Contractions Of Motor Units Research Articles

Optimization and comparison of two methods for spike train estimation in an unfused tetanic contraction of low threshold motor units

BackgroundRecent findings have shown that imaging voluntarily activated motor units (MUs) by decomposing ultrasound-based displacement images provides estimates of unfused tetanic signals evoked by spinal motoneurons’ neural discharges (spikes). Two methods have been suggested to estimate its spike trains: band-pass filter (BPM) and Haar wavelet transform (HWM). However, the methods’ optimal parameters and which method performs the best are unknown. This study will answer these questions. MethodHWM and BPM were optimized using simulations. Their performance was evaluated based on simulations and 21 experimental datasets, considering their rate of agreement, spike offset, and spike offset variability to the simulated or experimental spikes. ResultsA range of parameter sets that resulted in the highest possible agreement with simulated spikes was provided. Both methods highly agreed with simulated and experimental spikes, but HWM was a better spike estimation method than BPM because it had a higher agreement, less bias, and less variation (p < 0.001). ConclusionsThe optimized HWM will be an important contributor to further developing the identification and analysis of MUs using imaging, providing indirect access to the neural drive of the spinal cord to the muscle by the unfused tetanic signals.

Estimation of contractile parameters of successive twitches in unfused tetanic contractions of single motor units – A proof-of-concept study using ultrafast ultrasound imaging in vivo

During a voluntary contraction, motor units (MUs) fire a train of action potentials, causing summation of the twitch forces, resulting in fused or unfused tetanus. Twitches have been important in studying whole-muscle contractile properties and differentiation between MU types. However, there are still knowledge gaps concerning the voluntary force generation mechanisms. Current methods rely on the spike-triggered averaging technique, which cannot track changes in successive twitches’ properties in response to individual neural firings. This study proposes a method that estimates successive twitches contractile parameters of single MUs during low force voluntary isometric contractions in human biceps brachii. We used a previously developed ultrafast ultrasound imaging method to estimate unfused tetanic activity signals of single MUs. A twitch decomposition model was used to decompose unfused tetanic activity signals into individual twitches. This study found that the contractile parameters varied within and across MUs. There was an association between the inter-spike interval and the contraction time (r = 0.49,p < 0.001) and the half-relaxation time (r = 0.58,p < 0.001), respectively. The method shows the proof-of-concept to study MU contractile properties of individual twitches in vivo, which can provide further insights into the force generation mechanisms of voluntary contractions and response to individual neural discharges.

Kinematics of individual muscle units in natural contractions measured in vivo using ultrafast ultrasound

Objective. The study of human neuromechanical control at the motor unit (MU) level has predominantly focussed on electrical activity and force generation, whilst the link between these, i.e. the muscle deformation, has not been widely studied. To address this gap, we analysed the kinematics of muscle units in natural contractions. Approach. We combined high-density surface electromyography (HDsEMG) and ultrafast ultrasound (US) recordings, at 1000 frames per second, from the tibialis anterior muscle to measure the motion of the muscular tissue caused by individual MU contractions. The MU discharge times were identified online by decomposition of the HDsEMG and provided as biofeedback to 12 subjects who were instructed to keep the MU active at the minimum discharge rate (9.8 ± 4.7 pulses per second; force less than 10% of the maximum). The series of discharge times were used to identify the velocity maps associated with 51 single muscle unit movements with high spatio-temporal precision, by a novel processing method on the concurrently recorded US images. From the individual MU velocity maps, we estimated the region of movement, the duration of the motion, the contraction time, and the excitation–contraction (E–C) coupling delay. Main results. Individual muscle unit motions could be reliably identified from the velocity maps in 10 out of 12 subjects. The duration of the motion, total contraction time, and E–C coupling were 17.9 5.3 ms, 56.6 8.4 ms, and 3.8 3.0 ms (n = 390 across ten participants). The experimental measures also provided the first evidence of muscle unit twisting during voluntary contractions and MU territories with distinct split regions. Significance. The proposed method allows for the study of kinematics of individual MU twitches during natural contractions. The described measurements and characterisations open new avenues for the study of neuromechanics in healthy and pathological conditions.

Shaky hands are a part of motor neuron disease phenotype: clinical and electrophysiological study of 77 patients.

In the sharp contrast with the existing literature, we frequently observe minipolymyoclonus, tremor and pseudodystonic thumb posturing in patients with motor neuron disease. We conducted a clinical and electrophysiological study to describe phenomenology, prevalence and pathophysiology of involuntary movements in motor neuron disease. We included 77 consecutive patients. Involuntary movements were assessed at rest and on action. Patients were videotaped. Arm muscle tone, power and deep tendon reflexes were evaluated. Accelerometry with electromyography was recorded in a subset of patients. Involuntary movements were observed in 68.9% of patients and could be separated into rest minipolymyoclonus, thumb tremor, pseudodystonic thumb posture, action minipolymyoclonus, and action tremor. One-third of patients reported negative impact of involuntary movements on hand use. Logistic regression showed that rest minipolymyoclonus and thumb tremor were more likely to occur in patients with more prominent distal muscle weakness and less spasticity. Similarly, action involuntary movements were more likely to appear in weaker patients. Patients with brisk tendon reflexes were more likely to display action tremor than action minipolymyoclonus. Action tremor was characterized by accelerometer and corresponding electromyography peak frequency, which decreased with mass loading, suggesting a mechanical-reflex tremor. Involuntary movements are common, but poorly recognized feature of motor neuron disease that may add to functional impairment. Results of our study suggest that involuntary movements are likely of peripheral origin, with a non-fused contraction of enlarged motor units being a common driving mechanism. Minipolymyoclonus appears if no synchronization of motor units occurs. When synchronization occurs via stretch reflex, mechanical-reflex tremor is generated.

A small dose of succinylcholine versus de-fasciculating dose of pancuronium for the prevention of succinylcholine-induced side effects in surgical patients: A prospective cohort study

BackgroundThe onset of paralysis by succinylcholine is usually signaled by visible motor unit contractions called fasciculation. Even though it is a short-acting muscle relaxant of choice that facilitates endotracheal intubation, it has some side effects. The most common side effects are fasciculation and post-operative myalgias which are unpleasant experiences for the patient lasting 24–48 h after surgery. The current study aimed to compare the effect of a mini-dose of succinylcholine versus a de-fasciculating dose of pancuronium for the prevention of succinylcholine-induced side effects in surgical patients. MethodsThis is a prospective cohort study that recruited 96 patients who underwent elective surgery. Patients were grouped into three study arms: 1. 0.01mg/kg pancuronium, 2. 0.2mg/kg succinylcholine, and 3. with no pretreatment. Data were analyzed using one-way analysis of variance (ANOVA) with post hoc analysis for multiple tests and non-normally distributed data were analyzed using the Kuruska-Wallih H rank test. P-values < 0.05 were considered as statistically significant. ResultsThe total incidence of succinylcholine-induced fasciculation was 17 (53.1%), 8(25%), and 30 (93.8%) in pancuronium, mini-dose of succinylcholine pretreatment and unexposed groups, respectively (p<0.001). The severity of fasciculation was reduced more in mini-dose succinylcholine than in the other two groups. The total incidence of myalgia was 6(18.8%), 3(9.4%), and 9(28.1%) in pancuronium, mini-dose of succinylcholine, and unexposed groups, respectively (p=0.158). There was no difference in the severity of myalgia between groups (p=0.161). There was no statistically significant difference in the incidence and severity of succinylcholine-induced postoperative myalgia among the three groups. ConclusionPretreatment with a mini-dose of succinylcholine might reduce the incidence and severity of succinylcholine-induced fasciculation in patients undergoing elective surgery. In this study, pre-treatment with a mini-dose of succinylcholine or pancuronium did not demonstrate an effect on the incidence or severity of succinylcholine-induced myalgia.

Microendoscopy detects altered muscular contractile dynamics in a mouse model of amyotrophic lateral sclerosis

Amyotrophic lateral sclerosis (ALS) is a fatal disease involving motor neuron degeneration. Effective diagnosis of ALS and quantitative monitoring of its progression are crucial to the success of clinical trials. Second harmonic generation (SHG) microendoscopy is an emerging technology for imaging single motor unit contractions. To assess the potential value of microendoscopy for diagnosing and tracking ALS, we monitored motor unit dynamics in a B6.SOD1G93A mouse model of ALS for several weeks. Prior to overt symptoms, muscle twitch rise and relaxation time constants both increased, consistent with a loss of fast-fatigable motor units. These effects became more pronounced with disease progression, consistent with the death of fast fatigue-resistant motor units and superior survival of slow motor units. From these measurements we constructed a physiological metric that reflects the changing distributions of measured motor unit time constants and effectively diagnoses mice before symptomatic onset and tracks disease state. These results indicate that SHG microendoscopy provides a means for developing a quantitative, physiologic characterization of ALS progression.

O-24 MRI detection of human motor unit fasciculation in Amyotrophic Lateral Sclerosis

Objective To develop a clinically applicable non-invasive method for detecting motor unit fasciculation in patients with Amyotrophic Lateral Sclerosis (ALS). Methods The lower limbs of 6 healthy controls and 4 patients with confirmed ALS were scanned using a novel diffusion weighted MRI protocol sensitive to micrometer-scale movement of skeletal muscle. Motor unit activity was assessed at rest and during electrical motor nerve stimulation time-locked to scanner acquisition. Results Incremental stimulation revealed a reproducible pattern of overlapping signal voids with dimensions and temporal profiles consistent with the contraction of single motor units. Patients with ALS showed a significantly higher rate of spontaneous motor unit fasciculation at rest (mean 99.1/min, range 25.7–161 in patients vs 7.7/min, range 4.3–9.7 in controls, p Conclusions This study is the first use of imaging to detect fasciculation in ALS patients. The technique is quick to perform and entirely pain-free. The ability to detect changes in motor unit function which precede motor unit degeneration may allow earlier diagnosis and recruitment to clinical trials.

P31-F Muscle ultrasound improves fasciculation detection in some muscle in patients with amyotrophic lateral sclerosis

Background Fasciculations represent brief spontaneous or triggered contractions of motor units that can be seen as flickering movements under the skin and represent a valuable sign of lower motor neuron lesion. It is sometimes difficult to detect fasciculations clinically and/or electromyografically (EMG). Muscle ultrasound (MUS) is a promising tool that can aid to the better detection of fasciculations. Materials and methods 12 patients (4 females, mean age 68.75 ± 12.04, mean disease duration 4.25 ± 2.09) diagnosed with amyotrophic lateral sclerosis were included in this pilot study. EMG needle was inserted under US guidance into right biceps brachii (BB), first dorsal interousseous (FDI) and rectus femoris (RF) muscles. Location of needle tip was marked and remained stationary during 90 s of recording. Two experienced neurophysiologists independently counted the number of fasciculations from the EMG and MUS recordings. Sperman’s ρ was used to assess interrater reliability. Wilcoxon sign-ranked test was used to compare the number of fasciculations detected by the use of EMG vs. MUS. Results There interrater reliability was excellent (ρ = 0.985, p Conclusions MUS seems to be superior to EMG in detecting and quantifying fasciculations in some, but not all muscles. Further studies are needed to standardize the use of MUS in detecting fasciculations in neurological diseases.

Transitory force decrease following a sudden reduction in stimulation frequency in motor units of rat medial gastrocnemius

Effects of a sudden decrease in the stimulation frequency for motor unit force were studied in rat medial gastrocnemius. For 161 functionally isolated single motor units of three types (S, FR, FF), unfused tetanic contractions were evoked by three-phase trains of stimuli (low–high–low frequency). The course of the tetanus at the onset of the third phase of the force recording was analyzed in tetani with variable fusion degree. For 78 units within the third phase of tetanus, a transitory force decrease to a level lower than in the first phase (identical frequency), was observed. This phenomenon was more frequent for fast fatigue resistant (65.9%) than for fast fatigable and slow motor units (27.1% and 35.5%, respectively). Moreover, the force decrease was strongest for fast resistant motor units (up to 36.5%) and when contractions evoked at variable frequencies of stimulation were compared, the highest amplitudes of the studied force decrease were noted for middle-fused tetani (0.50–0.90). A new phenomenon of transitory force decrease in tetanic contractions of motor units with a decrease in stimulation frequency was found. Most probably, the phenomenon is dependent on disturbances in the force transmission by collagen surrounding active muscles fibers.

Functional magnetic resonance imaging of human motor unit fasciculation in amyotrophic lateral sclerosis.

A novel diffusion-weighted magnetic resonance imaging protocol sensitive to contraction of individual skeletal motor units was developed. We applied this technique to the lower limb muscles of 4 patients with confirmed amyotrophic lateral sclerosis (ALS) and 6 healthy controls. A 3-minute scan revealed florid fasciculation in ALS patients, involving both superficial and deep muscles, and at a frequency higher than in healthy controls. This novel imaging technique reveals hitherto unobtainable information on human motor unit structure and function, which may allow earlier diagnosis and recruitment to clinical trials. ANN NEUROL 2019;85:455-459.

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.

The neural response properties and cortical organization of a rapidly adapting muscle sensory group response that overlaps with the frequencies that elicit the kinesthetic illusion.

Kinesthesia is the sense of limb movement. It is fundamental to efficient motor control, yet its neurophysiological components remain poorly understood. The contributions of primary muscle spindles and cutaneous afferents to the kinesthetic sense have been well studied; however, potential contributions from muscle sensory group responses that are different than the muscle spindles have not been ruled out. Electrophysiological recordings in peripheral nerves and brains of male Sprague Dawley rats with a degloved forelimb preparation provide evidence of a rapidly adapting muscle sensory group response that overlaps with vibratory inputs known to generate illusionary perceptions of limb movement in humans (kinesthetic illusion). This group was characteristically distinct from type Ia muscle spindle fibers, the receptor historically attributed to limb movement sensation, suggesting that type Ia muscle spindle fibers may not be the sole carrier of kinesthetic information. The sensory-neural structure of muscles is complex and there are a number of possible sources for this response group; with Golgi tendon organs being the most likely candidate. The rapidly adapting muscle sensory group response projected to proprioceptive brain regions, the rodent homolog of cortical area 3a and the second somatosensory area (S2), with similar adaption and frequency response profiles between the brain and peripheral nerves. Their representational organization was muscle-specific (myocentric) and magnified for proximal and multi-articulate limb joints. Projection to proprioceptive brain areas, myocentric representational magnification of muscles prone to movement error, overlap with illusionary vibrational input, and resonant frequencies of volitional motor unit contraction suggest that this group response may be involved with limb movement processing.

Capillary endothelial cells as coordinators of skeletal muscle blood flow during active hyperemia.

In this invited review, we explore the burgeoning possibility of capillary endothelial cells as coordinators of skeletal muscle blood flow in response to muscle contraction. The idea that the capillary is an active vascular unit in skeletal muscle microcirculation starkly diverges from the traditional dogma that seats arterioles as the central controllers of blood flow during exercise. This review aims to incite discussion as we revisit and rethink the role of capillary endothelial cells in skeletal muscle. We discuss the potential for a mismatch in the architectural relationships between the arteriolar microvasculature and contracting motor units that would negate consistent communication between them. We review the data from the past two decades demonstrating that capillaries are ideally located architecturally to communicate with skeletal muscle fibers and are mechanistically capable of signaling upstream arterioles that control their own perfusion. We show that the orchestration of a coordinated vascular response necessary to support active skeletal muscle fibers cannot be achieved by the arterioles, but rather it is the capillaries that drive the blood flow response to muscle contraction. Thus, capillaries need to be seriously considered as critical in the coordination of skeletal muscle blood flow during active hyperemia.

A General Mathematical Algorithm for Predicting the Course of Unfused Tetanic Contractions of Motor Units in Rat Muscle.

An unfused tetanus of a motor unit (MU) evoked by a train of pulses at variable interpulse intervals is the sum of non-equal twitch-like responses to these stimuli. A tool for a precise prediction of these successive contractions for MUs of different physiological types with different contractile properties is crucial for modeling the whole muscle behavior during various types of activity. The aim of this paper is to develop such a general mathematical algorithm for the MUs of the medial gastrocnemius muscle of rats. For this purpose, tetanic curves recorded for 30 MUs (10 slow, 10 fast fatigue-resistant and 10 fast fatigable) were mathematically decomposed into twitch-like contractions. Each contraction was modeled by the previously proposed 6-parameter analytical function, and the analysis of these six parameters allowed us to develop a prediction algorithm based on the following input data: parameters of the initial twitch, the maximum force of a MU and the series of pulses. Linear relationship was found between the normalized amplitudes of the successive contractions and the remainder between the actual force levels at which the contraction started and the maximum tetanic force. The normalization was made according to the amplitude of the first decomposed twitch. However, the respective approximation lines had different specific angles with respect to the ordinate. These angles had different and non-overlapping ranges for slow and fast MUs. A sensitivity analysis concerning this slope was performed and the dependence between the angles and the maximal fused tetanic force normalized to the amplitude of the first contraction was approximated by a power function. The normalized MU contraction and half-relaxation times were approximated by linear functions depending on the normalized actual force levels at which each contraction starts. The normalization was made according to the contraction time of the first contraction. The actual force levels were calculated initially from the recorded tetanic curves and subsequently from the modeled curves obtained from the summation of all models of the preceding contractions (the so called “full prediction”). The preciseness of the prediction was verified by two coefficients estimating the error between the modeled and the experimentally recorded curves. The proposed approach was tested for 30 MUs from the database and for three additional MUs, not included in the initial set. It was concluded that this general algorithm can be successfully used for modeling of a unfused tetanus course of a single MU of fast and slow type.

Prolonged activity evokes potentiation and the "sag" phenomenon in slow motor units of rat soleus.

Slow motor units (MUs) have no sag in their unfused tetani. This study in anesthetized rats shows that the sag can be observed in slow soleus MUs after prolonged activity. Twitches and unfused tetanic contractions were recorded from male (n=35) and female (n=39) MUs before and after the four minutes of the fatigue test (trains of 13 pulses at 40 Hz repeated every second). After this activity twitch contractions potentiated and a shift in the steep part of the force-frequency curve towards lower frequencies was observed in both sexes. Initially no sag was visible in unfused tetani, but after the fatigue test the phenomenon was observed in 77% of male, while in 13% of female MUs, the result consistent with the previously reported higher content of IIa myosin and faster contraction of MUs in male soleus. The decomposition of tetani with sag into trains of twitch-shape responses to consecutive stimuli revealed higher forces of initial decomposed twitches than later. The revealed alterations the force development due to long-lasting activation of slow MUs were sex-related and more pronounced in male soleus.

In Vivo Imaging of Human Sarcomere Twitch Dynamics in Individual Motor Units

Motor units comprise a pre-synaptic motor neuron and multiple post-synaptic muscle fibers. Many movement disorders disrupt motor unit contractile dynamics and the structure of sarcomeres, skeletal muscle's contractile units. Despite the motor unit's centrality to neuromuscular physiology, no extant technology can image sarcomere twitch dynamics in live humans. We created a wearable microscope equipped with a microendoscope for minimally invasive observation of sarcomere lengths and contractile dynamics in any major skeletal muscle. By electrically stimulating twitches via the microendoscope and visualizing the sarcomere displacements, we monitored single motor unit contractions in soleus and vastus lateralis muscles of healthy individuals. Control experiments verified that these evoked twitches involved neuromuscular transmission and faithfully reported muscle force generation. In post-stroke patients with spasticity of the biceps brachii, we found involuntary microscopic contractions and sarcomere length abnormalities. The wearable microscope facilitates exploration of many basic and disease-related neuromuscular phenomena never visualized before in live humans.

Decomposition of motor unit tetanic contractions of rat soleus muscle: Differences between males and females

Mathematical decomposition of tetanic contractions of slow motor units (MUs) of the rat heterogeneous medial gastrocnemius muscle revealed immense variability of twitch-shape responses to successive pulses, contrary to results obtained for fast MUs. The aim of this study in rat soleus muscle, almost exclusively composed of slow MUs, was to reveal whether such variability of twitch-shape decomposed components was a common property of slow MUs in the two studied muscles, and whether ranges of the force amplitude or time parameters of these decomposed twitches showed sex differences. Unfused tetanic contractions evoked by stimulation at variable interpulse intervals were analyzed for 10 MUs of males and 10 MUs of females. Significantly higher variability between parameters of the decomposed responses was observed for male soleus MUs, as the mean ratio of forces of the strongest decomposed twitch and the first (the weakest) decomposed twitch amounted to 3.8 for males and 2.8 for females. The ratios of the contraction times of the longest decomposed to the first twitch were much more similar between male and female MUs, 2.6 and 2.9, respectively. Consequently, the mean ratio of the force–time area for the strongest decomposed to the first twitch was much bigger in male than female MUs (7.35 vs. 5.07, respectively). Our observations indicate that high variability of responses to successive stimuli is a general property of slow MUs in different rat muscles, but the mechanisms of summation of individual twitches into tetanic contractions of MUs are not identical for male and female rats.

Estimation of the error between experimental tetanic force curves of MUs of rat medial gastrocnemius muscle and their models by summation of equal successive contractions

More accurate muscle models require appropriate modelling of individual twitches of motor units (MUs) and their unfused tetanic contractions. It was shown in our previous papers, using a few MUs, that modelling of unfused tetanic force curves by summation of equal twitches is not accurate, especially for slow MUs. The aim of this study was to evaluate this inaccuracy using a statistical number of MUs of the rat medial gastrocnemius muscle (15 of slow, 15 of fast resistant and 15 of fast fatigable type). Tetanic contractions were evoked by trains of 41 stimuli at random interpulse intervals and different mean frequencies, resembling discharge patterns observed during natural muscle activity. The tetanic curves were calculated by the summation of equal twitches according to the respective experimental patterns. The previously described 6-parameter analytical function for twitch modelling was used. Comparisons between the experimental and the modelled curves were made using two coefficients: the fit coefficient and the area coefficient. The errors between modelled and experimental tetanic forces were substantially different between the three MU types. The error was the most significant for slow MUs, which develop much higher forces in real contractions than could be predicted based on the summation of equal twitches, while the smallest error was observed for FF MUs – their recorded tetanic forces were similar to those predicted by modelling. The obtained results indicate the importance of the inclusion of the type-specific non-linearity in the summation of successive twitch-like contractions of MUs in order to increase the reliability of modelling skeletal muscle force.

Physiological consequences of doublet discharges on motoneuronal firing and motor unit force.

The double discharges are observed at the onset of contractions of mammalian motor units (MUs), especially during their recruitment to strong or fast movements. Doublets lead to MU force increase and improve ability of muscles to maintain high force during prolonged contractions. In this review we discuss an ability to produce doublets by fast and slow motoneurons (MNs), their influence on the course of action potential afterhyperpolarization (AHP) as well as its role in modulation of the initial stage of the firing pattern of MNs. In conclusion, a generation of doublets is an important strategy of motor control, responsible for fitting the motoneuronal firing rate to the optimal for MUs at the start of their contraction, necessary for increment of muscle force.

An Approach for Simulation of the Muscle Force Modeling It by Summation of Motor Unit Contraction Forces

Muscle force is due to the cumulative effect of repetitively contracting motor units (MUs). To simulate the contribution of each MU to whole muscle force, an approach implemented in a novel computer program is proposed. The individual contraction of an MU (the twitch) is modeled by a 6-parameter analytical function previously proposed; the force of one MU is a sum of its contractions due to an applied stimulation pattern, and the muscle force is the sum of the active MUs. The number of MUs, the number of slow, fast-fatigue-resistant, and fast-fatigable MUs, and their six parameters as well as a file with stimulation patterns for each MU are inputs for the developed software. Different muscles and different firing patterns can be simulated changing the input data. The functionality of the program is illustrated with a model consisting of 30 MUs of rat medial gastrocnemius muscle. The twitches of these MUs were experimentally measured and modeled. The forces of the MUs and of the whole muscle were simulated using different stimulation patterns that included different regular, irregular, synchronous, and asynchronous firing patterns of MUs. The size principle of MUs for recruitment and derecruitment was also demonstrated using different stimulation paradigms.