Diminished GAD67 Terminals and Ambient GABA Inhibition Associated with Reduced Motor Neuron Recruitment Threshold in the SOD1G93A ALS mouse

Pre-symptomatic studies in mouse models of the neurodegenerative motor neuron (MN) disease, Amyotrophic Lateral Sclerosis (ALS) highlight early alterations in intrinsic and synaptic excitability and have supported an excitotoxic theory of MN death. However, a role for synaptic inhibition in disease development is not sufficiently explored among other mechanisms. Since inhibition plays a role in both regulating motor output and in neuroprotection, we examined the age-dependent anatomical changes in inhibitory presynaptic terminals on MN cell bodies using fluorescent immunohistochemistry for GAD67 (GABA) and GlyT2 (glycine) presynaptic proteins comparing ALS-vulnerable trigeminal jaw closer (JC) motor pools with the ALS-resistant extraocular (EO) MNs in the SOD1G93A mouse model for ALS. Our results indicate differential patterns of temporal changes of these terminals in vulnerable versus resilient MNs and relative differences between SOD1G93A and wild-type (WT) MNs. Notably, we found pre-symptomatic up-regulation in inhibitory terminals in the EO MNs while the vulnerable JC MNs mostly showed a decrease in inhibitory terminals. Specifically, there was a statistically significant decrease in the GAD67 somatic abuttal in the SOD1G93A JC MNs compared to WT around P12. Using in vitro patch-clamp electrophysiology, we found a parallel decrease in the ambient GABA-dependent tonic inhibition in the SOD1G93A JC MNs. While it is unclear if the two mechanisms are directly related, pharmacological blockade of specific subtype of GABAA-a5 receptors suggests that tonic inhibition can control MN recruitment threshold. Furthermore, reduction in tonic GABA current as observed here in the mutant, identifies a putative molecular mechanism explaining our observations of hyperexcitable shifts in JC MN recruitment threshold in the SOD1G93A mouse. Lastly, we showcase non-parametric resampling-based bootstrap statistics for data analyses, and provide the Python code on GitHub for wider reuse.

Pain-induced changes in motor unit discharge depend on recruitment threshold and contraction speed

At high forces, the discharge rates of lower and higher threshold motor units (MU) are influenced in a different way by muscle pain. These differential effects may be particularly important for performing contractions at different speeds since the proportion of lower and higher threshold MUs recruited varies with contraction velocity. We investigated whether MU discharge and recruitment strategies are differentially affected by pain depending on their recruitment threshold (RT), across a range of contraction speeds. Participants performed ankle dorsiflexion sinusoidal-isometric contractions at two frequencies (0.25Hz and 1Hz) and two modulation amplitudes [5% and 10% of the maximum voluntary contraction (MVC)] with a mean target torque of 20%MVC. High-density surface electromyography recordings from the tibialis anterior muscle were decomposed and the same MUs were tracked across painful (hypertonic saline injection) and non-painful conditions. Torque variability, mean discharge rate (MDR), DR variability (DRvar), RT and the delay between the cumulative spike train and the resultant torque output (neuromechanical delay, NMD) were assessed. The average RT was greater at faster contraction velocities (p=0.01) but was not affected by pain. At the fastest contraction speed, torque variability and DRvar were reduced (p<0.05) and MDR was maintained. Conversely, MDR decreased and DRvar and NMD increased significantly during pain at slow contraction speeds (p<0.05). These results show that reductions in contraction amplitude and increased recruitment of higher threshold MUs at fast contraction speeds appears to compensate for the inhibitory effect of nociceptive inputs on lower threshold MUs, allowing the exertion of fast submaximal contractions during pain.

An examination of a potential organized motor unit firing rate and recruitment scheme of an antagonist muscle during isometric contractions

The primary purpose of the present study is to determine if an organized control scheme exists for the antagonist muscle during steady isometric torque. A secondary focus is to better understand how firing rates of the antagonist muscle changes from a moderate- to higher-contraction intensity. Fourteen subjects performed two submaximal isometric trapezoid muscle actions of the forearm flexors that included a linearly increasing, steady force at both 40% and 70% maximum voluntary contraction, and linearly decreasing segments. Surface electromyographic signals of the biceps and triceps brachii were collected and decomposed into constituent motor unit action potential trains. Motor unit firing rate vs. recruitment threshold, motor unit action potential amplitude vs. recruitment threshold, and motor unit firing rate vs. action potential amplitude relationships of the biceps brachii (agonist) and triceps brachii (antagonist) muscles were analyzed. Moderate- to-strong relationships (|r| ³ 0.69) were present for the agonist and antagonist muscles for each relationship with no differences between muscles (p = 0.716, 0.428, 0.182). The y-intercepts of the motor unit firing rate vs. recruitment threshold relationship of the antagonist did not increase from 40% to 70% maximal voluntary contractions (p = 0.96), unlike for the agonist (p = 0.009). The antagonist muscle exhibits a similar motor unit control scheme to the agonist. Unlike the agonist, however, the firing rates of the antagonist did not increase with increasing intensity. Future research should investigate how antagonist firing rates adapt to resistance training and changes in antagonist firing rates in the absence of peripheral feedback.

Characteristics of the Electrophysiological Properties of Neuromuscular Motor Units and Its Adaptive Strategy Response in Lower Extremity Muscles for Seniors with Pre-Sarcopenia: A Preliminary Study

Older adults with sarcopenia, which is an aging-related phenomenon of muscle mass loss, usually suffer from decreases in both strength and functional performance. However, the causality between function loss and physiological changes is unclear. This study aimed to explore the motor unit characteristics of the neurological factors between normal subjects and those with sarcopenia. Five risk-sarcopenia (age: 66.20 ± 4.44), five healthy (age: 69.00 ± 2.35), and twelve young (age: 21.33 ± 1.15) participants were selected. Each participant performed knee extension exercises at a 50% level of maximal voluntary isometric contraction. Next, electromyogram (EMG) signals were collected, and information on each parameter—e.g., motor unit number, recruitment threshold, the slope of the mean firing rate to recruitment threshold, y-intercept, firing rate per unit force, and mean motor unit firing rate (MFR)—was extracted to analyze muscle fiber discrimination (MFD). Meanwhile, force variance was used to observe the stability between two muscle groups. The results suggested that there was no difference between the three groups for motor unit number, recruitment threshold, y-intercept, mean firing rate, and motor unit discrimination (p > 0.05). However, the slope of MFR and firing rate per unit force in the risk-sarcopenia group were significantly higher than in the young group (p < 0.05). Regarding muscle performance, the force variance in the non-sarcopenia group was significantly higher than the young group (p < 0.05), while the risk-sarcopenia group showed a higher trend than the young group. This study demonstrated some neuromuscular characters between sarcopenia and healthy elderly and young people when performing the same level of leg exercise tasks. This difference may provide some hints for discovering aging-related strength and function loss. Future studies should consider combining the in vivo measurement of muscle fiber type to clarify whether this EMG difference is related to the loss of muscle strength or mass before recruiting symptomatic elderly participants for further investigation.

VASTUS LATERALIS MOTOR UNIT RECRUITMENT THRESHOLDS ARE COMPRESSED TOWARDS LOWER FORCES IN OLDER MEN

Background: Aging results in adaptations which may affect the control of motor units. Objective: We sought to determine if younger and older men recruit motor units at similar force levels. Design: Cross-sectional, between-subjects design. Setting: Controlled laboratory setting. Participants: Twelve younger (age = 25 ± 3 years) and twelve older (age = 75 ± 8 years) men. Measurements: Participants performed isometric contractions of the dominant knee extensors at a force level corresponding to 50% maximal voluntary contraction (MVC). Bipolar surface electromyographic (EMG) signals were detected from the vastus lateralis. A surface EMG signal decomposition algorithm was used to quantify the recruitment threshold of each motor unit, which was defined as the force level corresponding to the first firing. Recruitment thresholds were expressed in both relative (% MVC) and absolute (N) terms. To further understand age-related differences in motor unit control, we examined the mean firing rate versus recruitment threshold relationship at steady force. Results: MVC force was greater in younger men (p = 0.010, d = 1.15). Older men had lower median recruitment thresholds in both absolute (p = 0.005, d = 1.29) and relative (p = 0.001, d = 1.53) terms. The absolute recruitment threshold range was larger for younger men (p = 0.020; d = 1.02), though a smaller difference was noted in relative terms (p = 0.235, d = 0.50). These findings were complimented by a generally flatter slope (p = 0.070; d = 0.78) and lower y-intercept (p = 0.009; d = 1.17) of the mean firing rate versus recruitment threshold relationship in older men. Conclusion: Older men tend to recruit more motor units at lower force levels. We speculate that recruitment threshold compression may be a neural adaptation serving to compensate for lower motor unit firing rates and/or denervation and subsequent re-innervation in aged muscle.

Blood Flow Restriction Alters Motor Unit Behavior During Resistance Exercise

We aimed to determine whether blood flow restriction (BFR) alters the characteristics of individual motor units during low-intensity (LI) exercise. Eight men (26.0±3.8 yrs) performed 5 sets of 15 knee extensions at 20% of one-repetition maximum (with and without BFR). Maximal isometric voluntary contractions (MVC) were performed before and after exercise to quantify force decrement. Submaximal isometric voluntary contractions were additionally performed for 18 s, matching trapezoidal target-force trajectories at 40% pre-MVC. EMG activity was recorded from the vastus lateralis muscle. Then, signals were decomposed to extract motor unit recruitment threshold, firing rates and action potential amplitudes (MUAP). Force decrement was only seen after LI BFR exercise (–20.5%; p<0.05). LI BFR exercise also induced greater decrements in the linear slope coefficient of the regression lines between motor unit recruitment threshold and firing rate (BFR: –165.1±120.4 vs. non-BFR: –44.4±33.1%, p<0.05). Finally, there was a notable shift towards higher values of firing rate and MUAP amplitude post-LI BFR exercise. Taken together, our data indicate that LI BFR exercise increases the activity of motor units with higher MUAP amplitude. They also indicate that motor units with similar MUAP amplitudes become activated at higher firing rates post-LI BFR exercise.