs were calculated both at small (0.01 mm) and at large (5 mm) radial distances to the fibre axis, and their relationship with the original ICAP parameters has been inferred. Fourier transformation of the calculated ECAPs in order to obtain the spectral characteristics and to trace out their behaviour during continuous fibre activity was performed. Stimulation frequency dependence on the ECAP time characteristics and on the shift of the maximum spectral density towards low frequencies at small and large radial distance were observed. The spectral density peak frequency is propagation velocity (PV)-dependent. The advantage of the presented method over the available experimental extracellular recording techniques from isolated muscle fibers is the possibility to show the effect of continuous muscle fibre activity on the parameters of the ECAPs and their spectral characteristics at large radial distance, which is not experimentally accessible. Our results are in agreement with those experimentally obtained. The results from the model prove the role of changes in PV of excitation along the muscle fibres (representing the last link in the complex organized motor system) in the development of fatigue. Received: 24 July 1997 / Accepted in revised form: 2 July 1998     

Bootstrap resampling for voxel-wise variance analysis of three-dimensional density maps derived by image analysis of two-dimensional crystals

Difference density maps are commonly used in structural biology for identifying conformational changes in macromolecular complexes. For interpretation of the results, it is essential to estimate the variance or standard deviation of the difference density and the distribution of errors in space. In order to compare three-dimensional density maps of gap junction channels with and without the C-terminal regulatory domain, we developed a bootstrap resampling method for estimation of the voxel-wise standard deviation. The bootstrap approach has been successfully used for estimating the sampling distribution from a limited data set and for estimating the statistical properties of the derived quantities [Efron, B., 1979. Bootstrap methods: another look at the jackknife. Ann. Stat. 7, 1-26]. In our application, the standard deviation map can be estimated by bootstrapping the images. Our results show that, apart from the symmetry axes and small regions bordering the lumen of the extracellular vestibule, difference maps normalized by the mean of the standard deviation map can be used as a good approximation of the t-test map of the gap junction crystals.     

The overlapping in the striatum of the projection areas of corticofugal fibers from the somatosensory and motor regions of the rat cerebral cortex]

The Fink-Heimer techniques were used to determine the neostriatal projections from cortical M1 and S1 physiologically identified representations of the forepaw. While corticostriatal fibres from S1 enter dorso-lateral parts of the neostriatum fibres from M1 penetrate the dorsal part of the neostriatum more rostrally. Both fibre groups pass in rostrocaudal direction to the globus pallidum, forming terminals on their way. Most of the fibre terminals of M1 projection are located laterally overlapping the zone of distribution of S1 fibre terminals in the neostriatum. Overlapping terminals of the corticofugal fibres of S1 and M1 in the lateral neostriatum can form a basis for interaction of somatosensory and motor impulses. It may be suggested that the role of the neostriatum in sensory integration is associated with certain features of the corticofugal fibre terminals distribution in it at all levels of development of the central nervous system.     

Statistical Analysis of Motor Unit Firing Patterns in a Human Skeletal Muscle

A statistical analysis of the firing pattern of single motor units in the human brachial biceps muscle is presented. Single motor unit spike trains are recorded and analyzed. The statistical treatment of these spike trains is as stochastic point processes, the theory of which is briefly discussed. Evidence is presented that motor unit spike trains may be modelled by a renewal process with an underlying gaussian probability density. Statistical independence of successive interspike intervals is shown using scatter diagrams; the hypothesis of a gaussian distribution is accepted at the 99th percentile confidence limit, chi-square test, in 90% of the units tested. A functional relationship between the mean and standard deviation is shown and discussed; its implications in obtaining sample size are presented in an appendix.The results of higher order analysis in the form of autocorrelograms and grouped interval histograms are presented. Grouped interval histograms are discussed in the context of motor unit data, and used to confirm the hypothesis that a stable probability density function does not represent a good model of the data at this level of analysis.     

The staining of elastic fibres with Direct Blue 152. A general hypothesis for the staining of elastic fibres

Summary Elastic fibres may be stained by a number of dyes, e.g. Direct Blue 1 (C.I. 24410), Direct Blue 10 (C.I. 24340), Direct Blue 15 (C.I. 24400), Direct Blue 152 (C.I. 24366) and Direct Violet 37 (C.I. 24370). A convenient method using Direct Blue 152 has been developed which is specific for elastic fibres. The method is simple and allows the demonstration of other connective tissue fibres. Staining of elastic fibres is unimpaired by numerous blocking procedures or by changes in dyebath pH. These properties are shared by several standard elastic fibre stains.As the Direct dyes and several of the standard elastic fibre stains possess numerous aromatic rings a wide range of dyes containing varying numbers of aromatic rings were examined for ability to stain elastic fibres. No association was observed between the ability to stain elastic fibres and dye class, formal charge or the presence of hydrogen bonding groups. Staining was, however, definitely associated with the presence in the dye molecule of 5 or more aromatic rings. This suggested that van der Waals forces of attraction may be responsible for elastic fibre staining both by Direct dyes and the standard elastic fibre stains. Staining of elastic fibres as a side-effect in many procedures is similarly explicable.     

Intrafibre distribution of succinate dehydrogenase in cat tibialis anterior motor units.

Using isolated ventral root filament stimulation and glycogen depletion techniques, 14 motor units from the cat tibialis anterior were studied. Based on their mechanical properties, the units were classified as either slow-fatigue resistant, fast-fatigue resistant, fast-fatigue intermediate, or fast-fatigable. Quantitative histochemical and computer assisted image analysis techniques were used to determine the activity of succinate dehydrogenase in a population of fibres in each unit. In addition, the intrafibre distribution of succinate dehydrogenase activity was measured in those same fibres by calculating the enzymatic activity of circumferential layers every 0.5 microns starting from the fibre edge to its centre. It was established that enzymatic activity and radial distance were linearly related in the fibres. A range in succinate dehydrogenase activity (mean coefficient of variation, 29%) was observed among the fibres of a unit. In contrast, the intrafibre distribution of that activity was rather consistent (mean variation, 4%) across the fibres of a unit. Further, the intrafibre distribution was similar among the fibres of units classified as the same type. However, the intrafibre distribution was disparate among the different unit types. These data suggest that the intrafibre distribution of mitochondrial enzymes may contribute to the mechanical properties of a motor unit. In this regard, a hypothesis is proposed that describes how the absolute activity of a mitochondrial enzyme, and the intrafibre distribution of that activity, may interactively contribute to the fatigue resistance of a unit.     

Selective deafferentation of hand cutaneous territory is followed by changes in fibre type distribution of a forearm muscle in the horse

Based on previous observations that capsaicin can selectively damage group III and IV afferents and induce muscle fibre transformation, we hypothesized that eliminating, by means of capsaicin, the group III and IV afferents of a peripheral territory it could lead to a fibre transformation in a muscle involved in the flexor reflexes of the same peripheral territory. Therefore, capsaicin was injected into the palmar nerves of the forelimb of the horse to investigate if eliminating group III and IV afferents from the hand of the horse a muscle fibre transition would occur in the flexor carpi radialis (FCR) muscle, which is involved in the flexor reflexes of the finger itself. 120 days after capsaicin injection, type I slow fibres increased and type IIA fast fibres decreased. We presume that the long lasting deafferentation of the ergo-nociceptive fibres causes a plastic remodelling in the central nervous system and indirectly influences the motoneuron excitability via short or long loop-pathways enhancing their tonic discharge.     

A distributed-parameter model of the myelinated human motor nerve fibre: temporal and spatial distributions of action potentials and ionic currents

 A double cable model of the myelinated human motor nerve fibre is presented. The model is based on the nodal and internodal channels in a previous, two-component model of human motor axons (Bostock et al. 1991), added to a complex extended cable structure of nodal, paranodal and internodal segments. The model assumes a high-resistance myelin sheath and a leakage pathway to the internodal axolemma via the paranodal seal resistance and periaxonal space. The parameter values of the model were adjusted to match the recordings of threshold electrotonus in human motor fibres from Bostock et al. (1991). Kirchoff ’s current law was used to derive a system of partial differential equations for the electrical equivalent circuit, and numerical integration was performed with a fixed time increment and non-uniform spatial step sizes, in accordance with the complex structure of the fibre. The model calculations provide estimates of the spatial and temporal distributions of action potentials and their transaxonal and transmyelin components, both in different segments of the fibre and at different moments during action potential propagation. The distribution of transaxonal and transmyelin currents along the fibre and their contributions from different ionic channels are also explored. Received: 14 July 1994/Accepted in revised form: 4 April 1995     

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.     

Motor unit action potential field — Modelling results

The theoretical field of a motor unit (MU) action potential (MUP) was considered. It has been proved that in normal muscles the shape of a smooth threephasic MUP curve is determined mostly by the spatial distribution of MU muscle fibres. Phenomena called time dispersion are of prime importance in pathology, where they complicate normal threephasic MUP.Amplitudes and time parameters of model three-phasic MUP were analyzed as a function of the radial distance R from the geometrical centre of the motor unit territory (MUT) and approximated by mathematical expressions. It appeared that analysis of radial variability of MUP curve allows conclusions to be made about the MUT size and the spatial distribution of MU muscle fibres. These anatomical features of a MU are often changed in pathological muscles, thus the proposed methods of their evaluation could be helpful in diagnosis of neuromuscular diseases.     

Study of water sorption on modified Agave fibres

Polymer composite materials are usually reinforced by synthetic matter such as carbon or glass fibres. However, owing to their good mechanical properties and low density, natural fibres are now increasingly being considered as reinforcement. With the aim of a new natural fibre based composite, various chemical treatments have been performed on Agave (Americana L.) fibres in order to improve their compatibility with the polymer matrix and to reduce their affinity for water. The effect of these treatments on the fibre water sorption power has been investigated by means of a micro-balance. Equilibrium water sorption isotherms have been deduced from weight variations of the fibres under water vapor pressure increments. Several specific physico-chemical models have been tested to describe the water sorption isotherms. The Park’s model was found to describe the experimental results accurately and over a wide activity range. The sorption kinetics was also exploited in order to evaluate the diffusivity of water in the fibres. The variation of the water diffusion coefficient with water concentration is in agreement with the triple sorption mode described by the Park’s model. These results show a global increase of moisture resistance of the fibres after chemical treatment. This effect is interpreted in terms of chemical and structural modifications of the cell-wall structure.     

The effect of activity during early postnatal development on motor unit size

At early stages of neuromuscular development, motor unit territory is expanded, with each muscle fibre being supplied by several axons. During postnatal development, some synapses are eliminated, motor unit size decreases, and the adult distribution of motor unit sizes emerges. This process depends on activity, since it proceeds more rapidly when the nerve is activated and is slower when activity is reduced. Here we studied whether, in addition to influencing the rate of retraction of motor unit territory, activity during the critical period of development affects the final outcome of the distribution of motor unit sizes. The sciatic nerve of 8- to 12-day-old rats was stimulated daily. One week later the tension of the extensor digitorum longus muscle and that of its individual motor units was recorded. The sizes of individual motor units were calculated and compared with those from animals that received no stimulation. The distribution of motor unit sizes from stimulated muscles was not significantly different from those from control muscles. Therefore, we conclude that although activity increases the rate at which motor units attain their adult size, it does not influence the final outcome of motor unit size distribution.     

Intrinsic innervation of the rat knee joint articular capsule and ligaments

In spite of the practical importance of having a detailed knowledge of knee joint innervation to understand the pathophysiologic aspects, little information is now available concerning the density and pattern of the nerve fibres which are distributed to it. The present study has been designed to investigate the density and distribution of nerve fibres and receptor corpuscles in the knee joint articular capsule, cruciate and collateral ligaments in the rat, using the acetylcholinesterase (AChE) histochemical in toto staining technique. The investigation was performed on male Wistar rats of 3 months of age, some of which had been treated with capsaicin to deplete their afferent 'C' fibres of their content of neuropeptides. AChE-positive nerve fibres and different types of receptor corpuscle endings were found within articular capsule and ligaments. The highest density of AChE-positive nerve fibres was noticeable in the fibular collateral ligament followed by the tibial collateral ligament, the posterior cruciate ligament, the anterior cruciate ligament and the articular capsule. In the articular capsule the number of type I endings was higher than in the ligaments. The opposite is true for the other type of receptor corpuscles found as well as for nerve endings. Capsaicin treatment significantly reduced the density of AChE-positive nerve fibres in knee joint ligaments but did not affect nerve fibres in the articular capsule. Moreover, it caused the disappearance of some kind of receptor corpuscles within the collateral and cruciate ligaments. The above data collectively suggest that the AChE in toto staining technique may represent a good method for investigating joint innervation and that a significant percentage of nerve fibres supplying knee joint ligaments is represented by C fibre afferents.