Reductions in Motor Unit Firing are Associated with Clinically Meaningful Leg Extensor Weakness in Older Adults.

Weakness, one of the key characteristics of sarcopenia, is a significant risk factor for functional limitations and disability in older adults. It has long been suspected that reductions in motor unit firing rates (MUFRs) are one of the mechanistic causes of age-related weakness. However, prior work has not investigated the extent to which MUFR is associated with clinically meaningful weakness in older adults. Forty-three community-dwelling older adults (mean: 75.4 ± 7.4 years; 46.5% female) and 24 young adults (mean: 22.0 ± 1.8 years; 58.3% female) performed torque matching tasks at varying submaximal intensities with their non-dominant leg extensors. Decomposed surface electromyographic recordings were used to quantify MUFRs from the vastus lateralis muscle. Computational modeling was subsequently used to independently predict how slowed MUFRs would negatively impact strength in older adults. Bivariate correlations between MUFRs and indices of lean mass, voluntary activation, and physical function/mobility were also assessed in older adults. Weak older adults (n = 14) exhibited an approximate 1.5 and 3 Hz reduction in MUFR relative to non-weak older adults (n = 29) at 50% and 80% MVC, respectively. Older adults also exhibited an approximate 3 Hz reduction in MUFR relative to young adults at 80% MVC only. Our model predicted that a 3 Hz reduction in MUFR results in a strength decrement of 11-26%. Additionally, significant correlations were found between slower MUFRs and poorer neuromuscular quality, voluntary activation, chair rise time performance, and stair climb power (r's = 0.31 to 0.43). These findings provide evidence that slowed MUFRs are mechanistically linked with clinically meaningful leg extensor weakness in older adults.

Bifurcation analysis of motoneuronal excitability mechanisms under normal and ALS conditions.

Introduction: Bifurcation analysis allows the examination of steady-state, non-linear dynamics of neurons and their effects on cell firing, yet its usage in neuroscience is limited to single-compartment models of highly reduced states. This is primarily due to the difficulty in developing high-fidelity neuronal models with 3D anatomy and multiple ion channels in XPPAUT, the primary bifurcation analysis software in neuroscience. Methods: To facilitate bifurcation analysis of high-fidelity neuronal models under normal and disease conditions, we developed a multi-compartment model of a spinal motoneuron (MN) in XPPAUT and verified its firing accuracy against its original experimental data and against an anatomically detailed cell model that incorporates known MN non-linear firing mechanisms. We used the new model in XPPAUT to study the effects of somatic and dendritic ion channels on the MN bifurcation diagram under normal conditions and after amyotrophic lateral sclerosis (ALS) cellular changes. Results: Our results show that somatic small-conductance Ca2+-activated K (SK) channels and dendritic L-type Ca2+ channels have the strongest effects on the bifurcation diagram of MNs under normal conditions. Specifically, somatic SK channels extend the limit cycles and generate a subcritical Hopf bifurcation node in the V-I bifurcation diagram of the MN to replace a supercritical node Hopf node, whereas L-type Ca2+ channels shift the limit cycles to negative currents. In ALS, our results show that dendritic enlargement has opposing effects on MN excitability, has a greater overall impact than somatic enlargement, and dendritic overbranching offsets the dendritic enlargement hyperexcitability effects. Discussion: Together, the new multi-compartment model developed in XPPAUT facilitates studying neuronal excitability in health and disease using bifurcation analysis.

Fast Blue and Cholera Toxin-B Survival Guide for Alpha-Motoneurons Labeling: Less Is Better in Young B6SJL Mice, but More Is Better in Aged C57Bl/J Mice.

Fast Blue (FB) and Cholera Toxin-B (CTB) are two retrograde tracers extensively used to label alpha-motoneurons (α-MNs). The overall goals of the present study were to (1) assess the effectiveness of different FB and CTB protocols in labeling α-MNs, (2) compare the labeling quality of these tracers at standard concentrations reported in the literature (FB 2% and CTB 0.1%) versus lower concentrations to overcome tracer leakage, and (3) determine an optimal protocol for labeling α-MNs in young B6SJL and aged C57Bl/J mice (when axonal transport is disrupted by aging). Hindlimb muscles of young B6SJL and aged C57Bl/J mice were intramuscularly injected with different FB or CTB concentrations and then euthanized at either 3 or 5 days after injection. Measurements were performed to assess labeling quality via seven different parameters. Our results show that tracer protocols of lower concentration and shorter labeling durations were generally better in labeling young α-MNs, whereas tracer protocols of higher tracer concentration and longer labeling durations were generally better in labeling aged α-MNs. A 0.2%, 3-day FB protocol provided optimal labeling of young α-MNs without tracer leakage, whereas a 2%, 5-day FB protocol or 0.1% CTB protocol provided optimal labeling of aged α-MNs. These results inform future studies on the selection of optimal FB and CTB protocols for α-MNs labeling in normal, aging, and neurodegenerative disease conditions.

Non-Invasive Transcutaneous Spinal DC Stimulation as a Neurorehabilitation ALS Therapy in Awake G93A Mice: The First Step to Clinical Translation.

Spinal direct current stimulation (sDCS) modulates motoneuron (MN) excitability beyond the stimulation period, making it a potential neurorehabilitation therapy for amyotrophic lateral sclerosis (ALS), a MN degenerative disease in which MN excitability dysfunction plays a critical and complex role. Recent evidence confirms induced changes in MN excitability via measured MN electrophysiological properties in the SOD1 ALS mouse during and following invasive subcutaneous sDCS (ssDCS). The first aim of our pilot study was to determine the clinical potential of these excitability changes at symptom onset (P90-P105) in ALS via a novel non-invasive transcutaneous sDCS (tsDCS) treatment paradigm on un-anesthetized SOD1-G93A mice. The primary outcomes were motor function and survival. Unfortunately, skin damage avoidance limited the strength of applied stimulation intensity, likewise limiting measurable primary effects. The second aim of this study was to determine which orientation of stimulation (anodal vs cathodal, which are expected to have opposing effects) is beneficial vs harmful in ALS. Despite the lack of measured primary effects, strong trends in survival of the anodal stimulation group, combined with an analysis of survival variance and correlations among symptoms, suggest anodal stimulation is harmful at symptom onset. Therefore, cathodal stimulation may be beneficial at symptom onset if a higher stimulation intensity can be safely achieved via subcutaneously implanted electrodes or alternative methods. Importantly, the many logistical, physical, and stimulation parameters explored in developing this novel non-invasive treatment paradigm on unanesthetized mice provide insight into an appropriate and feasible methodology for future tsDCS study designs and potential clinical translation.

Motoneuron excitability dysfunction in ALS: Pseudo-mystery or authentic conundrum?

In amyotrophic lateral sclerosis (ALS), abnormalities in motoneuronal excitability are seen in early pathogenesis and throughout disease progression. Fully understanding motoneuron excitability dysfunction may lead to more effective treatments. Yet decades of research have not produced consensus on the nature, role or underlying mechanisms of motoneuron excitability dysfunction in ALS. For example, contrary to Ca excitotoxicity theory, predictions of motoneuronal hyper-excitability, normal and hypo-excitability have also been seen at various disease stages and in multiple ALS lines. Accordingly, motoneuron excitability dysfunction in ALS is a disputed topic in the field. Specifically, the form (hyper, hypo or unchanged) and what role excitability dysfunction plays in the disease (pathogenic or downstream of other pathologies; neuroprotective or detrimental) are currently unclear. Although several motoneuron properties that determine cellular excitability change in the disease, some of these changes are pro-excitable, whereas others are anti-excitable, making dynamic fluctuations in overall 'net' excitability highly probable. Because various studies assess excitability via differing methods and at differing disease stages, the conflicting reports in the literature are not surprising. Hence, the overarching process of excitability degradation and motoneuron degeneration is not fully understood. Consequently, the discrepancies on motoneuron excitability dysfunction in the literature represent a substantial barrier to our understanding of the disease. Emerging studies suggest that biological variables, variations in experimental protocols, issues of rigor and sampling/analysis strategies are key factors that may underlie conflicting data in the literature. This review highlights potential confounding factors for researchers to consider and also offers ideas on avoiding pitfalls and improving robustness of data.

Adaptive Neural Decoder for Prosthetic Hand Control.

The overarching goal was to resolve a major barrier to real-life prosthesis usability-the rapid degradation of prosthesis control systems, which require frequent recalibrations. Specifically, we sought to develop and test a motor decoder that provides (1) highly accurate, real-time movement response, and (2) unprecedented adaptability to dynamic changes in the amputee's biological state, thereby supporting long-term integrity of control performance with few recalibrations. To achieve that, an adaptive motor decoder was designed to auto-switch between algorithms in real-time. The decoder detects the initial aggregate motoneuron spiking activity from the motor pool, then engages the optimal parameter settings for decoding the motoneuron spiking activity in that particular state. "Clear-box" testing of decoder performance under varied physiological conditions and post-amputation complications was conducted by comparing the movement output of a simulated prosthetic hand as driven by the decoded signal vs. as driven by the actual signal. Pearson's correlation coefficient and Normalized Root Mean Square Error were used to quantify the accuracy of the decoder's output. Our results show that the decoder algorithm extracted the features of the intended movement and drove the simulated prosthetic hand accurately with real-time performance (<10 ms) (Pearson's correlation coefficient >0.98 to >0.99 and Normalized Root Mean Square Error <13-5%). Further, the decoder robustly decoded the spiking activity of multi-speed inputs, inputs generated from reversed motoneuron recruitment, and inputs reflecting substantial biological heterogeneity of motoneuron properties, also in real-time. As the amputee's neuromodulatory state changes throughout the day and the electrical properties and ratio of slower vs. faster motoneurons shift over time post-amputation, the motor decoder presented here adapts to such changes in real-time and is thus expected to greatly enhance and extend the usability of prostheses.

In-silico development and assessment of a Kalman filter motor decoder for prosthetic hand control.

Up to 50% of amputees abandon their prostheses, partly due to rapid degradation of the control systems, which require frequent recalibration. The goal of this study was to develop a Kalman filter-based approach to decoding motoneuron activity to identify movement kinematics and thereby provide stable, long-term, accurate, real-time decoding. The Kalman filter-based decoder was examined via biologically varied datasets generated from a high-fidelity computational model of the spinal motoneuron pool. The estimated movement kinematics controlled a simulated MuJoCo prosthetic hand. This clear-box approach showed successful estimation of hand movements under eight varied physiological conditions with no retraining. The mean correlation coefficient of 0.98 and mean normalized root mean square error of 0.06 over these eight datasets provide proof of concept that this decoder would improve long-term integrity of performance while performing new, untrained movements. Additionally, the decoder operated in real-time (~0.3 ms). Further results include robust performance of the Kalman filter when re-trained to more severe post-amputation limitations in the type and number of motoneurons remaining. An additional analysis shows that the decoder achieves better accuracy when using the firing of individual motoneurons as input, compared to using aggregate pool firing. Moreover, the decoder demonstrated robustness to noise affecting both the trained decoder parameters and the decoded motoneuron activity. These results demonstrate the utility of a proof of concept Kalman filter decoder that can support prosthetics' control systems to maintain accurate and stable real-time movement performance.

Estimating the effects of slicing on the electrophysiological properties of spinal motoneurons under normal and disease conditions.

Although slice recordings from spinal motoneurons (MNs) are being widely used, the effects of slicing on the measured MN electrical properties under normal and disease conditions have not been assessed. Using high-fidelity cell models of neonatal wild-type (WT) and superoxide dismutase-1 (SOD) cells, we examined the effects of slice thickness, soma position within the slice, and slice orientation to estimate the error induced in measured MN electrical properties from spinal slices. Our results show that most MN electrical properties are not adversely affected by slicing, except for cell time constant, cell capacitance, and Ca2+ persistent inward current (PIC), which all exhibited large errors, regardless of the slice condition. Among the examined factors, soma position within the slice appears to be the strongest factor in influencing the magnitude of error in measured MN electrical properties. Transverse slices appear to have the least impact on measured MN electrical properties. Surprisingly, and despite their anatomical enlargement, we found that G85R-SOD MNs experience similar error in their measured electrical properties to those of WT MNs, but their errors are more sensitive to the soma position within the slice than WT MNs. Unless in thick and symmetrical slices, slicing appears to reduce motoneuron type differences. Accordingly, slice studies should attempt to record from MNs at the slice center to avoid large and inconsistent errors in measured cell properties and have valid cell measurements' comparisons. Our results, therefore, offer information that would enhance the rigor of MN electrophysiological data measured from the slice preparation under normal and disease conditions.NEW & NOTEWORTHY Although slice recordings from motoneurons are being widely used, the effects of slicing on the measured motoneuron electrical properties under normal and disease conditions have not been assessed. Using high-fidelity cell models of neonatal WT and SOD cells, we examined the effects of slice thickness, soma position within the slice, and slice orientation. Our results offer information that enhances the rigor of MN electrophysiological data measured from the slice preparation under normal and disease conditions.

A Classification Approach to Recognize the Firing of Spinal Motoneurons in Amyotrophic Lateral Sclerosis.

Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease that affects the nervous system causing muscle weakness, paralysis, leading to death. Given that abnormalities in spinal motoneuron (MN) excitability begin long before symptoms manifest, developing an approach that could recognize fluctuations in MN firing could help in early diagnosis of ALS. This paper introduces a machine learning approach to discriminate between ALS and normal MN firing. The approach is based on two electrophysiological markers; namely, spiking latency and the spike-triggered average signal. The method is examined using data generated from a computational model under systematic variation of MN properties. Such variations mimic the differential dynamic changes in cellular properties that different MN types experience during ALS progression. Our results demonstrate the ability of the approach to accurately recognize ALS firing patterns across the spectrum of examined variations in MN properties.Clinical Relevance- These results represent a proof of concept for using the proposed machine-learning approach in early diagnosis of ALS.

Effects of Neuronic Shutter Observed in the EEG Alpha Rhythm.

The posterior alpha (α) rhythm, seen in human electroencephalogram (EEG), is posited to originate from cycling inhibitory/excitatory states of visual relay cells in the thalamus. These cycling states are thought to lead to oscillating visual sensitivity levels termed the "neuronic shutter effect." If true, perceptual performance should be predictable by observed α phase (of cycling inhibitory/excitatory states) relative to the timeline of afferentiation onto the visual cortex. Here, we tested this hypothesis by presenting contrast changes at near perceptual threshold intensity through closed eyelids to 20 participants (balanced for gender) during times of spontaneous α oscillations. To more accurately and rigorously test the shutter hypothesis than ever before, α rhythm phase and amplitude were calculated relative to each individual's retina-to-primary visual cortex (V1) conduction delay, estimated from the individual's C1 visual-evoked potential (VEP) latency. Our results show that stimulus observation rates (ORs) are greater at a trough than a peak of the posterior α rhythm when phase is measured at the individual's conduction delay relative to stimulus onset. Specifically, the optimal phase for stimulus observation was found to be 272.41°, where ORs are 20.96% greater than the opposing phase of 92.41°. The perception-phase relationship is modulated by α rhythm amplitude and is not observed at lower amplitude oscillations. Collectively, these results provide support to the "neuronic shutter" hypothesis and demonstrate a phase and timing relationship consistent with the theory that cycling excitability in the thalamic relay cells underly posterior α oscillations.

Dendritic distributions of L-type Ca2+ and SKL channels in spinal motoneurons: a simulation study.

Persistent inward currents are important to motoneuron excitability and firing behaviors and also have been implicated in excitotoxicity. In particular, L-type Ca2+ channels, usually located on motoneuron dendrites, play a primary role in amplification of synaptic inputs. However, recent experimental findings on L-type Ca2+ channel behaviors challenge some fundamental assumptions that have been used in interpreting experimental and computational modeling data. Thus, the objectives of this study were to incorporate recent experimental data into an updated, high-fidelity computational model in order to explain apparent inconsistencies and to better elucidate the spatial distributions, expression patterns, and functional roles of L-type Ca2+ and SKL channels. Specifically, the updated model incorporated asymmetric channel activation/deactivation kinetics, depolarization-dependent facilitation, randomness in channel gating, and coactivation of SKL channels. Our simulation results suggest that L-type Ca2+ and SKL channels colocalize primarily on distal dendrites of motoneurons in a punctate expression. Also, punctate expression, as opposed to a homogeneous expression, provides high synaptic current amplification, limits bistability and firing rates, and robustly regulates the Ca2+ persistent inward current, thereby reducing risk of excitotoxicity. The hysteresis and bistability observed experimentally in current-voltage and frequency-current relationships result from the L-type Ca2+ channels' distal location and intrinsic warm-up. Accordingly, our results indicate that punctate expression of L-type Ca2+ and SKL channels is a potent mechanism for regulating excitability, which would provide a strong neuroprotective effect. Our results could provide broader insights into the functional significance of warm-up and punctate expression of ion channels to regulation of cell excitability.NEW & NOTEWORTHY Recent experimental findings on L-type Ca2+ channels challenge fundamental assumptions used in interpreting experimental and computational modeling data. Here, we incorporated recent experimental data into an updated, high-fidelity computational model to explain apparent inconsistencies and better elucidate the distributions, expression patterns, and functional roles of L-type Ca2+ and SKL channels. Our results indicate that punctate expression of L-type Ca2+ and SKL channels is a potent mechanism for regulating motoneuron excitability, providing a strong neuroprotective effect.

Meta-analysis of biological variables' impact on spinal motoneuron electrophysiology data.

Experimental, methodological, and biological variables must be accounted for statistically to maximize accuracy and comparability of published neuroscience data. However, accounting for all variables is nigh impossible. Thus we aimed to identify particularly influential variables within published neurological data, from cat, rat, and mouse studies, via a robust statistical process. Our goal was to develop tools to improve rigor in the collection and analysis of data. We strictly constrained experimental and methodological variables and then assessed four key biological variables within motoneuron research: species, age, sex, and cell type. We quantified intraexperimental and interexperimental variances in 11 commonly reported electrophysiological properties of spinal motoneurons. We first assessed variances without accounting for biological variables and then reassessed them while accounting for all four variables. We next assessed variances with all possible combinations of these four variables. We concluded that some motoneuron properties have low intraexperimental, but high interexperimental, variance; that individual motoneuron properties are impacted differently by biological variables; and that some unexplained variances still remain. We report here the optimal combinations of biological variables to reduce interexperimental variance for all 11 parameters. We also rank each parameter by intra- and interexperimental consistency. We expect these results to assist with design of experimental and analytical methods, and to support accuracy in simulations. Furthermore, although demonstrated on spinal motoneuron electrophysiology literature, our approach is applicable to biological data from all fields of neuroscience. This approach represents an important aid to experimental design, comparison of reported data, and reduction of unexplained variance in neuroscience data.NEW & NOTEWORTHY Our meta-analysis shows the impact of species, age, sex, and cell type on lumbosacral motoneuron electrophysiological properties by thoroughly quantifying variances across literature for the first time. We quantify the variances of 11 motoneuron properties with consideration of biological variables, thus providing specific insights for motoneuron modelers and experimenters, and providing a general methodological template for the quantification of variance in neurological data with the consideration of any experimental, methodological, or biological variables of interest.

The Mechanistic Basis for Successful Spinal Cord Stimulation to Generate Steady Motor Outputs.

Electrical stimulation of the spinal cord is a promising rehabilitation intervention to restore/augment motor function after spinal cord injury (SCI). Combining sensory feedback with stimulation of remaining motor circuits has been shown to be a prerequisite for the functional improvement of SCI patients. However, little is known about the cellular mechanisms potentially underlying this functional benefit in the injured spinal cord. Here, we combine computer simulations with an isolated whole-tissue adult mouse spinal cord preparation to examine synaptic, cellular, and system potentials measured from single motoneurons and ventral roots. The stimulation protocol included separate and combined activation of the sensory inputs (evoked by dorsal root stimulation) and motor inputs (evoked by stimulation of spinal cord tissue) at different frequencies, intensities, and neuromodulatory states. Our data show that, while sensory inputs exhibit short-term depression in response to a train of stimulation, motor inputs exhibit short-term facilitation. However, the concurrent activation of both inputs elicits a stronger and steadier motor output. This effect is enhanced by the application of pharmacological neuromodulators. Furthermore, sensorimotor excitatory postsynaptic potentials (EPSPs) summate sublinearly (i.e., their combination produces an excitatory potential smaller than the sum of the excitatory potentials they would individually produce). However, ventral root compound action potentials (CoAPs) summate supralinearly generating much higher outputs. Computer simulations revealed that the contrasting summation and disproportionality in plasticity between the excitatory postsynaptic potentials (EPSPs) and CoAPs result from the motoneuronal firing threshold acting as an amplitude-selective filter. Together, these results provide the mechanistic basis for the cellular processes contributing to the generation of steady motor outputs using spinal stimulation. This data has great potential to guide the design of more effective stimulation protocols in SCI patients.

The vulnerability of spinal motoneurons and soma size plasticity in a mouse model of amyotrophic lateral sclerosis.

KEY POINTS: Motoneuron soma size is a largely plastic property that is altered during amyotrophic lateral sclerosis (ALS) progression. We report evidence of systematic spinal motoneuron soma size plasticity in mutant SOD1-G93A mice at various disease stages and across sexes, spinal regions and motoneuron types. We show that disease-vulnerable motoneurons exhibit early increased soma sizes. We show via computer simulations that the measured changes in soma size have a profound impact on the excitability of disease-vulnerable motoneurons. This study reveals a novel form of plasticity in ALS and suggests a potential target for altering motoneuron function and survival. ABSTRACT: α-Motoneuron soma size is correlated with the cell's excitability and function, and has been posited as a plastic property that changes during cellular maturation, injury and disease. This study examined whether α-motoneuron somas change in size over disease progression in the G93A mouse model of amyotrophic lateral sclerosis (ALS), a disease characterized by progressive motoneuron death. We used 2D- and 3D-morphometric analysis of motoneuron size and measures of cell density at four key disease stages: neonatal (P10 - with earliest known disease changes); young adult (P30 - presymptomatic with early motoneuron death); symptom onset (P90 - with death of 70-80% of motoneurons); and end-stage (P120+ - with full paralysis of hindlimbs). We additionally examined differences in lumbar vs. sacral vs. cervical motoneurons; in motoneurons from male vs. female mice; and in fast vs. slow motoneurons. We present the first evidence of plastic changes in the soma size of spinal α-motoneurons occurring throughout different stages of ALS with profound effects on motoneuron excitability. Somatic changes are time dependent and are characterized by early-stage enlargement (P10 and P30); no change around symptom onset; and shrinkage at end-stage. A key finding in the study indicates that disease-vulnerable motoneurons exhibit increased soma sizes (P10 and P30). This pattern was confirmed across spinal cord regions, genders and motoneuron types. This extends the theory of motoneuron size-based vulnerability in ALS: not only are larger motoneurons more vulnerable to death in ALS, but are also enlarged further in the disease. Such information is valuable for identifying ALS pathogenesis mechanisms.

Modulation of SK channels regulates locomotor alternating bursting activity in the functionally-mature spinal cord.

The spinal cord contains specialized groups of cells called pattern generators, which are capable of orchestrating rhythmic firing activity in an isolated preparation. Different patterns of activity could be generated in vitro including right-left alternating bursting and bursting in which both sides are synchronized. The cellular and network mechanisms that enable these behaviors are not fully understood. We have recently shown that Ca2+-activated K+ channels (SK channels) control the initiation and amplitude of synchronized bursting in the spinal cord. It is unclear, however, whether SK channels play a similar role in the alternating rhythmic pattern. In the current study, we used a spinal cord preparation from functionally mature mice capable of weight bearing and walking. The present results extend our previous work and show that SK channel inhibition initiates and modulates the amplitude of alternating bursting. We also show that addition of methoxamine, an α1-adrenergic agonist, to a cocktail of serotonin, dopamine, and NMDA evokes robust and consistent alternating bursting throughout the cord.

The effects of model composition design choices on high-fidelity simulations of motoneuron recruitment and firing behaviors.

OBJECTIVE: Computational models often require tradeoffs, such as balancing detail with efficiency; yet optimal balance should incorporate sound design features that do not bias the results of the specific scientific question under investigation. The present study examines how model design choices impact simulation results. APPROACH: We developed a rigorously-validated high-fidelity computational model of the spinal motoneuron pool to study three long-standing model design practices which have yet to be examined for their impact on motoneuron recruitment, firing rate, and force simulations. The practices examined were the use of: (1) generic cell models to simulate different motoneuron types, (2) discrete property ranges for different motoneuron types, and (3) biological homogeneity of cell properties within motoneuron types. MAIN RESULTS: Our results show that each of these practices accentuates conditions of motoneuron recruitment based on the size principle, and minimizes conditions of mixed and reversed recruitment orders, which have been observed in animal and human recordings. Specifically, strict motoneuron orderly size recruitment occurs, but in a compressed range, after which mixed and reverse motoneuron recruitment occurs due to the overlap in electrical properties of different motoneuron types. Additionally, these practices underestimate the motoneuron firing rates and force data simulated by existing models. SIGNIFICANCE: Our results indicate that current modeling practices increase conditions of motoneuron recruitment based on the size principle, and decrease conditions of mixed and reversed recruitment order, which, in turn, impacts the predictions made by existing models on motoneuron recruitment, firing rate, and force. Additionally, mixed and reverse motoneuron recruitment generated higher muscle force than orderly size motoneuron recruitment in these simulations and represents one potential scheme to increase muscle efficiency. The examined model design practices, as well as the present results, are applicable to neuronal modeling throughout the nervous system.

Mixed-mode oscillations in pyramidal neurons under antiepileptic drug conditions.

Subthreshold oscillations in combination with large-amplitude oscillations generate mixed-mode oscillations (MMOs), which mediate various spatial and temporal cognition and memory processes and behavioral motor tasks. Although many studies have shown that canard theory is a reliable method to investigate the properties underlying the MMOs phenomena, the relationship between the results obtained by applying canard theory and conductance-based models of neurons and their electrophysiological mechanisms are still not well understood. The goal of this study was to apply canard theory to the conductance-based model of pyramidal neurons in layer V of the Entorhinal Cortex to investigate the properties of MMOs under antiepileptic drug conditions (i.e., when persistent sodium current is inhibited). We investigated not only the mathematical properties of MMOs in these neurons, but also the electrophysiological mechanisms that shape spike clustering. Our results show that pyramidal neurons can display two types of MMOs and the magnitude of the slow potassium current determines whether MMOs of type I or type II would emerge. Our results also indicate that slow potassium currents with large time constant have significant impact on generating the MMOs, as opposed to fast inward currents. Our results provide complete characterization of the subthreshold activities in MMOs in pyramidal neurons and provide explanation to experimental studies that showed MMOs of type I or type II in pyramidal neurons under antiepileptic drug conditions.

SK channel inhibition mediates the initiation and amplitude modulation of synchronized burst firing in the spinal cord.

Burst firing in motoneurons represents the basis for generating meaningful movements. Neuromodulators and inhibitory receptor blocker cocktails have been used for years to induce burst firing in vitro; however, the ionic mechanisms in the motoneuron membrane that contribute to burst initiation and amplitude modulation are not fully understood. Small conductance Ca2+-activated potassium (SK) channels regulate excitatory inputs and firing output of motoneurons and interneurons and therefore, are a candidate for mediating bursting behavior. The present study examines the role of SK channels in the generation of synchronized bursting using an in vitro spinal cord preparation from adult mice. Our results show that SK channel inhibition is required for both initiation and amplitude modulation of burst firing. Specifically, administration of the synaptic inhibition blockers strychnine and picrotoxin amplified the spinal circuit excitatory drive but not enough to evoke bursting. However, when SK channels were inhibited using various approaches, the excitatory drive was further amplified, and synchronized bursting was always evoked. Furthermore, graded SK channel inhibition modulated the amplitude of the burst in a dose-dependent manner, which was reversed using SK channel activators. Importantly, modulation of neuronal excitability using multiple approaches failed to mimic the effects of SK modulators, suggesting a specific role for SK channel inhibition in generating bursting. Both NMDA (N-methyl-d-aspartate) and AMPA (α-amino-3-hydroxy-5-methylisoxazole-4-propionate) receptors were found to drive the synchronized bursts. The blocking of gap junctions did not disturb the burst synchrony. These results demonstrate a novel mechanistic role for SK channels in initiating and modulating burst firing of spinal motoneurons.NEW & NOTEWORTHY This study demonstrates that cholinergic inhibition or direct blockade of small conductance Ca2+-activated potassium (SK) channels facilitates burst firing in spinal motoneurons. The data provide a novel mechanistic explanation for synchronized bursting initiation and amplitude modulation through SK channel inhibition. Evidence also shows that synchronized bursting is driven by NMDA (N-methyl-d-aspartate) and AMPA (α-amino-3-hydroxy-5-methylisoxazole-4-propionate) receptors and that gap junctions do not mediate motoneuron synchronization in this behavior.

Experimental Design and Data Analysis Issues Contribute to Inconsistent Results of C-Bouton Changes in Amyotrophic Lateral Sclerosis.

The possible presence of pathological changes in cholinergic synaptic inputs [cholinergic boutons (C-boutons)] is a contentious topic within the ALS field. Conflicting data reported on this issue makes it difficult to assess the roles of these synaptic inputs in ALS. Our objective was to determine whether the reported changes are truly statistically and biologically significant and why replication is problematic. This is an urgent question, as C-boutons are an important regulator of spinal motoneuron excitability, and pathological changes in motoneuron excitability are present throughout disease progression. Using male mice of the SOD1-G93A high-expresser transgenic (G93A) mouse model of ALS, we examined C-boutons on spinal motoneurons. We performed histological analysis at high statistical power, which showed no difference in C-bouton size in G93A versus wild-type motoneurons throughout disease progression. In an attempt to examine the underlying reasons for our failure to replicate reported changes, we performed further histological analyses using several variations on experimental design and data analysis that were reported in the ALS literature. This analysis showed that factors related to experimental design, such as grouping unit, sampling strategy, and blinding status, potentially contribute to the discrepancy in published data on C-bouton size changes. Next, we systematically analyzed the impact of study design variability and potential bias on reported results from experimental and preclinical studies of ALS. Strikingly, we found that practices such as blinding and power analysis are not systematically reported in the ALS field. Protocols to standardize experimental design and minimize bias are thus critical to advancing the ALS field.

The transformation of synaptic to system plasticity in motor output from the sacral cord of the adult mouse.

Synaptic plasticity is fundamental in shaping the output of neural networks. The transformation of synaptic plasticity at the cellular level into plasticity at the system level involves multiple factors, including behavior of local networks of interneurons. Here we investigate the synaptic to system transformation for plasticity in motor output in an in vitro preparation of the adult mouse spinal cord. System plasticity was assessed from compound action potentials (APs) in spinal ventral roots, which were generated simultaneously by the axons of many motoneurons (MNs). Synaptic plasticity was assessed from intracellular recordings of MNs. A computer model of the MN pool was used to identify the middle steps in the transformation from synaptic to system behavior. Two input systems that converge on the same MN pool were studied: one sensory and one descending. The two synaptic input systems generated very different motor outputs, with sensory stimulation consistently evoking short-term depression (STD) whereas descending stimulation had bimodal plasticity: STD at low frequencies but short-term facilitation (STF) at high frequencies. Intracellular and pharmacological studies revealed contributions from monosynaptic excitation and stimulus time-locked inhibition but also considerable asynchronous excitation sustained from local network activity. The computer simulations showed that STD in the monosynaptic excitatory input was the primary driver of the system STD in the sensory input whereas network excitation underlies the bimodal plasticity in the descending system. These results provide insight on the roles of plasticity in the monosynaptic and polysynaptic inputs converging on the same MN pool to overall motor plasticity.