Human Motor Units Research Articles

Inducing post-activation potentiation alters human motor unit behavior of the elbow extensors during unconstrained velocity contractions.

The purpose was to test whether inducing post-activation potentiation (PAP) altered motor unit (MU) activity during dynamic isotonic contractions. From 12 participants (3 females), 39 MUs were recorded from the anconeus (n=31) and lateral triceps brachii (n=8) with fine-wire electrodes during elbow extensions at 50 and 75% of peak power with, and without PAP. To induce PAP participants produced a 2s ramp conditioning contraction (CC) up to maximal isometric elbow extension with a 3s hold. Following the CC (~2s), independent electrical stimulation to the triceps and anconeus showed twitch torques were potentiated by 84 and 66%, respectively, (both p<0.001). Compared to baseline (i.e., without PAP), at both intensities (50 and 75%) PAP increased MU recruitment thresholds (40% and 80%, p<0.001) with lowered mean MU rates (-20 and -26%), and instantaneous rates at recruitment threshold (-26 and -25%) (all p<0.001). Firing rates increased 20% (p<0.001) from 50 to 75% power, but rates during potentiated contractions targeting 75% were lower than baseline at 50% (-10%, p<0.001). Dynamic contractions provide a more functional paradigm to assess MU activity with PAP and showed larger effects across a wider range of contractile intensities compared to previously described isometric tasks. Findings indicate that peripheral feedback from the potentiated muscle is likely not the primary mechanism in modifying MU behaviors as changes occurred at recruitment which is relatively insensitive to afferent feedback. Therefore, MU activity during dynamic contractions is responsive to activation history force potentiation and can make compensatory adjustments to optimize contractile output.

Antagonism of 5-HT2 receptors attenuates self-sustained firing of human motor units.

5-HT2 receptors on motoneurones play a critical role in facilitating persistent inward currents (PICs). Although facilitation of PICs can enhance self-sustained firing after periods of excitation, the relationship between 5-HT2 receptor activity and self-sustained firing in human motor units (MUs) has not been resolved. MU activity was assessed from the tibialis anterior of 10 healthy adults (24.9 ± 2.8 years) during two contraction protocols. Both protocols featured steady-state isometric contractions with constant descending drive to the motoneurone pool. However, one protocol also included an additional phase of superimposed descending drive. Adding and then removing descending drive in the middle of steady-state contractions altered MU firing behaviour across the motor pool, where newly recruited units in the superimposed phase were unable to switch off (P=0.0002), and units recruited prior to additional descending drive reduced their discharge rates (P<0.0001, difference in estimated marginal means (∆)=2.24 pulses/s). The 5-HT2 receptor antagonist, cyproheptadine, was then administered to determine whether changes in MU firing were mediated by serotonergic mechanisms. 5-HT2 receptor antagonism caused reductions in MU discharge rate (P<0.001, ∆=1.65 pulses/s), recruitment threshold (P=0.00112, ∆=1.09% maximal voluntary contraction) and self-sustained firing duration (P<0.0001, ∆=1.77s) after the additional descending drive was removed in the middle of the steady-state contraction. These findings indicate that serotonergic neuromodulation plays a key role in facilitating discharge and self-sustained firing of human motoneurones, where adaptive changes in MU recruitment must occur to meet the demands of the contraction. KEY POINTS: Animal and cellular preparations indicate that somato-dendritic 5-HT2 receptors regulate the intrinsic excitability of motoneurones. 5-HT2 receptor antagonism reduces estimates of persistent inward currents in motoneurones, which contribute to self-sustained firing when synaptic inputs are reduced or removed. This human study employed a contraction task that slowly increased (and then removed) the additional descending drive in the middle of a steady-state contraction where marked self-sustained firing occurred when the descending drive was removed. 5-HT2 receptor antagonism caused widespread reductions in motor unit (MU) discharge rates during contractions, which was accompanied by reduced recruitment threshold and attenuation of self-sustained firing duration after the removal of the additional descending drive to motoneurones. These findings support the role that serotonergic neuromodulation is a key facilitator of MU discharge and self-sustained firing of human motoneurones, where adaptative changes in MU recruitment must occur to meet the demands of the contraction.

Postinhibitory excitation in motoneurons can be facilitated by hyperpolarization-activated inward currents: A simulation study.

Postinhibitory excitation is a transient overshoot of a neuron's baseline firing rate following an inhibitory stimulus and can be observed in vivo in human motoneurons. However, the biophysical origin of this phenomenon is still unknown and both reflex pathways and intrinsic motoneuron properties have been proposed. We hypothesized that postinhibitory excitation in motoneurons can be facilitated by hyperpolarization-activated inward currents (h-currents). Using an electrical circuit model, we investigated how h-currents can modulate the postinhibitory response of motoneurons. Further, we analyzed the spike trains of human motor units from the tibialis anterior muscle during reciprocal inhibition. The simulations revealed that the activation of h-currents by an inhibitory postsynaptic potential can cause a short-term increase in a motoneuron's firing probability. This result suggests that the neuron can be excited by an inhibitory stimulus. In detail, the modulation of the firing probability depends on the time delay between the inhibitory stimulus and the previous action potential. Further, the postinhibitory excitation's strength correlates with the inhibitory stimulus's amplitude and is negatively correlated with the baseline firing rate as well as the level of input noise. Hallmarks of h-current activity, as identified from the modeling study, were found in 50% of the human motor units that showed postinhibitory excitation. This study suggests that h-currents can facilitate postinhibitory excitation and act as a modulatory system to increase motoneuron excitability after a strong inhibition.

Acute and prolonged competing effects of activation history on human motor unit firing rates during contractile impairment and recovery

AbstractThe purpose of this study was to investigate the effect of inducing post‐activation potentiation (PAP) during prolonged low‐frequency force depression (PLFFD) on motor unit (MU) firing rates. In 10 participants, grouped firing rates of 3027 MUs from the tibialis anterior were recorded with tungsten microelectrodes. Baseline MU firing rates at 25% isometric maximal voluntary contraction (MVC) were ∼14 Hz. A 1 min dorsiflexion MVC reduced torque and maximal MU firing rates (36 Hz) by 49% and 52%, respectively. Following task completion, firing rates at 25% of baseline MVC torque and torque in response to electrically evoked (single twitch, 10 Hz and 50 Hz) stimulation were assessed before and after a 5 s MVC (to induce PAP) every 10 min for 60 min. From 10 to 60 min after task completion, the torque ratios (twitch:50 Hz and 10:50 Hz) were depressed (∼30%) relative to baseline (P &lt; 0.001), indicating PLFFD; and firing rates were higher by ∼15% relative to baseline (P &lt; 0.001). This occurred despite recovery of MVC rates (∼99%) and torque (∼95%) by 10 min (P &gt; 0.3). Inducing PAP during PLFFD increased both low to high torque ratios (twitch and 10:50 Hz) by ∼200% and ∼135%, respectively (P &lt; 0.001) and firing rates were ∼18% lower relative to PLFFD rates (P &lt; 0.001), despite a speeding of evoked contractile properties (P = 0.001). Thus, firing rates appear strongly matched to alterations in torque, rather than contractile speed when modified by contractile history, and lower rates during PAP may be a mechanism to mitigate effects of PLFFD. The effect of activation history on contractile function demonstrates acute compensatory responses of motoneuron output. imageKey points Prolonged low frequency force depression (PLFFD) following a sustained 1 min isometric maximal voluntary contraction causes an increase in submaximal mean motor unit (MU) firing rates. Inducing post‐activation potentiation (PAP) during PLFFD, however, causes a reduction in mean submaximal MU firing rates to a level below those at baseline. The mean firing rate reduction during PAP occurs despite a speeding of evoked contractile properties and thus firing rates are more strongly matched to alterations in torque, rather than contractile speed when modified by various contractile histories. The reductions in firing rates during PAP may mitigate the effects of PLFFD during voluntary contractions. These results demonstrate that firing rates are highly responsive to opposing influences on the contractile state and can make rapid compensatory rate adjustments dependent on the active state of the muscle.

Menstrual Cycle Associated Alteration of Vastus Lateralis Motor Unit Function

BackgroundEstrogen and progesterone are the primary female sex hormones and have net excitatory and inhibitory effects, respectively, on neuronal function. Fluctuating concentrations across the menstrual cycle has led to several lines of research in relation to neuromuscular function and performance; however evidence from animal and cell culture models has yet to be demonstrated in human motor units coupled with quantification of circulating hormones. Intramuscular electromyography was used to record motor unit potentials and corresponding motor unit potential trains from the vastus lateralis of nine eumenorrheic females during the early follicular, ovulation and mid luteal phases of the menstrual cycle, alongside assessments of neuromuscular performance. Multi-level regression models were applied to explore effects of time and of contraction level. Statistical significance was accepted as p < 0.05.ResultsKnee extensor maximum voluntary contraction, jump power, force steadiness, and balance did not differ across the menstrual phases (all p > 0.4). Firing rate of low threshold motor units (10% maximum voluntary contraction) was lower during the ovulation and mid luteal phases (β = − 0.82 Hz, p < 0.001), with no difference in motor unit potentials analysed from 25% maximum voluntary contraction contractions. Motor unit potentials were more complex during ovulation and mid luteal phase (p < 0.03), with no change in neuromuscular junction transmission instability (p > 0.3).ConclusionsAssessments of neuromuscular performance did not differ across the menstrual cycle. The suppression of low threshold motor unit firing rate during periods of increased progesterone may suggest a potential inhibitory effect and an alteration of recruitment strategy; however this had no discernible effect on performance. These findings highlight contraction level-dependent modulation of vastus lateralis motor unit function over the eumenorrheic cycle, occurring independently of measures of performance.

Simulating progressive motor neuron degeneration and collateral reinnervation in motor neuron diseases using a dynamic muscle model based on human single motor unit recordings

Objective. To simulate progressive motor neuron loss and collateral reinnervation in motor neuron diseases (MNDs) by developing a dynamic muscle model based on human single motor unit (MU) surface-electromyography (EMG) recordings. Approach. Single MU potentials recorded with high-density surface-EMG from thenar muscles formed the basic building blocks of the model. From the baseline MU pool innervating a muscle, progressive MU loss was simulated by removal of MUs, one-by-one. These removed MUs underwent collateral reinnervation with scenarios varying from 0% to 100%. These scenarios were based on a geometric variable, reflecting the overlap in MU territories using the spatiotemporal profiles of single MUs and a variable reflecting the efficacy of the reinnervation process. For validation, we tailored the model to generate compound muscle action potential (CMAP) scans, which is a promising surface-EMG method for monitoring MND patients. Selected scenarios for reinnervation that matched observed MU enlargements were used to validate the model by comparing markers (including the maximum CMAP and a motor unit number estimate (MUNE)) derived from simulated and recorded CMAP scans in a cohort of 49 MND patients and 22 age-matched healthy controls. Main results. The maximum CMAP at baseline was 8.3 mV (5th–95th percentile: 4.6 mV–11.8 mV). Phase cancellation caused an amplitude drop of 38.9% (5th–95th percentile, 33.0%–45.7%). To match observations, the geometric variable had to be set at 40% and the efficacy variable at 60%–70%. The Δ maximum CMAP between recorded and simulated CMAP scans as a function of fitted MUNE was −0.4 mV (5th–95th percentile = −4.0 – +2.4 mV). Significance. The dynamic muscle model could be used as a platform to train personnel in applying surface-EMG methods prior to their use in clinical care and trials. Moreover, the model may pave the way to compare biomarkers more efficiently, without directly posing unnecessary burden on patients.

Age-related alterations in human motor units properties and in muscle innervation profile

Introduction: Mounting evidence suggests that motoneuron and neuromuscular junction (NMJ) degeneration, muscle fiber denervation and loss of motor units contribute to muscle wasting and weakness in old age. However, these aspects are still poorly investigated in humans. This study aimed at comparing the neuromuscular system integrity and function of young and older adults, with particular focus on NMJ and motor unit potential (MUP) characteristics. We hypothesized that older individuals would present altered electrophysiological properties and innervation profile. Methods: We recruited 55 healthy young individuals (YI) (aged 26.1±4.5 yr; 50% females) and 88 older individuals (OI) (aged 75.9±4.7 yr; 55% females). Muscle force was evaluated by isometric dynamometry, muscle morphology by ultrasound and neuromuscular excitability by compound motor unit action potential (CMAP). MUP properties and motor unit number estimate (iMUNE) were obtained by intramuscular electromyography (iEMG). In a sub-cohort of individuals (n=16 YI and n=34 OI), vastus lateralis (VL) muscle biopsies were obtained. We assessed denervation status by scoring neural cell adhesion molecule (NCAM) positive fibers. Generalized linear mixed models were carried out for iEMG data, while unpaired t-tests were carried out for all the other parameters. Results: OI showed alterations in muscle morphology when compared to YI, evident as smaller whole quadriceps cross-sectional area (p&lt;0.001) and VL fiber diameter (p&lt;0.01), together with smaller VL pennation angles (p&lt;0.001) and fascicle lengths (p&lt;0.05), resulting in an increased ultrasound sarcopenia index (p=0.001). Muscle force (p&lt;0.001) and CMAP (p&lt;0.001) were also largely impaired in OI, suggesting that muscle function and neuromuscular excitability are considerably affected by aging. iEMG analysis revealed changes in MUP turns (p&lt;0.001), near fiber (NF) MUP duration (p&lt;0.001) and NF count (p&lt;0.001) in OI, pointing towards an increased MUP complexity, in absence of sex differences. The iMUNE was reduced (p&lt;0.001) and the NMJ transmission stability was significantly altered in OI, as suggested by elevated NF jiggle (p&lt;0.001) and segmented jitter (p&lt;0.05). These electrophysiological alterations seem likely due to altered muscle innervation profile, as highlighted by an increased percentage of NCAM-positive fiber (p&lt;0.001). Conclusions: This study showed that changes in muscle morphology and function, MUP properties, motor units loss and denervation status occur in older humans; these may be serve as useful markers of human neuromuscular aging. The present work was funded by PRIN project NeuAge (2017CBF8NJ_001) to MVN and the Milky Way Foundation, the Baxter Foundation, and the Li Ka Shing Foundation to HMB. This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Decoding firings of a large population of human motor units from high-density surface electromyogram in response to transcranial magnetic stimulation.

We describe a novel application of methodology for high-density surface electromyography (HDsEMG) decomposition to identify motor unit (MU) firings in response to transcranial magnetic stimulation (TMS). The method is based on the MU filter estimation from HDsEMG decomposition with convolution kernel compensation during voluntary isometric contractions and its application to contractions elicited by TMS. First, we simulated synthetic HDsEMG signals during voluntary contractions followed by simulated motor evoked potentials (MEPs) recruiting an increasing proportion of the motor pool. The estimation of MU filters from voluntary contractions and their application to elicited contractions resulted in high (>90%) precision and sensitivity of MU firings during MEPs. Subsequently, we conducted three experiments in humans. From HDsEMG recordings in first dorsal interosseous and tibialis anterior muscles, we demonstrated an increase in the number of identified MUs during MEPs evoked with increasing stimulation intensity, low variability in the MU firing latency and a proportion of MEP energy accounted for by decomposition similar to voluntary contractions. A negative relationship between the MU recruitment threshold and the number of identified MU firings was exhibited during the MEP recruitment curve, suggesting orderly MU recruitment. During isometric dorsiflexion we also showed a negative association between voluntary MU firing rate and the number of firings of the identified MUs during MEPs, suggesting a decrease in the probability of MU firing during MEPs with increased background MU firing rate. We demonstrate accurate identification of a large population of MU firings in a broad recruitment range in response to TMS via non-invasive HDsEMG recordings. KEY POINTS: Transcranial magnetic stimulation (TMS) of the scalp produces multiple descending volleys, exciting motor pools in a diffuse manner. The characteristics of a motor pool response to TMS have been previously investigated with intramuscular electromyography (EMG), but this is limited in its capacity to detect many motor units (MUs) that constitute a motor evoked potential (MEP) in response to TMS. By simulating synthetic signals with known MU firing patterns, and recording high-density EMG signals from two human muscles, we show the feasibility of identifying firings of many MUs that comprise a MEP. We demonstrate the identification of firings of a large population of MUs in the broad recruitment range, up to maximal MEP amplitude, with fewer required stimuli compared to intramuscular EMG recordings. The methodology demonstrates an emerging possibility to study responses to TMS on a level of individual MUs in a non-invasive manner.

Blockade of 5-HT2 receptors suppresses motor unit firing and estimates of persistent inward currents during voluntary muscle contraction in humans.

Serotonergic neuromodulation contributes to enhanced voluntary muscle activation. However, it is not known how the likely motoneurone receptor candidate (5-HT2 ) influences the firing rate and activation threshold of motor units (MUs) in humans. The purpose of this study was to determine whether 5-HT2 receptor activity contributes to human MU behaviour during voluntary ramped contractions of differing intensity. High-density surface EMG (HDsEMG) of the tibialis anterior was assessed during ramped isometric dorsiflexions at 10, 30, 50 and 70% of maximal voluntary contraction (MVC). MU characteristics were successfullyextracted from HDsEMG of 11 young adults (four female) pre- and post-ingestion of 8mg cyproheptadine or a placebo. Antagonism of 5-HT2 receptorscaused a reduction in MU discharge rate during steady-state muscle activation that was independent of the level of contraction intensity [P<0.001; estimated mean difference (∆)=1.06pulses/s], in addition to an increase in MU derecruitment threshold (P<0.013, ∆=1.23% MVC), without a change in force during MVC (P=0.652). A reduction in estimates of persistent inward current amplitude was observed at 10% MVC (P<0.001, ∆=0.99 Hz) and 30% MVC (P=0.003, ∆=0.75 Hz) that aligned with 5-HT changes in MU firing behaviour attributable to 5-HT2 antagonism. Overall, these findings indicate that 5-HT2 receptor activity has a role in regulating the discharge rate in populations of spinal motoneurones when performing voluntary contractions. This study provides evidence of a direct link between MU discharge properties, persistent inward current activity and 5-HT2 receptor activity in humans. KEY POINTS: Activation of 5-HT receptors on the soma and dendrites of motoneurones regulates their excitability. Previous work using chlorpromazine and cyproheptadine has demonstrated that the 5-HT2 receptor regulates motoneurone activity in humans with chronic spinal cord injury and non-injured control subjects. It is not known how the 5-HT2 receptor directly influences motor unit (MU) discharge and MU recruitment in larger populations of human motoneurones during voluntary contractions of differing intensity. Despite the absence of change in force during maximal voluntary dorsiflexions, 5-HT2 receptor antagonism caused a reduction in MU discharge rate during submaximal steady-state muscle contraction, in addition to an increase in MU derecruitment threshold, irrespective of the submaximal contraction intensity. Reductions in estimates of persistent inward currents after 5-HT2 receptor antagonism support the viewpoint that the 5-HT2 receptor plays a crucial role in regulating motor activity, whereby a persistent inward current-based mechanism is involved in regulating the excitability of human motoneurones.

A geometric approach to quantifying the neuromodulatory effects of persistent inward currents on individual motor unit discharge patterns

Objective. All motor commands flow through motoneurons, which entrain control of their innervated muscle fibers, forming a motor unit (MU). Owing to the high fidelity of action potentials within MUs, their discharge profiles detail the organization of ionotropic excitatory/inhibitory as well as metabotropic neuromodulatory commands to motoneurons. Neuromodulatory inputs (e.g. norepinephrine, serotonin) enhance motoneuron excitability and facilitate persistent inward currents (PICs). PICs introduce quantifiable properties in MU discharge profiles by augmenting depolarizing currents upon activation (i.e. PIC amplification) and facilitating discharge at lower levels of excitatory input than required for recruitment (i.e. PIC prolongation). Approach. Here, we introduce a novel geometric approach to estimate neuromodulatory and inhibitory contributions to MU discharge by exploiting discharge non-linearities introduced by PIC amplification during time-varying linear tasks. In specific, we quantify the deviation from linear discharge (‘brace height’) and the rate of change in discharge (i.e. acceleration slope, attenuation slope, angle). We further characterize these metrics on a simulated motoneuron pool with known excitatory, inhibitory, and neuromodulatory inputs and on human MUs (number of MUs; Tibialis Anterior: 1448, Medial Gastrocnemius: 2100, Soleus: 1062, First Dorsal Interosseus: 2296). Main results. In the simulated motor pool, we found brace height and attenuation slope to consistently indicate changes in neuromodulation and the pattern of inhibition (excitation–inhibition coupling), respectively, whereas the paired MU analysis (ΔF) was dependent on both neuromodulation and inhibition pattern. Furthermore, we provide estimates of these metrics in human MUs and show comparable variability in ΔF and brace height measures for MUs matched across multiple trials. Significance. Spanning both datasets, we found brace height quantification to provide an intuitive method for achieving graded estimates of neuromodulatory and inhibitory drive to individual MUs. This complements common techniques and provides an avenue for decoupling changes in the level of neuromodulatory and pattern of inhibitory motor commands.

FUS-ALS hiPSC-derived astrocytes impair human motor units through both gain-of-toxicity and loss-of-support mechanisms

BackgroundAstrocytes play a crucial, yet not fully elucidated role in the selective motor neuron pathology in amyotrophic lateral sclerosis (ALS). Among other responsibilities, astrocytes provide important neuronal homeostatic support, however this function is highly compromised in ALS. The establishment of fully human coculture systems can be used to further study the underlying mechanisms of the dysfunctional intercellular interplay, and has the potential to provide a platform for revealing novel therapeutic entry points.MethodsIn this study, we characterised human induced pluripotent stem cell (hiPSC)-derived astrocytes from FUS-ALS patients, and incorporated these cells into a human motor unit microfluidics model to investigate the astrocytic effect on hiPSC-derived motor neuron network and functional neuromuscular junctions (NMJs) using immunocytochemistry and live-cell recordings. FUS-ALS cocultures were systematically compared to their CRISPR-Cas9 gene-edited isogenic control systems.ResultsWe observed a dysregulation of astrocyte homeostasis, which resulted in a FUS-ALS-mediated increase in reactivity and secretion of inflammatory cytokines. Upon coculture with motor neurons and myotubes, we detected a cytotoxic effect on motor neuron-neurite outgrowth, NMJ formation and functionality, which was improved or fully rescued by isogenic control astrocytes. We demonstrate that ALS astrocytes have both a gain-of-toxicity and loss-of-support function involving the WNT/β-catenin pathway, ultimately contributing to the disruption of motor neuron homeostasis, intercellular networks and NMJs.ConclusionsOur findings shine light on a complex, yet highly important role of astrocytes in ALS, and provides further insight in to their pathological mechanisms.Graphical

Effects of short-term unloading and active recovery on human motor unit properties, neuromuscular junction transmission and transcriptomic profile.

Electrophysiological alterations of the neuromuscular junction (NMJ) and motor unit potential (MUP) with unloading are poorly studied. We aimed to investigate these aspects and the underlying molecular mechanisms with short-term unloading and active recovery (AR). Eleven healthy males underwent a 10-day unilateral lower limb suspension (ULLS) period, followed by 21-day AR based on resistance exercise. Quadriceps femoris (QF) cross-sectional area (CSA) and isometric maximum voluntary contraction (MVC) were evaluated. Intramuscular electromyographic recordings were obtained during 10% and 25% MVC isometric contractions from the vastus lateralis (VL). Biomarkers of NMJ molecular instability (serum c-terminal agrin fragment, CAF), axonal damage (neurofilament light chain) and denervation status were assessed from blood samples and VL biopsies. NMJ and ion channel transcriptomic profiles were investigated by RNA-sequencing. QF CSA and MVC decreased with ULLS. Increased CAF and altered NMJ transcriptome with unloading suggested the emergence of NMJ molecular instability, which was not associated with impaired NMJ transmission stability. Instead, increased MUP complexity and decreased motor unit firing rates were found after ULLS. Downregulation of ion channel gene expression was found together with increased neurofilament light chain concentration and partial denervation. The AR period restored most of these neuromuscular alterations. In conclusion, the human NMJ is destabilized at the molecular level but shows functional resilience to a 10-day unloading period at least at relatively low contraction intensities. However, MUP properties are altered by ULLS, possibly due to alterations in ion channel dynamics and initial axonal damage and denervation. These changes are fully reversed by 21days of AR. KEY POINTS: We used integrative electrophysiological and molecular approaches to comprehensively investigate changes in neuromuscular integrity and function after a 10-day unilateral lower limb suspension (ULLS), followed by 21days of active recovery in young healthy men, with a particular focus on neuromuscular junction (NMJ) and motor unit potential (MUP) properties alterations. After 10-day ULLS, we found significant NMJ molecular alterations in the absence of NMJ transmission stability impairment. These findings suggest that the human NMJ is functionally resilient against insults and stresses induced by short-term disuse at least at relatively low contraction intensities, at which low-threshold, slow-type motor units are recruited. Intramuscular electromyography analysis revealed that unloading caused increased MUP complexity and decreased motor unit firing rates, and these alterations could be related to the observed changes in skeletal muscle ion channel pool and initial and partial signs of fibre denervation and axonal damage. The active recovery period restored these neuromuscular changes.

In vivo 3D imaging of human motor units in upper and lower limb muscles

ObjectiveTo assess in-vivo cross-sectional and 3D morphology of human motor units in hand, forearm and lower leg muscles using magnetic resonance imaging (MRI). MethodsDiffusion weighted MRI was used with in-scanner electrical stimulation in healthy controls to image motor units at a single slice in lower leg, forearm and hand muscles (n = 6) and multiple slices in the lower leg for 3D assessment (n = 7). ResultsMotor unit cross-sectional area (CSA) and maximum Feret diameter (FDmax) did not differ between the lower leg (CSA: 22.4 ± 8.4 mm2; FDmax: 8.7 ± 2.4 mm), forearm (CSA: 23.6 ± 14.1 mm2; FDmax: 9.0 ± 3.3 mm) and hand (CSA: 26.8 ± 12.8 mm2 and FDmax: 9.6 ± 2.7 mm) (ANOVA; p = 0.487 and p = 0.587, respectively). Lower leg motor units were 8.0 ± 3.8 cm long with largest CSA in the motor unit’s middle section. 3D motor unit imaging revealed a complex structure with several units splitting and re-forming along their length. ConclusionsMotor unit MRI (MUMRI) can be applied to upper limb muscles, and can reveal the 3D structure of human motor units in-vivo. SignificanceMUMRI provides the first in-vivo 2D images of upper limb motor units and 3D images of lower leg motor units. 3D imaging suggest a more complex human motor unit structure than previously thought.

Effects of reciprocal inhibition and whole-body relaxation on persistent inward currents estimated by two different methods.

Persistent inward currents (PICs) are crucial for initiation, acceleration, and maintenance of motoneuron firing. As PICs are highly sensitive to synaptic inhibition and facilitated by serotonin and noradrenaline, we hypothesised that both reciprocal inhibition (RI) induced by antagonist nerve stimulation and whole‐body relaxation (WBR) would reduce PICs in humans. To test this, we estimated PICs using the well‐established paired motor unit (MU) technique. High‐density surface electromyograms were recorded from gastrocnemius medialis during voluntary, isometric 20‐s ramp, plantarflexor contractions and decomposed into MU discharges to calculate delta frequency (ΔF). Moreover, another technique (VibStim), which evokes involuntary contractions proposed to result from PIC activation, was used. Plantarflexion torque and soleus activity were recorded during 33‐s Achilles tendon vibration and simultaneous 20‐Hz bouts of neuromuscular electrical stimulation (NMES) of triceps surae. ΔF was decreased by RI (n = 15, 5 females) and WBR (n = 15, 7 females). In VibStim, torque during vibration at the end of NMES and sustained post‐vibration torque were reduced by WBR (n = 19, 10 females), while other variables remained unchanged. All VibStim variables remained unaltered in RI (n = 20, 10 females). Analysis of multiple human MUs in this study demonstrates the ability of local, focused inhibition to attenuate the effects of PICs on motoneuron output during voluntary motor control. Moreover, it shows the potential to reduce PICs through non‐pharmacological, neuromodulatory interventions such as WBR. The absence of a consistent effect in VibStim might be explained by a floor effect resulting from low‐magnitude involuntary torque combined with the negative effects of the interventions.Key points Spinal motoneurons transmit signals to skeletal muscles to regulate their contraction. Motoneuron firing partly depends on their intrinsic properties such as the strength of persistent (long‐lasting) inward currents (PICs) that make motoneurons more responsive to excitatory input.In this study, we demonstrate that both reciprocal inhibition onto motoneurons and whole‐body relaxation reduce the contribution of PICs to human motoneuron firing. This was observed through analysis of the firing of single motor units during voluntary contractions.However, an alternative technique that involves tendon vibration and neuromuscular electrical stimulation to evoke involuntary contractions showed less effect. Thus, it remains unclear whether this alternative technique can be used to estimate PICs under all physiological conditions.These results improve our understanding of the mechanisms of PIC depression in human motoneurons. Potentially, non‐pharmacological interventions such as electrical stimulation or relaxation could attenuate unwanted PIC‐induced muscle contractions in conditions characterised by motoneuron hyperexcitability.

Endurance-exercise training adaptations in spinal motoneurones: potential functional relevance to locomotor output and assessment in humans.

It is clear from non-human animal work that spinal motoneurones undergo endurance training (chronic) and locomotor (acute) related changes in their electrical properties and thus their ability to fire action potentials in response to synaptic input. The functional implications of these changes, however, are speculative. In humans, data suggests that similar chronic and acute changes in motoneurone excitability may occur, though the work is limited due to technical constraints. To examine the potential influence of chronic changes in human motoneurone excitability on the acute changes that occur during locomotor output, we must develop more sophisticated recording techniques or adapt our current methods. In this review, we briefly discuss chronic and acute changes in motoneurone excitability arising from non-human and human work. We then discuss the potential interaction effects of chronic and acute changes in motoneurone excitability and the potential impact on locomotor output. Finally, we discuss the use of high-density surface electromyogram recordings to examine human motor unit firing patterns and thus, indirectly, motoneurone excitability. The assessment of single motor units from high-density recording is mainly limited to tonic motor outputs and minimally dynamic motor output such as postural sway. Adapting this technology for use during locomotor outputs would allow us to gain a better understanding of the potential functional implications of endurance training-induced changes in human motoneurone excitability on motor output.

Human motoneuron firing behavior and single motor unit F-wave

In motor control studies, the F-wave (a recurrent discharge evoked by an axonal antidromic volley) widely used for obtaining information on motoneuron pool behavior. However, such F-wave using is a matter of discussion and still has been not validated experimentally. The aim of the present study was investigation of F-wave properties of single firing motor units (MUs) in healthy humans, the properties, which could give evidence for F-wave origin in motoneuron soma and, therefore, could be used for estimation of a relation between MU firing and motoneuron firing behavior. In total, 91 MUs in five muscles of six healthy subjects, during gentle voluntary contractions, were studied. Peri-stimulus time histograms of single MUs were plotted. None of them revealed statistically significant increasing in MU firing probability at the F-wave latency. Analysis of relationships between characteristics of motoneuron firing behavior (mean firing frequency and target interspike interval duration) and properties of F-waves showed their independence. At the same time, it was found that F-waves were recorded in MUs, whose axons possessed the marked supernormal period in excitability recovery cycle after a discharge. Thus, the present results are in contrast to that which should be expected if the F-wave originated in the motoneuronal soma and could provide evidence for motoneuron firing behavior.

Electrical cross-sectional imaging of human motor units in vivo

ObjectivesIn many neuromuscular diseases, weakness results from a disruption in muscle fibres' arrangement within a motor unit. Limitations in current techniques mean that the spatial distribution of fibres in human motor units remains unknown. MethodsA flexible multi-channel electrode was developed and bonded to a clinical electromyography (EMG) needle. Muscle fibre action potentials were localised using a novel deconvolution method. This was tested using simulated data, and in recordings collected from the tibialis anterior muscle of healthy subjects. ResultsSimulated data indicated good localisation reliability across all sections of the electrode except the end sections. A corrected fibre density was estimated up to 1.4 fibres/mm2. Across five recordings from three individuals, between 4 and 14 motor units were detected. Between 1 and 20 muscle fibres were localised per motor unit within the electrode detection area, with up to 220 muscle fibres localised per recording, with overlapping motor unit territories. ConclusionsWe provide the first direct evidence that human motor units spatially overlap, as well as data related to the spatial arrangement of muscle fibres within a motor unit. SignificanceAs well as providing insights into normal human motor physiology, this technology could lead to faster and more accurate diagnosis in patients with neuromuscular diseases.

The Cellular Basis for the Generation of Firing Patterns in Human Motor Units.

Motor units, which comprise a motoneuron and the set of muscle fibers it innervates, are the fundamental neuromuscular transducers for all motor commands. The one to one relationship between a motoneuron and its innervated muscle fibers allow motoneuron firing patterns to be readily measured in humans. In this chapter, we summarize the current understanding of the cellular basis for the generation of firing patterns in human motor units. We provide a brief review of landmark insights from classic studies and then proceed to consider the features of motor unit firing patterns that are most likely to be sensitive estimators of motoneuron inputs and properties. In addition, we discuss recent advances in technology for recording human motor unit firing patterns and highly realistic computer simulations of motoneurons. The final section presents our recent efforts to use the power of supercomputers for implementation of the motoneuron models, with a goal of achieving a true "reverse engineering" approach that maximizes the insights from motor unit firing patterns into the synaptic structure of motor commands.

Electrical Properties of Adult Mammalian Motoneurons.

Motoneurons are the 'final common path' between the central nervous system (that intends, selects, commands, and organises movement) and muscles (that produce the behaviour). Motoneurons are not passive relays, but rather integrate synaptic activity to appropriately tune output (spike trains) and therefore the production of muscle force. In this chapter, we focus on studies of mammalian motoneurons, describing their heterogeneity whilst providing a brief historical account of motoneuron recording techniques. Next, we describe adult motoneurons in terms of their passive, transition, and active (repetitive firing) properties. We then discuss modulation of these properties by somatic (C-boutons) and dendritic (persistent inward currents) mechanisms. Finally, we briefly describe select studies of human motor unit physiology and relate them to findings from animal preparations discussed earlier in the chapter. This interphyletic approach to the study of motoneuron physiology is crucial to progress understanding of how these diverse neurons translate intention into behaviour.

Blockade of 5-HT2 receptors suppresses rate of torque development and motor unit discharge rate during rapid contractions.

Serotonin (5-HT) is a neuromodulator that is critical for regulating the excitability of spinal motoneurons and the generation of muscle torque. However, the role of 5-HT in modulating human motor unit activity during rapid contractions has yet to be assessed. Nine healthy participants (23.7 ± 2.2 yr) ingested 8 mg of the competitive 5-HT2 antagonist cyproheptadine in a double-blinded, placebo-controlled, repeated-measures experiment. Rapid dorsiflexion contractions were performed at 30%, 50%, and 70% of maximal voluntary contraction (MVC), where motor unit activity was assessed by high-density surface electromyographic decomposition. A second protocol was performed where a sustained, fatigue-inducing dorsiflexion contraction was completed before undertaking the same 30%, 50%, and 70% MVC rapid contractions and motor unit analysis. Motor unit discharge rate (P < 0.001) and rate of torque development (RTD; P = 0.019) for the unfatigued muscle were both significantly lower for the cyproheptadine condition. Following the fatigue inducing contraction, cyproheptadine reduced motor unit discharge rate (P < 0.001) and RTD (P = 0.024), whereas the effects of cyproheptadine on motor unit discharge rate and RTD increased with increasing contraction intensity. Overall, these results support the viewpoint that serotonergic effects in the central nervous system occur fast enough to regulate motor unit discharge rate during rapid powerful contractions.NEW & NOTEWORTHY We have shown that serotonin activity in the central nervous system plays a role in regulating human motor unit discharge rate during rapid contractions. Our findings support the viewpoint that serotonergic effects in the central nervous system are fast and are most prominent during contractions that are characterized by high motor unit discharge rates and large amounts of torque development.