Distribution and length of muscle spindles and their 3D visualisation in the medial gastrocnemius of male and female rats.

The spatial distribution of the medial gastrocnemius muscle spindles of 10 male and 10 female rats was analysed under a light microscope, and for the first time, visualised using a 3D model of the muscle. Serial cross-sections of the medial gastrocnemius muscles were separated into 10 divisions along with the proximo-distal axis. The muscle spindles of the rat medial gastrocnemius were predominantly distributed on the proximo-medial divisions of the muscle. There were no sex-related differences in the distribution of the studied receptors. A single division contained an average of 2.71 receptors for animals of both sexes. Moreover, the calculated lengths of male and female rat muscle spindles were comparable, and average lengths did not significantly differ (3.30 ± 1.47 mm for male and 3.26 ± 1.32 mm for female rats). Therefore, the present results fill gaps in recent observations concerning similarities in muscle spindle numbers between male and female animals, despite significant differences in muscle mass and size.

The contractile properties of motor units in the rat flexor digitorum brevis muscle have continuous distribution.

The majority of motor unit studies were performed predominantly on calf muscles, where three types of units: S, FR and FF were found. These muscles are involved in postural activity, walking, running and jumping. The properties of foot muscles that perform other functions, e.g. scratching (in animals), and are purely co-active with calf muscles, are poorly known. The aim of the present study was to investigate the contractile properties of motor units in the flexor digitorum brevis. Fifty-six motor units were studied in male Wistar rats. Several methods of fast/slow motor unit categorization, presence of sag, contraction time values, and 20 Hz index, did not allow the separation of the studied motor units into discrete clusters. Therefore, motor units were divided into two groups: fatigable and resistant to fatigue, based on the fatigue index with the border value of 0.5 (although the distribution of the index was not bimodal). The fatigable motor units were stronger and faster compared to the resistant ones. In conclusion, the distribution of motor unit contractile properties in the studied foot muscle was continuous and indicated a lack of three separate physiological types of motor units that usually occurs for the majority of hindlimb muscles. This discrepancy appears to be associated with differences in the typical forms of motor unit activity in distinct muscles.

Factors contributing to sag in unfused tetanic contractions of fast motor units in rat medial gastrocnemius.

The sag phenomenon can be observed in fast motor units (MUs) as a transitional decline in force during unfused tetanic contractions; however, its mechanisms are poorly understood. The study aimed to identify in the rat muscle factors that contribute to sag in two types of fast MUs: fast fatigable (FF) and fast resistant to fatigue (FR). First, we performed mathematical decomposition of sagging tetanic contractions of FF and FR MUs into twitch-like responses to consecutive stimuli. This process indicated an increase in the amplitudes of a few initial responses (up to the 2nd-3rd for FF and up to the 2nd-7th for FR MUs), followed by a decrease in the amplitudes of later responses. In comparison to the first twitch, the relative increase in force amplitudes of the several subsequent decomposed responses was smaller, and their contraction and relaxation times were shorter for FF than for FR units, which corresponded to observed differences in their sag profiles. Additionally, after occlusion of the blood circulation, sag disappeared, but it reappeared after restoration of the blood supply. This indicates that the presence of sag depends on the proper circulation in the muscle.

A model of the rat medial gastrocnemius muscle based on inputs to motoneurons and on an algorithm for prediction of the motor unit force.

The muscle force is the sum of forces of multiple motor units (MUs), which have different contractile properties. During movements, MUs develop unfused tetani, which result from summation of twitch-shape responses to individual stimuli, which are variable in amplitude and duration. The aim of the study was to develop a realistic muscle model that would integrate previously developed models of MU contractions and an algorithm for the prediction of tetanic forces. The proposed model of rat medial gastrocnemius muscle is based on physiological data: excitability and firing frequencies of motoneurons, contractile properties, and the number and proportion of MUs in the muscle. The MU twitches were modeled by a six-parameter analytical function. The excitability of motoneurons was modeled according to a distribution of their rheobase currents measured experimentally. Processes of muscle force regulation were modeled according to a common drive hypothesis. The excitation signal to motoneurons was modeled by two form types: triangular and trapezoid. The discharge frequencies of MUs, calculated individually for each MU, corresponded to those recorded for rhythmic firing of motoneurons. The force of the muscle was calculated as the sum of all recruited MUs. Participation of the three types of MUs in the developed muscle force was presented at different levels of the excitation signal to motoneurons. The model appears highly realistic and open for input data from various skeletal muscles with different compositions of MU types. The results were compared with three other models with different distribution of the input parameters. NEW & NOTEWORTHY The proposed mathematical model of rat medial gastrocnemius muscle is highly realistic because it is based strictly on experimentally determined motor unit contractile parameters and motoneuron properties. It contains the actual number and proportion of motor units and takes into consideration their different contributions to the whole muscle force, depending on the level of the excitation signal. The model is open for input data from other muscles, and additional physiological parameters can also be included.

Sense of extension force and angle of the knee joint are correlated between two generations of men.

Numerous motor abilities depend on the activity of proprioceptors, which has been suggested to be genetically determined. To test this hypothesis, the control of torque generated by knee extensors and knee position was studied in 30 father-son pairs both before and immediately after running. After stabilisation of the participant in a sitting position, the knee joint of his dominant leg was flexed to 90°, and the maximal voluntary torque (MVT) of the dominant knee extensors under static conditions was measured. The participant then tried five times to produce 50% of the MVT. Next, the participant extended the knee to 45° five times without visual control. Significant correlations between the reproducibility of successive trials for groups of fathers and their sons were found. The correlation coefficients for the repeatability of the knee extension torque were 0.69 (confidence interval [CI] = 0.45-0.84; P < 0.01) and 0.75 (CI = 0.54-0.87; P < 0.01) before and after the fatiguing exercise, respectively, whereas the coefficient for the reproducibility of positioning the knee was 0.49 (CI = 0.16-0.72; P < 0.01) after the fatiguing exercise. Our results indicate a significant influence of hereditary factors on the control of limb torque and position.

Changes in contractile properties and action potentials of motor units in the rat medial gastrocnemius muscle during maturation.

The early phase of development of muscles stops following the disappearance of embryonic and neonatal myosin and the elimination of polyneuronal innervation of muscle fibres with the formation of motor units (MUs), but later the muscle mass still considerably increases. It is unknown whether the three types are visible among newly formed MUs soon after the early postnatal period and whether their proportion is similar to that in adult muscle. Moreover, the processes responsible for MU-force regulation by changes in motoneuronal firing rate as well as properties of motor unit action potentials (MUAPs) during maturation are unknown. Three groups of Wistar rats were investigated - 1 month old, 2 months old and the adult, 9 months old. The basic contractile properties and action potentials of MUs in the medial gastrocnemius (MG) muscle were analysed. The three types of MUs were distinguishable in all age groups, but higher proportion of slow MUs was noticed in young rats (29%, 18% and 11% in 1, 2 and 9 months rats, respectively). The fatigue index for fast fatigable MUs in 1 month old rats was about 2 times higher than in 9 months old rats. The twitch time parameters of fast MUs were shortened during the maturation; for these units, the force-frequency curves in young rats were shifted towards lower frequencies, which suggested that fast motoneurons of young animals generate lower firing rates. Higher twitch-to-tetanus ratios noted for the three MU types in young rats suggested the smaller role of rate coding in force regulation processes, and the higher role of MU recruitment in young rats. No significant differences in MUAP parameters between two groups of young and adult animals were observed. Concluding, the maturation process evokes deeper changes in fast MUs than in slow ones.

Adaptations of motoneuron properties to chronic compensatory muscle overload.

The aim of the study was to determine whether chronic muscle overload has measurable effect on electrophysiological properties of motoneurons (MNs), and whether duration of this overload influences intensity of adaptations. The compensatory overload was induced in the rat medial gastrocnemius (MG) by bilateral tenotomy of its synergists (lateral gastrocnemius, soleus, and plantaris); as a result, only the MG was able to evoke the foot plantar flexion. To assure regular activation of the MG muscle, rats were placed in wheel-equipped cages and subjected to a low-level treadmill exercise. The intracellular recordings from MG motoneurons were made after 5 or 12 wk of the overload, and in a control group of intact rats. Some of the passive and threshold membrane properties as well as rhythmic firing properties were considerably modified in fast-type MNs, while remaining unaltered in slow-type MNs. The significant changes included a shortening of the spike duration and the spike rise time, an increase of the afterhyperpolarization amplitude, an increase of the input resistance, a decrease of the rheobase, and a decrease of the minimum current necessary to evoke steady-state firing. The data suggest higher excitability of fast-type MNs innervating the overloaded muscle, and a shift towards electrophysiological properties of slow-type MNs. All of the adaptations could be observed after 5 wk of the compensatory overload with no further changes occurring after 12 wk. This indicates that the response to an increased level of chronic activation of MNs is relatively quick and stable.

Sex differences in the number and size of motoneurons innervating rat medial gastrocnemius muscle.

The sex differences in the number and morphometric parameters of motoneurons in motor nuclei are poorly known. The aim of this study was to determine the differences in the number and size of alpha and gamma motoneurons of the medial gastrocnemius (MG) muscle in male and female Wistar rats. Retrogradely labelled cell bodies of motoneurons of 6 months old animals were studied following a bath of the proximal stump of the transected MG nerve in a horseradish peroxidase solution. The number and soma diameters of male and female MG motoneurons were determined from serial microscopic images of sections. The weight of the brain and spinal cord was on average 17% higher in males than in females. The mean number of motoneurons was 13% higher in males than in females and amounted to 94 and 83 motoneurons, respectively. In each case, the average soma diameters and cross-section areas of motoneurons in motor nucleus were distributed bimodally: motoneurons smaller than 27.5 µm in diameter were recognized as gamma and greater ones as alpha motoneurons. In males, the motor nucleus contained on the average 66 alpha motoneurons, whereas in females, 56 alpha motoneurons, that is the mean number of alpha motoneurons was 17% higher in males. Moreover, the soma diameters of gamma and alpha motoneurons were significantly bigger in males and the difference amounted 9 and 6%, respectively. It is concluded that the number as well as size of alpha and size of gamma motoneurons in the MG motor nucleus are greater in males.

Concomitant changes in afterhyperpolarization and twitch following repetitive stimulation of fast motoneurones and motor units.

The study aimed at determining changes in a course of motoneuronal afterhyperpolarization (AHP) and in contractile twitches of motor units (MUs) during activity evoked by increasing number of stimuli (from 1 to 5), at short interspike intervals (5 ms). The stimulation was applied antidromically to spinal motoneurones or to isolated axons of MUs of the medial gastrocnemius muscle within two separate series of experiments on anesthetized rats. Alterations in the amplitude and time parameters of the AHP of successive spikes were compared to changes in force and time course of successive twitches obtained by mathematical subtraction of tetanic contractions evoked by one to five stimuli. The extent of changes of the studied parameters depended on a number of applied stimuli. The maximal modulation of the AHP and twitch parameters (a prolongation and an increase in the AHP and twitch amplitudes) was typically observed after the second pulse, while higher number of pulses at the same frequency did not induce so prominent changes. One may conclude that changes observed in parameters of action potentials of motoneurons are concomitant to changes in contractile properties of MU twitches. This suggests that both modulations of the AHP and twitch parameters reflect mechanisms leading to force development at the beginning of MU activity.

Long-term effects of whole-body vibration on motor unit contractile function and myosin heavy chain composition in the rat medial gastrocnemius.

Structural and physiological mechanisms underling functional adaptations of a muscle to chronic whole-body vibration (WBV) are poorly understood. The study aimed at examining the contractile properties of motor units and the myosin heavy chain (MHC) expression in rat medial gastrocnemius muscle in response to 3- or 6-month periods of the WBV. The three-month WBV induced modifications of contractile properties principally in slow (S) and fast resistant to fatigue (FR) motor units. In S units an increase in the maximum tetanus force, a reduction in the twitch force and a decrease in the twitch-to-tetanus force ratio were found. In FR units a shortening in the twitch time parameters, a decrease in the twitch-to-tetanus ratio and an increase in the fatigue resistance were observed. In addition, a decrease in the type I and an increase in the type IIax MHC content were revealed. The six-month WBV caused a decrease in the twitch-to-tetanus force ratio in S and FR units. Other structural and physiological changes in MU properties previously seen were no longer apparent. In conclusion, responses to the long-term WBV stimulus vary between particular types of motor units, what suggests that multiple adaptive processes in muscle tissue are involved.

Extra-torque of human tibialis anterior during electrical stimulation with linearly varying frequency and amplitude trains.

This work aimed to characterise the whole human muscle input/output law during electrical stimulation with triangular varying frequency and amplitude trains through combined analysis of torque, mechanomyogram (MMG) and electromyogram (EMG). The tibialis anterior (TA) of ten subjects (age 23-35 years) was investigated during static contraction obtained through neuromuscular electrical stimulation. After potentiation, TA underwent two 15s stimulation patterns: (a) frequency triangle (FT): 2 > 35 > 2 Hz at Vmax (amplitude providing full motor unit recruitment); (b) amplitude triangle (AT): Vmin > Vmax > Vmin (Vmin providing TA least mechanical response) at 35 Hz. 2 > 35 Hz or Vmin > Vmax as well as 35 > 2 Hz or Vmax > Vmin were defined as up-going ramp (UGR) and down-going ramp (DGR), respectively. TA torque, MMG and EMG were detected by a load cell, an optical laser distance sensor and a probe with two silver bar electrodes, respectively. For both FT and AT, only the two mechanical signals resulted always larger in DGR than in UGR, during AT extra-torque and extra-MMG were present even in the first 1/3 of the amplitude range where EMG data presented no significant differences between DGR and UGR. Our data suggest that extra-torque and extra-displacement are evident for both FT and AT, being mainly attributed to an intrinsic muscle property.

The influence of a 5-wk whole body vibration on electrophysiological properties of rat hindlimb spinal motoneurons.

The study aimed at determining the influence of a whole body vibration (WBV) on electrophysiological properties of spinal motoneurons. The WBV training was performed on adult male Wistar rats, 5 days a week, for 5 wk, and each daily session consisted of four 30-s runs of vibration at 50 Hz. Motoneuron properties were investigated intracellularly during experiments on deeply anesthetized animals. The experimental group subjected to the WBV consisted of seven rats, and the control group of nine rats. The WBV treatment induced no significant changes in the passive membrane properties of motoneurons. However, the WBV-evoked adaptations in excitability and firing properties were observed, and they were limited to fast-type motoneurons. A significant decrease in rheobase current and a decrease in the minimum and the maximum currents required to evoke steady-state firing in motoneurons were revealed. These changes resulted in a leftward shift of the frequency-current relationship, combined with an increase in slope of this curve. The functional relevance of the described adaptive changes is the ability of fast motoneurons of rats subjected to the WBV to produce series of action potentials at higher frequencies in a response to the same intensity of activation. Previous studies proved that WBV induces changes in the contractile parameters predominantly of fast motor units (MUs). The data obtained in our experiment shed a new light to possible explanation of these results, suggesting that neuronal factors also play a substantial role in MU adaptation.

Gender differences in morphometric properties of muscle fibres measured on cross-sections of rat hindlimb muscles.

The study was aimed at demonstrating gender differences in the numbers, diameters and cross-section areas of muscle fibres for three hindlimb skeletal muscles responsible for locomotion and maintenance of body posture: soleus, tibialis anterior and flexor digitorum brevis in rats. The experiments were performed on five 6-month-old male and female Wistar rats. In both genders, all studied muscles of the right and left hindlimbs were isolated from surrounding tissues and excised for further procedures. The muscle transverse cross-sections taken from the muscle mid-belly were analysed. Following staining of reticular fibres by silver impregnation, the numbers, diameters and cross-section areas of muscle fibres were determined from microscopic images of muscle sections. The body mass of male rats was 80% higher than that of females. In addition, the muscle mass and the cross-section area were 53-82 and 26-45% higher in males, respectively. The number of muscle fibres was 11-42% higher in males than in females whereas the fibre diameters were 7-29% higher in males. The most conspicuous differences between males and females were found with respect to tibialis anterior, whereas the smallest differences were evident in soleus. The present study revealed that the gender morphometric differences in the studied rat hindlimb muscles were mainly owing to differences in number and size of muscle fibres and that the difference in muscle mass could be explained mainly from higher number of muscle fibres in males and to smaller degree from their larger diameters.

The tetanic depression in fast motor units of mammalian skeletal muscle can be evoked by lengthening of one initial interpulse interval.

A lower than expected tetanic force (the tetanic depression) is regularly observed in fast motor units (MUs) when a higher stimulation frequency immediately follows a lower one. The aim of the present study was to determine whether prolongation of only the first interpulse interval (IPI) resulted in tetanic depression. The experiments were carried out on fast MUs of the medial gastrocnemius muscle in cats and rats. The tetanic depression was measured in each case as the force decrease of a tetanus with one IPI prolonged in relation to the tetanic force at the respective constant stimulation frequency. Force depression was observed in all cases studied and was considerably greater in cats. For cats, the mean values of force depression amounted to 28.64% for FR and 10.86% for FF MUs whereas for rats 9.30 and 7.21% for FR and FF motor units, respectively. Since the phenomenon of tetanic depression in mammalian muscle is commonly observed even after a change in only the initial interpulse interval within a stimulation pattern, it can effectively influence processes of force regulation during voluntary activity of a muscle, when motoneurones progressively increase the firing rate.

Gender differences in the morphometric properties of muscle fibres and the innervation ratio of motor units in rat medial gastrocnemius muscle.

UNLABELLED: With 2 figures and 2 tables SUMMARY: Previous studies of motor unit contractile properties in the rat medial gastrocnemius revealed that these units generate higher forces in males than in females. Therefore, in the present study the number and morphometric parameters of muscle fibres and the innervation ratio of motor units in the medial gastrocnemius muscle were studied in male and female Wistar rats. The study additionally aimed at determining reasons of gender differences in motor unit force parameters, i.e. the number and diameter of muscle fibres, and mean values of the motor unit innervation ratios. Following staining of reticuline fibres by silver impregnation, the number, diameter and cross-section area of muscle fibres were determined on microscopic images of transverse muscle sections. In males, the muscles were approximately 1.5 times larger by mass and contained about 11 800 muscle fibres, whereas in females the muscles contained around 8000 fibres. In addition, the mean diameter and mean cross-section area of muscle fibres were 14 and 29% larger in males, respectively. Based on previously determined numbers of motoneurons innervating the medial gastrocnemius muscle in male and female rats, the mean innervation ratio, i.e. the number of muscle fibres innervated by one motoneuron, was estimated. This ratio was approximately 26% greater in males compared to females, with values of 207 and 153 fibres per motoneuron, respectively. Therefore, the differences in muscle fibre morphometric parameters and in the innervation ratio are responsible for higher forces of motor units in male muscles.

Doublet of action potentials evoked by intracellular injection of rectangular depolarization current into rat motoneurones.

A doublet of action potentials is frequently observed at the beginning of motoneuronal discharge patterns and its appearance leads to a considerable increase in the motor unit force. The aims of this study were (1) to determine the relationship between the intensity of rectangular depolarization currents injected into motoneurones and their ability to generate doublets and (2) to evaluate the influence of the initial doublets on changes in motoneuronal firing frequency. Experiments were performed on anesthetized rats, and recordings were taken from motoneurones located in the L4-L5 segments of the spinal cord. The depolarization current necessary to evoke the initial doublet of action potentials was measured and expressed in multiples of the rheobase. A gradual increase in the intensity of current injected into motoneurones resulted in initial doublets in 80% of the cases studied, at doublet threshold ranges between 1.25 and 4.0 times the rheobase. This suggests that doublets are an effect of strong synaptic excitation of motoneurones that may precede a sudden change in force during a movement. Moreover, in the great majority of the studied motoneurones, this initial doublet caused changes in the subsequent firing rate by the prolongation of the first interspike interval.

Experimentally verified mathematical approach for the prediction of force developed by motor units at variable frequency stimulation patterns.

During normal daily activity, muscle motor units (MUs) develop unfused tetanic contractions evoked by trains of motoneuronal firings at variable interpulse intervals (IPIs). The mechanical responses of a MU to successive impulses are not identical. The aim of this study was to develop a mathematical approach for the prediction of each response within the tetanus as well as the tetanic force itself. Experimental unfused tetani of fast and slow rat MUs, evoked by trains of stimuli at variable IPIs, were decomposed into series of twitch-shaped responses to successive stimuli using a previously described algorithm. The relationships between the parameters of the modeled twitches and the tetanic force level at which the next response begins were examined and regression equations were derived. Using these equations, profiles of force for the same and different stimulation patterns were mathematically predicted by summating modeled twitches. For comparison, force predictions were made by the summation of twitches equal to the first one. The recorded and the predicted tetanic forces were compared. The results revealed that it is possible to predict tetanic force with high accuracy by using regression equations. The force predicted in this way was much closer to the experimental record than the force obtained by the summation of equal twitches, especially for slow MUs. These findings are likely to have an impact on the development of realistic muscle models composed of MUs, and will assist our understanding of the significance of the neuronal code in motor control and the role of biophysical processes during MU contractions.

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, 40Hz unfused tetani, and 150Hz 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.

The image of motor units architecture in the mechanomyographic signal during the single motor unit contraction: in vivo and simulation study.

The mechanomyographic (MMG) signal analysis has been performed during single motor unit (MU) contractions of the rat medial gastrocnemius muscle. The MMG has been recorded as a muscle surface displacement by using a laser distance sensor. The profiles of the MMG signal let to categorize these signals for particular MUs into three classes. Class MMG-P (positive) comprises MUs with the MMG signal similar to the force signal profile, where the distance between the muscle surface and the laser sensor increases with the force increase. The class MMG-N (negative) has also the MMG profile similar to the force profile, however the MMG is inverted in comparison to the force signal and the distance measured by using laser sensor decreases with the force increase. The third class MMG-M (mixed) characterize the MMG which initially increases with the force increases and when the force exceeds some level it starts to decrease towards the negative values. The semi-pennate muscle model has been proposed, enabling estimation of the MMG generated by a single MU depending on its localization. The analysis have shown that in the semi-pennate muscle the localization of the MU and the relative position of the laser distance sensor determine the MMG profile and amplitude. Thus, proposed classification of the MMG recordings is not related to the physiological types of MUs, but only to the MU localization and mentioned sensor position. When the distance sensor is located over the middle of the muscle belly, a part of the muscle fibers have endings near the location of the sensor beam. For the MU MMG of class MMG-N the deflection of the muscle surface proximal to the sensor mainly influences the MMG recording, whereas for the MU MMG class MMG-P, it is mainly the distal muscle surface deformation. For the MU MMG of MMG-M type the effects of deformation within the proximal and distal muscle surfaces overlap. The model has been verified with experimental recordings, and its responses are consistent and adequate in comparison to the experimental data.

Model-generated decomposition of unfused tetani of motor units evoked by random stimulation.

Unfused tetani of motor units (MUs) evoked by stimulation at variable interpulse intervals at mean frequencies of 20, 25, 33, 40 and 50Hz were studied using ten functionally isolated fast-type MUs from the medial gastrocnemius muscle of adult Wistar rats. A previously proposed algorithm and computer program for mathematical decomposition of unfused tetani into a series of twitches, representing responses to individual pulses, were used. Analysis of the parameters of the decomposed twitches showed considerable variability in force of successive contractions. These twitches were extremely variable with up to 2-fold higher forces and longer contraction times than a single twitch evoked by one stimulus. However, when the stimulation frequency was decreased, the decomposed twitches became similar to the single twitch with respect to amplitude and contraction time. It was found that the basic contractile parameters of decomposed twitches could be predicted with high accuracy on the basis of the tetanus force level at which the next contraction begins. This analysis of the parameters of decomposed twitches demonstrated that the contractile responses of the muscle fibers to successive action potentials generated by motoneurons are highly variable and depend on the previous MU state.