Concurrent Achilles tendon vibration and tibial nerve stimulation to estimate persistent inward current strength in motoneurons

Vibratory (Tvib) and sustained (Tsust) torque responses to concurrent Achilles tendon vibration and neuromuscular electrical stimulation applied over the muscle belly (vib+stim) are used as indicators of motoneuron facilitation and, theoretically, persistent inward current strength. However, neuromuscular electrical stimulation (NMES) applied to the nerve trunk may potentiate motoneuronal excitability more than muscle belly NMES, yet it remains unclear whether NMES applied over the nerve evokes robust Tvib and Tsust responses when used during the vib+stim protocol. This study tested whether a nerve-targeted vib+stim protocol elicits Tvib and Tsust responses in the ankle plantar flexors with acceptable intra- and inter-session reliability. Fifteen men performed the vib+stim protocol with NMES applied over the tibial nerve three times across two sessions; twice in a single session (5-min apart) to test intrasession reliability and then again after 48 h to test intersession reliability. Intraclass correlation coefficients (ICC3,1), within-participant coefficients of variation (CV) and pairwise comparisons were used to verify relative and absolute reliability as well as systematic bias. Thirteen men presented Tvib and Tsust responses (response rate of 87%). Intrasession Tvib and Tsust ICCs were >0.73 but inter-session ICCs were <0.5. Although no systematic bias was detected (p>0.05), both intra- and inter-session CVs were large (>10%) for Tvib and Tsust. The Vib+stim protocol with NMES applied over the nerve evoked Tvib and Tsust in almost all participants, but presented a large intra- and inter-session variability. The method does not appear to be effective for assessing motoneuron facilitation in the plantar flexors.

The mechanism underlying transient weakness in myotonia congenita

In addition to the hallmark muscle stiffness, patients with recessive myotonia congenita (Becker disease) experience debilitating bouts of transient weakness that remain poorly understood despite years of study. We performed intracellular recordings from muscle of both genetic and pharmacologic mouse models of Becker disease to identify the mechanism underlying transient weakness. Our recordings reveal transient depolarizations (plateau potentials) of the membrane potential to −25 to −35 mV in the genetic and pharmacologic models of Becker disease. Both Na+ and Ca2+ currents contribute to plateau potentials. Na+ persistent inward current (NaPIC) through NaV1.4 channels is the key trigger of plateau potentials and current through CaV1.1 Ca2+ channels contributes to the duration of the plateau. Inhibiting NaPIC with ranolazine prevents the development of plateau potentials and eliminates transient weakness in vivo. These data suggest that targeting NaPIC may be an effective treatment to prevent transient weakness in myotonia congenita.

The mechanism underlying transient weakness in myotonia congenita

In addition to the hallmark muscle stiffness, patients with recessive myotonia congenita (Becker disease) experience debilitating bouts of transient weakness that remain poorly understood despite years of study. We made intracellular recordings from muscle of both genetic and pharmacologic mouse models of Becker disease to identify the mechanism underlying transient weakness. Our recordings reveal transient depolarizations (plateau potentials) of the membrane potential to −25 to −35 mV in the genetic and pharmacologic models of Becker disease. Both Na+ and Ca2+ currents contribute to plateau potentials. Na+ persistent inward current (NaPIC) through Naγ1.4 channels is the key trigger of plateau potentials and current through Cav1.1 Ca2+ channels contributes to the duration of the plateau. Inhibiting NaPIC with ranolazine prevents the development of plateau potentials and eliminates transient weakness in vivo. These data suggest that targeting NaPIC may be an effective treatment to prevent transient weakness in myotonia congenita.Impact StatementTransient weakness in myotonia congenita is caused by depolarization secondary to activation of persistent Na+ current in skeletal muscle.

An interaction between the III-IV linker and CTD in NaV1.5 confers regulation of inactivation by CaM and FHF

Voltage gated sodium channel (VGSC) activation drives the action potential upstroke in cardiac myocytes, skeletal muscles, and neurons. After opening, VGSCs rapidly enter a non-conducting, inactivated state. Impaired inactivation causes persistent inward current and underlies cardiac arrhythmias. VGSC auxiliary proteins calmodulin (CaM) and fibroblast growth factor homologous factors (FHFs) bind to the channel’s C-terminal domain (CTD) and limit pathogenic persistent currents. The structural details and mechanisms mediating these effects are not clear. Building on recently published cryo-EM structures, we show that CaM and FHF limit persistent currents in the cardiac NaV1.5 VGSC by stabilizing an interaction between the channel’s CTD and III-IV linker region. Perturbation of this intramolecular interaction increases persistent current and shifts the voltage dependence of steady-state inactivation. Interestingly, the NaV1.5 residues involved in the interaction are sites mutated in the arrhythmogenic long QT3 syndrome (LQT3). Along with electrophysiological investigations of this interaction, we present structural models that suggest how CaM and FHF stabilize the interaction and thereby limit the persistent current. The critical residues at the interaction site are conserved among VGSC isoforms, and subtle substitutions provide an explanation for differences in inactivation among the isoforms.

Estimates of persistent inward current in human motor neurons during postural sway

Persistent inward current (PIC) plays a critical role in setting the gain of spinal motor neurons. In humans, most estimates of PIC are made from plantarflexor or dorsiflexor motor units in a seated position. This seated and static posture negates the task-dependent nature of the monoaminergic drive and afferent inhibition that modulate PIC activation. Our purpose was to estimate PIC during both the conventional seated posture and in a more functionally relevant anterior postural sway. We hypothesized that paired motor unit estimates of PIC would be greater when during standing compared with sitting. Soleus motor neuron PIC was estimated via the paired motor unit (PMU) technique. For each motor unit pair, difference in reference unit firing frequency (ΔF) estimates of PIC were made during isometric ramps in plantarflexion force during sitting (conventional approach) and during standing anterior postural sway (new approach). Baseline reciprocal inhibition (RI) was also measured in each posture using the poststimulus time histogram technique. ΔF estimates during standing postural sway were not different [2.64 ± 0.95 pulses/s (pps), P = 0.098] from seated PIC estimates (3.15 ± 1.45 pps) measured from the same motor unit pair. Similarly, reciprocal inhibition at the onset of each task was the same in standing (−0.60 ± 0.32, P = 0.301) and seated (−0.86 ± 0.82) postures. PMU recordings made during standing postural sway met all assumptions that underlay the PMU technique, including rate modulation ≥0.5 pps (3.11 ± 1.90 pps), rate-rate correlation r ≥ 0.7 (0.84 ± 0.13), and time between reference and test unit recruitment ≥1 s (1.83 ± 0.81 s). This study presents a novel, functionally relevant standing method for investigating PIC in humans. NEW & NOTEWORTHY Paired motor unit (PMU) estimates of persistent inward current (PIC) in human soleus motor units are typically made in seated posture. Our study demonstrates that these estimates can be made during standing forward sway, a task that more accurately reflects the postural role of human soleus muscle. PMU recordings made during standing postural sway were validated using all previously published criteria used to test the assumptions of the PMU technique. Standing estimates of PIC did not differ from seated estimates made from the same motor unit pairs.

Impact of the localization of dendritic calcium persistent inward current on the input-output properties of spinal motoneuron pool: a computational study

The goal of this study is to investigate how the dendritic Ca-PIC location influences nonlinear input-output properties and depends on the type of motoneurons across the motoneuron pool. A model motoneuron pool consisting of 10 motoneurons was constructed using a recently developed two-compartment modeling approach that reflected key cell type-associated properties experimentally identified. The dendritic excitability and firing output depended systematically on both the PIC location and the motoneuron type. The PIC onset and offset in the current-voltage ( I–V) relationship tended to occur at more hyperpolarized voltages as the path length to the PIC channels from the soma increased and as the cell type shifted from high- to low-threshold motoneurons. At the same time, the firing acceleration and frequency hysteresis in the frequency-current ( F–I) relationship became faster and larger, respectively. However, the PIC onset-offset hysteresis increased as the path length and the recruitment threshold increased. Furthermore, the gain of frequency-current function before full PIC activation was larger for PIC channels located over distal dendritic regions in low- compared with high-threshold motoneurons. When compared with previously published experimental observations, the modeling concurred when Ca-PIC channels were placed closer to the soma in high- than low-threshold motoneurons in the model motoneuron pool. All of these results suggest that the negative relationship of Ca-PIC location and cell recruitment threshold may underlie the systematic variation in I–V and F–I transformation across the motoneuron pool. NEW & NOTEWORTHY How does the dendritic location of calcium persistent inward current (Ca-PIC) influence dendritic excitability and firing behavior across the spinal motoneuron pool? This issue was investigated developing a model motoneuron pool that reflected key motoneuron type-specific properties experimentally identified. The simulation results point out the negative relationship between the distance of Ca-PIC source from the soma and cell recruitment threshold as a basis underlying the systematic variation in input-output properties of motoneurons over the motoneuron pool.

An evaluation of paired motor unit estimates of persistent inward current in human motoneurons

Persistent inward current (PIC) plays an important role in setting the input-output gain of motoneurons. In humans, these currents are estimated by calculating the difference between synaptic input at motor unit recruitment and derecruitment (ΔF) derived from paired motor unit recordings. The primary objective of this study was to use the relationship between reciprocal inhibition (RI) and PIC to estimate the contribution of PIC relative to other motoneuron properties that result in nonlinear motor unit firing behavior. This study also assessed the contribution of other intrinsic properties (spike threshold accommodation and spike frequency adaptation) to ΔF estimates of PIC in human motor units by using ramps with varying rates of rise and duration. It was hypothesized that slower rates of ramp rise and longer ramp durations would inflate ΔF estimates of PIC, and RI and PIC values would only be correlated during the ramp with the fastest rate of rise and shortest duration when spike threshold accommodation and spike frequency adaptation is minimized. Fourteen university-aged participants took part in this study. Paired motor unit recordings were made from the right soleus muscle during ramp contractions of plantar flexors with three different rates of rise and durations. ΔF estimates of PIC increased with decreased rates of ramp rise ( P < 0.01) and increased ramp durations ( P < 0.01), most likely due to spike frequency adaptation. A correlation ( r = 0.41; P < 0.03) between ΔF and RI provides evidence that PIC is the primary contributor to ΔF in shorter ramps with faster rates of rise.

Cx46 hemichannels contribute to the sodium leak conductance in lens fiber cells

The lens is proposed to have an internal microcirculation system consisting of continuously circulating ionic fluxes that play an essential role in maintaining lens transparency. One of the key components of this system is the sodium leak conductance. Here we investigate the contribution of Cx46 hemichannels to the basal membrane permeability of peripheral fiber cells isolated from transgenic mouse lenses lacking Cx50 or both Cx50 and Cx46 (dKO) using the whole cell patch-clamp technique. Our results show that Cx46 hemichannels were largely closed at a resting voltage of −60 mV in the presence of millimolar divalent cation concentrations. However, even though the vast majority of these channels were closed at −60 mV, a small, persistent, inward current could still be detected. This current could be mostly blocked by exposure to 1 mM La3+ and was not observed in fiber cells isolated from dKO mouse lenses suggesting that it was due to Cx46 hemichannels. In addition, Cx50−/− fiber cells showed increased open channel noise and a depolarized resting potential compared with dKO fiber cells. Exposure of Cx50−/− fiber cells to La3+ hyperpolarized the resting potential to −58 mV, which is similar to the value of resting potential measured in dKO fiber and significantly reduced the open channel noise. In conclusion, these results suggest that Cx46 hemichannels may contribute to the sodium leak conductance in lens fiber cells.

NMDA induces persistent inward and outward currents that cause rhythmic bursting in adult rodent motoneurons

N-methyl-d-aspartate (NMDA) receptors are of critical importance for locomotion in the developing neonatal spinal cord in rats and mice. However, due to profound changes in the expression of NMDA receptors in development between the neonatal stages and adulthood, it is unclear whether NMDA receptors are still an important component of locomotion in the adult rodent spinal cord. To shed light on this issue, we have taken advantage of recently developed preparations allowing the intracellular recording of adult motoneurons that control the tail in the sacrocaudal spinal cord of adult mice and rats. We show that in the adult sacrocaudal spinal cord, NMDA induces rhythmic activity recorded on the ventral roots, often coordinated from left to right, as in swimming motions with the tail (fictive locomotion). The adult motoneurons themselves are intrinsically sensitive to NMDA application. That is, when motoneurons are synaptically isolated with TTX, NMDA still causes spontaneous bursts of rhythmic activity, depending on the membrane potential. We show that these bursts in motoneurons depend on an NMDA-mediated persistent inward current and are terminated by the progressive activation of a persistent outward current. These results indicate that motoneurons, along with the central pattern generator, can actively participate in the production of swimminglike locomotor activity in adult rodents.