The Journal of Physiology

Persistent inward currents (PICs) govern motoneuron output and are influenced by diffuse neuromodulation and local inhibition. When large diameter afferent feedback is lost, as in some neurological conditions, PICs might additionally amplify and prolong synaptic inputs. Here, we examined whether reducing Ia afferent transmission via ischaemic nerve block alters PIC contribution to tibialis anterior (TA) motor unit (MU) discharge. Across two experiments 12 adults (5 female) performed triangular-shaped isometric dorsiflexion to 30% (Experiments 1 and 2) and 50% (Experiment 2) maximum voluntary force (MVF) at baseline, after a 20-minute rest (control), and during occlusion after inducing an ischaemic nerve block, confirmed by abolition of the soleus H-reflex. TA myoelectrical activity measured during contractions was decomposed into MU spike trains, and from smoothed MU discharges, discharge rate hysteresis ({Delta}F) and ascending non-linearity (brace height) were quantified. Results from Experiment 1 involving contractions matched to absolute force levels revealed increased peak discharge rate, {Delta}F, and brace height post-occlusion. However, {Delta}F normalised to maximal theoretical hysteresis did not change across time points. In Experiment 2, where MVF was reassessed at each timepoint and contractions were matched to relative force, peak discharge rate, normalised {Delta}F and brace height increased post-occlusion compared to pre-, across both contraction intensities. {Delta}F only increased post-occlusion at 50% MVF, with no changes at 30% MVF. These results show that ischaemic block of large-diameter axons, likely reducing reciprocal inhibition, increases PIC contribution to discharge rate modulation, highlighting the role of Ia afferent input in shaping motoneuron output in humans.

Motoneuron subtypes exhibit distinct firing properties that are critical for the graded control of muscle force. A key determinant of these differences is the medium afterhyperpolarization (mAHP), which shapes discharge rate and firing gain. While subtype-specific variation in mAHP properties has traditionally been attributed to differences in small-conductance calcium-activated potassium (SK) channel expression, emerging evidence suggests that additional conductances may contribute. Here, we investigated the role of hyperpolarization-activated cyclic nucleotide-gated (HCN) channels in regulating the mAHP and excitability of mouse spinal motoneurons during postnatal development. Using whole-cell patch-clamp recordings, we show that, by the onset of the third postnatal week, an h current (Ih) is active at resting potential in fast motoneurons and is correlated with the amplitude of the mAHP. Pharmacological blockade of HCN channels with ZD7288 increased mAHP amplitude in fast but not slow motoneurons, without affecting mAHP duration, indicating a subtype-specific contribution to mAHP amplitude. In line with the mAHP regulating firing gain, ZD7288 also reduced firing gain in fast but not slow motoneurons. These findings support a contribution of HCN channel activity to the regulation of mAHP amplitude and firing gain in fast motoneurons, highlighting a potential interaction between Ih and SK channel-dependent mechanisms in shaping motoneuron excitability. Key PointsO_LIThe amplitude of the medium afterhyperpolarization (mAHP) is negatively correlated with h-current (Ih) amplitude measured near resting potential in mouse lumbar motoneurons. C_LIO_LIPharmacological blockade of HCN channels selectively increases mAHP amplitude in fast, delayed firing alpha motoneurons, with no effect observed in slow, immediate firing alpha motoneurons. C_LIO_LIInhibition of HCN channels reduces firing gain in fast motoneurons, while slow motoneurons remain unaffected. C_LIO_LIHCN channels regulate firing gain in fast motoneurons, at least in part, through modulation of mAHP amplitude. C_LI

Resistance training lasting a few weeks increases maximal force mainly through neural adaptations that enhance the drive from the nervous system to muscle. While these adaptations have been well documented at the motor unit (MU) level during submaximal force contractions, the mechanisms underlying force increases during maximal voluntary contractions are poorly understood. This is due to a classic technical limitation in tracking MUs longitudinally during maximal force tasks. Here, we solved this technical challenge, enabling the investigation of MU adaptations during MVCs in both the trained and untrained limbs following unilateral resistance training. High-density surface electromyography was recorded from the biceps brachii of both limbs before and after a 4-week unilateral resistance-training intervention, and the same MUs were longitudinally tracked across sessions during MVCs by concatenation of three MVC trials of ~5-s each.Unilateral training increased maximal force in the trained limb (+16%) and induced strength transfer to the untrained limb (+8%). In both limbs, maximal contractions after training were characterized by greater EMG amplitude, faster muscle-fiber conduction velocity, and higher MU discharge rates, indicating enhanced neural drive to the motoneuron pool. These adaptations were strongly associated with improvements in maximal force (R2 > 0.7 for all). Importantly, longitudinal MU tracking revealed a non-uniform adaptation across the MU pool: MUs with higher baseline conduction velocity, indicative of higher recruitment threshold, exhibited the largest pre-post increases in discharge rate, whereas lower-threshold units showed smaller changes. Collectively, these findings demonstrate that gains in maximal force and their transfer to the untrained limb are primarily mediated by enhanced rate coding of higher-threshold MUs during MVCs.

The carotid body (CB) is the key peripheral oxygen sensor. CB mitochondria are hypothesised to be uniquely adapted with unusually low intrinsic oxygen affinity which, in association with nitric oxide (NO) and reactive oxygen species signalling, enables acute responsiveness to hypoxia. However, CB mitochondrial physiology or intrinsic oxygen affinity have never been measured directly. We sought to address this key gap by isolating sheep CB mitochondria and comprehensively characterising their phenotype and contrasting them to a non-oxygen sensing tissue, left ventricular myocardium (LV). High resolution respirometry, liquid chromatography mass spectrometry, enzymatic assays and in silico modelling were used to characterise mitochondrial content, aerobic capacity, oxygen affinity, complex subunit abundance and activity, H2O2 production and NO sensitivity in ovine CB and LV. Mitochondrial oxygen affinity (P50 = 0.089 mmHg) was lower in the CB than the LV (P50 = 0.058 mmHg; p = 0.005). Whilst mitochondrial content was lower in the CB, CB mitochondria had higher respiratory rates and enzymatic activity than LV. H2O2 production and NO sensitivity were similar in the two tissues. While intrinsic mitochondrial oxygen affinity is slightly lower in the oxygen sensing CB than in the non-oxygen sensing LV, this difference is small. Hence, any role of mitochondria in CB oxygen sensing is not due to an intrinsic difference in the O2 affinity of cytochrome oxidase due to differential expression of its subunits. Instead, this work suggests that differences in O2 affinity in vivo are secondary to other factors, perhaps including NO, that alter mitochondrial O2 affinity.

Intense unilateral muscle contractions evoke measurable activity within the contralateral neuroaxis, which can be detected with surface electromyographic activity in the resting homologous muscle. Physiological mirror activity (PMA), the unintentional increase in contralateral muscle excitation, has been implicated in cross-limb interactions and adaptations. Despite longstanding observations of PMA, it remains unknown whether this low-level muscle excitation influences local muscle metabolism. We addressed this question using a vascular occlusion test in 10 healthy adults. Surface electromyography and near-infrared spectroscopy-derived measures of tissue oxygen saturation and muscle oxygen consumption (mVO2) were obtained from the resting left forearm during vascular occlusion at rest and during fatiguing unimanual contractions of the right hand. PMA in the contralateral resting arm was greater during unimanual fatigue than during rest (mean difference: 8.9%AA, 95% CI: 4.1 to 13.8; p = 0.002, g = 1.20). This increase was accompanied by a steeper rate of tissue oxygen desaturation (mean difference: -0.132 %{middle dot}s-1, 95% CI: -0.227 to -0.037; p = 0.012, g = -0.91) and greater mVO2 (mean difference: 0.188 mL O2{middle dot}min-1{middle dot}100 g-1, 95% CI: 0.057 to 0.320; p = 0.010, g = 0.94). Greater PMA was associated with both a faster rate of oxygen desaturation (r = -0.85, 95% CI: -0.96 to -0.46, p = 0.002) and greater mVO2 (r = 0.78, 95% CI: 0.28 to 0.94, p = 0.008). These findings suggest that PMA is accompanied by increased local metabolic demand, consistent with a coupling between unintentional muscle excitation and oxygen extraction in the resting limb.

Descending corticospinal and afferent pathways underlying spinally evoked motor potential both contribute to motor output, yet how their interaction at the spinal and peripheral levels is organized spatially within a muscle remains unclear. This study investigated the spatiotemporal characteristics of descending modulation of spinally evoked motor potentials by combining subthreshold transcranial magnetic stimulation (TMS) with transcutaneous spinal cord stimulation (tSCS). In Experiment 1, spinally evoked motor potentials were recorded from multiple lower-limb muscles at various interstimulus intervals (ISIs) defined relative to central conduction time (CCT). Subthreshold TMS facilitated spinally evoked motor potentials from CCT onward across all recorded muscles, with additional bilateral facilitation observed at longer ISIs. In Experiment 2, high-density surface electromyography (HDsEMG) revealed distinct intramuscular activation patterns in the tibialis anterior. The center of gravity (CoG) of TMS-induced motor evoked potentials was located more proximally than that of spinally evoked motor potentials. Notably, the CoG of facilitation maps was shifted further proximally than that of both single-stimulus responses. These findings suggest that descending and afferent inputs preferentially recruit partially distinct motoneuron pools within the same muscle. The proximal bias of facilitation indicates recruitment of additional motoneurons rather than uniform amplification of existing activity. Together, these results demonstrate that the interaction between descending and afferent inputs is both timing-dependent and spatially non-uniform, providing new insight into sensorimotor integration in the human lower limb.

Mass is a fundamental aspect of muscle contractile function, yet the inertial effects of inactive muscle mass is generally neglected in modeling and not quantified in studies on small muscles or isolated fibers. However, during submaximal contractions, inactive muscle tissue may take longer to be accelerated by active fibers, and may be subject to prolonged deceleration, both of which may potentially reduce force development and work output. We sought to test if inactive tissue mass imposes an inertial penalty on muscle performance, using in situ sinusoidal work-loop experiments on rat plantaris muscles. Regional fascicle dynamics, measured across supramaximal and submaximal levels of activation, showed that decreasing activation significantly reduced fascicle strain and increased both shortening and lengthening latency. Contrary to our predictions, however, reductions in work, beyond those explained by decreased fascicle strain, were negligible. Normalized work did not decline disproportionately relative to force, suggesting no clear inertial penalty on work at this muscle size. Our findings suggest that while inactive muscle mass influences the dynamics of submaximal contractions, its impact on work during submaximal contractions at small muscle sizes is limited.

Sympathetic preganglionic neurons (SPNs) provide the sole spinal output to the peripheral sympathetic nervous system. Although sympathetic control is traditionally attributed to synaptic integration within the spinal cord and ganglia, the reliability of spike propagation along SPN axons themselves has received little attention. Here, and in companion papers, we show that axonal conduction in adult mouse thoracic SPNs is highly modifiable and constitutes a critical site of sympathetic gain control. Using an ex vivo preparation preserving intact paravertebral and splanchnic pathways while blocking synaptic transmission, we recorded compound action potentials evoked across multiple ganglia. Slower-conducting, unmyelinated SPN axons, particularly those with branching axons traversing the interganglionic nerve (IGN), exhibited pronounced, temperature-dependent conduction failures. Elevation of temperature produced membrane hyperpolarization and loss of conduction, consistent with activation of temperature-sensitive K2P leak channels, as supported by pharmacological evidence. Pharmacological activation of TREK-family channels with riluzole or arachidonic acid preferentially suppressed conduction in these axons. In contrast, blockade of voltage-gated K+ channels with 4-aminopyridine (4-AP) robustly facilitated conduction, recruited previously silent axons, and restored propagation under conditions of temperature-induced failure. Surprisingly, tetraethylammonium (TEA) block of K+ channels were without effect or depressant. Transmitter systems further shaped axonal reliability: agonists and antagonists of GABAA receptors, as well as cholinergic manipulations, selectively depressed conduction in slow, branching axons. Together, these findings establish SPN axons, particularly slow-conducting branching fibers, as an active and dynamically regulated substrate for sympathetic output control, revealing a presynaptic mechanism with implications for autonomic physiology and disease. SIGNIFICANCESympathetic output is commonly viewed as being regulated primarily through synaptic integration within spinal and autonomic circuits, while axons are often treated as passive transmission elements. Emerging evidence suggests this assumption is incomplete, particularly in slowly conducting and highly branched sympathetic preganglionic neuron (SPN) axons that may operate near the limits of conduction reliability. This study identifies branch point conduction as a dynamic and pharmacologically modifiable control mechanism governing sympathetic signal transmission. By demonstrating selective vulnerability of distinct SPN populations and revealing strong modulation by potassium channel mechanisms, these findings establish axonal conduction security as an underappreciated site of autonomic gain control. These mechanisms may represent novel therapeutic targets for restoring autonomic function after spinal cord injury and related disorders.

Optogenetic defibrillation uses light-gated ion channels to terminate cardiac arrhythmias through targeted illumination. Previous studies assessed the feasibility of using either cation (e.g. ChR2) or anion (e.g. GtACR1) non-selective channels, both of which depolarise resting cardiomyocytes upon photoactivation. In contrast, recently identified light-gated K+-channels (e.g. WiChR) suppress cardiomyocyte activity while maintaining the membrane potential near its resting state. Here, we use biophysically detailed simulations to compare the defibrillation potential of ChR2, GtACR1, and WiChR. Single-cell simulations show that activation of ChR2 and GtACR1 markedly increase diastolic intracellular Ca2+ concentration (by 42.6% and 52.6%, respectively), whereas WiChR induces only minimal changes (4.0% increase), suggesting a lower pro-arrhythmogenic risk. WiChR activation, however, slightly increases intracellular Na+ levels (by 15.1% compared to 0.1% and 3.4% for ChR2 and GtACR), consistent with the residual Na+ permeability of all currently available K+-selective channelrhodopsins. Simulations of human ventricles and atria demonstrate that GtACR1 most effectively terminates re-entrant arrhythmias at low light intensities, while WiChR achieves comparable efficacy at light levels [≥]5 mW/mm2. Complementary tissue-scale simulations reveal that defibrillation is either based on depolarisation within the excitable gap, followed by fast Na+ channel inactivation (depolarising variants ChR2 and GtACR1), or based on a reduction in membrane resistance supporting arrhythmia termination at sufficiently high light levels (large-conductance ion channels GtACR1 and WiChR). Overall, our findings identify channelrhodopsin ion selectivity as a key determinant of both arrhythmia termination success and mechanisms underlying defibrillation. Key points summaryO_LIWe use computational simulations to compare non-selective cation (ChR2), anion (GtACR1), and K+-selective channelrhodopsins (WiChR) for optogenetic termination of re-entrant arrhythmia. C_LIO_LISingle-cardiomyocyte simulations suggest that ChR2 and GtACR1 activation can cause progressive accumulation of intracellular Ca2+, which is minimised when using WiChR. C_LIO_LISimulations of human left ventricles and atria indicate that GtACR1 is most effective in terminating re-entrant arrhythmia at low light intensities, while WiChR becomes similarly effective at higher intensities. C_LIO_LITissue-scale simulations indicate distinct defibrillation mechanisms: Excitable gap extinction by de-novo action potential initiation followed by inactivation of fast Na+ channels for depolarising channelrhodopsins (ChR2, GtACR1), and reduction in membrane resistance for the large-conductance channels (GtACR1, WiChR), effectively clamping the membrane potential at each channels reversal potential at high light levels. C_LI

Perinatal opioid exposure is a prevalent clinical concern linked to respiratory instability and adverse infant outcomes. The opioid buprenorphine is prescribed as a medication for opioid use disorder during pregnancy and used to treat neonatal opioid withdrawal syndrome, yet its direct effects on neonatal control of breathing have not been examined. Here, we asked how acute buprenorphine exposure affects breathing at rest, and during chemoreceptor stimulation. Using dual-chamber head-out plethysmography, we measured pulmonary ventilation rate ([V]I) and metabolic rate in awake male and female Sprague-Dawley neonatal rats on postnatal days 4-5 (P4-5) during eupnea and a hypoxic-hypercapnic (HH) challenge. The effects of buprenorphine and two opioid receptor antagonists, naloxone hydrochloride, or peripherally restricted naloxone methiodide, were assessed using a repeated measures design. [V]I during eupnea and HH were markedly depressed following buprenorphine administration. Buprenorphine reduced [V]O2 and [V]CO2 and produced ventilatory equivalents for O2 and CO2 consistent with frank hypoventilation, driven by reduced breathing frequency and tidal volume (VT). When administered after buprenorphine, neither naloxone hydrochloride nor naloxone methiodide could rescue the buprenorphine-mediated hypoventilation in eupnea or during HH. In contrast, pre-treatment with either naloxone hydrochloride or naloxone methiodide attenuated buprenorphine-induced hypoventilation by preserving VT. These findings demonstrate that neonatal protective chemoreceptor reflexes are depressed by buprenorphine and suggest that pre-treatment with a peripheral opioid receptor antagonist could mitigate buprenorphine-induced hypoventilation without inducing opioid withdrawal. Key PointsO_LIAcute buprenorphine exposure significantly depressed pulmonary ventilation rate ([V]I) during eupnea and hypoxic hypercapnia (HH) in awake neonatal rats. C_LIO_LIBuprenorphine-induced hypoventilation was driven by reduced tidal volume (VT) and breathing frequency. C_LIO_LIBuprenorphine also reduced oxygen consumption ([V]O2) and carbon dioxide production ([V]CO2). C_LIO_LINaloxone given after buprenorphine failed to reverse hypoventilation. C_LIO_LIIn contrast, pre-treatment with either naloxone hydrochloride or peripherally restricted naloxone methiodide mitigated buprenorphine-induced hypoventilation by preserving VT. C_LI

Individuals with cervical spinal cord injury (SCI) often exhibit asymmetric recovery of upper-limb function, with greater weakness in elbow extensors than flexors. To determine whether muscle-specific changes in motor unit (MU) behavior contribute to this disparity, we identified MU firing instants from high-density surface electromyography to characterize MU firing characteristics in the biceps brachii (BIC) and triceps brachii (TRI) of individuals with cervical SCI (n = 20) and non-injured controls (n = 18). We quantified rate-coding behavior and metrics related to persistent inward currents (PICs), including onset-offset hysteresis ({Delta}F), ascending firing rate nonlinearity, and self-sustained firing. At the group level, BIC MUs in SCI participants showed reduced rate coding and altered ascending firing rate nonlinearity relative to controls. In contrast, TRI MUs showed no clear group-level differences. However, subgroup analysis revealed that SCI participants with low-strength during extension (n = 9) exhibited lower {Delta}F and longer self-sustained firing durations in TRI MUs than those with high-strength (n = 6). In BIC, SCI participants with low-strength during flexion (n = 8) showed reduced rate-coding behavior relative to high-strength SCI participants (n = 9), with no differences in PIC-related metrics. Together, these results demonstrate muscle-specific alterations in MU firing after cervical SCI that may relate to strength recovery or preservation and underscore the need for nuanced analyses in heterogeneous SCI populations. Key pointsO_LIRate coding and nonlinear firing behaviors are significantly altered in the biceps brachii, but not triceps brachii, of participants with cervical spinal cord injury. C_LIO_LIStrength based subgroup analyses revealed muscle-specific differences in motor unit behaviors that may be associated with strength preservation or recovery following spinal cord injury. C_LIO_LIFunctional heterogeneity following spinal cord injury may mask group differences in motor unit behaviors and warrants careful interpretation of results of future studies. C_LI

Regular exercise induces adaptations in skeletal muscle and other organ systems to improve physical performance and overall health. Exercise results in phosphorylation of 5 AMP-activated protein kinase (AMPK) at threonine 172 (T172) of the 2 subunit; however, the role of this activation in cellular and functional adaptations has not been elucidated. To this end, we subjected non-activatable Ampk2(T172A) knock-in (KI) adult mice and wild-type (WT) littermates to 4 weeks of voluntary wheel running (VWR). Exercise training led to significant improvements in endurance capacity, maximal oxygen consumption ([Formula]O2max), and glucose tolerance, as well as skeletal muscle IIb-to-IIa fiber type shift in both WT and KI mice. Contrastingly, VWR resulted in increased mitochondrial OxPhos protein expression, mitochondrial volume density, and capillary density in skeletal muscle of WT but not KI mice. Exercise-induced improvements of mitochondrial respiration and conductance revealed by high-resolution respirometry of isolated mitochondria were blunted in KI mice. Therefore, for the first time, we reveal that AMPK2 T172 activation is required for exercise training-induced mitochondrial biogenesis, improvement of mitochondrial respiratory function, and angiogenesis in skeletal muscle, but that these adaptations are not solely responsible for improved [Formula]O2max and exercise endurance capacity. Significance StatementExercise is the most effective lifestyle intervention for promoting health and preventing chronic diseases through adaptive changes in skeletal muscle and many other tissues/organs. AMPK is an energy sensor and signaling regulator for exercise-induced skeletal muscle adaptation, yet its functional role and the impact on exercise capacity have been studied in mouse genetic models wherein protein stoichiometry is disrupted. Using non-activatable Ampk2(T172A) knock-in mice, we ascertained that AMPK2 activation via T172 phosphorylation is required for endurance training-induced mitochondrial and angiogenic adaptations in skeletal muscle. Importantly, these adaptations are not required for improved exercise capacity, challenging the prevailing concept that increased mitochondrial content and function and microvasculature are the sole driving factors for the performance gains with endurance training.

The pelvic floor muscles (PFM) are critical for maintaining continence and are a primary target of physiotherapy training to manage urinary incontinence. PFM training relies on voluntarily activating this muscle group, limiting its translation to neurological populations where recovery of bladder function is a priority. Indirect evidence suggests that sensory feedback from the lower limb can modulate PFM activity, which may provide alternative strategies for PFM training. Cutaneous reflexes have been used as a proxy to study how sensory inputs from the skin influence motoneuron excitability. To explore the feasibility of eliciting cutaneous reflexes in the PFM and their role in controlling PFM activity, this study examined: 1) the input-output relationship and 2) the nerve-specificity of PFM cutaneous reflex responses from tibial and superficial peroneal nerve stimulation. Twenty-one neurologically intact adults participated in this study. We recorded PFM and lower leg muscle electromyography while participants received cutaneous stimulation to the right distal tibial nerve, bilateral distal tibial nerve, or right superficial peroneal nerve in a standing position. We delivered stimulation at the intensity below motor threshold (MT), 1.2 x MT and 1.5 x MT and quantified tibial-PFM reflex amplitude over a 50-150 ms window after stimulation. PFM reflex responses were evoked from both nerves stimulation. Reflex amplitude increased with stimulus intensity with tibial nerve stimulation but not with superficial peroneal nerve stimulation. Bilateral tibial nerve stimulation evoked larger responses compared to unilateral stimulation. These findings support the existence of neural connections between lower limb afferents and the PFM, and open up possibilities for designing rehabilitation strategies to manage pelvic health conditions in people with neurological disorders. New & NoteworthyO_LICutaneous sensory feedback from the foot, specifically that related to limb loading, can evoke reflex responses in the pelvic floor muscles C_LIO_LINerve-specific modulation was observed. Reflex amplitudes in the pelvic floor muscles increased with tibial nerve stimulation intensity but not with superficial peroneal nerve stimulation. C_LI

Background The relative contributions of molecular size, electrostatic charge, and filtration rate to glomerular transport remain controversial. We hypothesized that glomerular sieving data contain a limited number of underlying transport modes that can be identified directly from experimental measurements. Methods Glomerular sieving coefficients were measured in anesthetized rats using neutral and anionic polysucrose during baseline conditions and glucagon-induced hyperfiltration. Data were analyzed using aligned-rank two-factor ANOVA, nonlinear mixed-effects regression of an electrostatic distributed two-pore model, pairwise correlation analysis, and principal component analysis. Results Hyperfiltration reduced the sieving of small and intermediate polysucrose molecules, whereas anionic polysucrose exhibited lower sieving coefficients than neutral polysucrose over a broad range of molecular sizes. An electrostatic distributed two-pore model accurately reproduced the observed effects of filtration rate and molecular charge and yielded an effective pore-wall charge density of 5.4 mC/m2 (95% confidence interval, 4.5 to 6.6). Pairwise correlation analysis revealed strong coupling between neighboring molecular sizes throughout the entire measured size range. Principal component analysis of the 2.5-8.0 nm size-selective region showed that the first principal component explained 96.3% of the variance and the first two principal components explained 99.9% of the variance. Separate analyses of the 2.5-5.0 nm and 5.0-8.0 nm transport regions showed that the first principal component explained 99.4% and 89.5% of the variance, respectively. Conclusions Glomerular sieving curves exhibited a highly constrained low-dimensional structure despite differences in molecular charge, filtration rate, and individual animals. The observed transport structure was consistent with distinct small-pore and large-pore transport domains and enabled highly effective principal component-based denoising of experimental sieving data.

Intrinsic lateral olivocochlear (iLOC) neurons provide vital brainstem efferent feedback to the cochlea in order to modulate hearing sensitivity through synapses onto type-I spiral ganglion neurons. During ageing or mutations affecting hair cell transduction in mice, efferent neurons rewire to form direct axo-somatic synapses onto inner hair cells (IHCs), recapitulating a synaptic configuration typically only restricted to the immature cochlea. Whether this rewiring reflects a compensatory mechanism or some form of attempted repair, or how iLOC biophysics change throughout ageing and this rewiring process, is not known. We utilised whole-cell patch-clamp electrophysiology to investigate iLOC activity and their underlying biophysics across the wild-type mouse lifespan. We show that iLOC neurons undergo a progressive increase in excitability with post-natal development and ageing, producing more spikes for a given stimulus. This intrinsic excitability shift was driven by the developmental decline in the A-type Kv4 mediated potassium current and increase in Kv2 mediated current. In ageing animals, and distinct from post-natal development, further increased firing rates were supported by an increased size of the fast-activating Kv3 current. Spontaneous bursting activity remained present in ageing iLOC neurons, and no reversion to an immature biophysics profile was evident. Interestingly, despite robustly eliciting efferent rewiring of IHCs, an accelerated ageing-like re-innervation genetic model did not recreate the biophysical changes in the iLOC neurons that reflected the ageing system. This work reveals distinct processes occurring within the iLOC feedback system, and shows that age-related enhancements of SGN resting activity are not triggered by deficits in IHC transduction.

This study tests the hypothesis that vasomotion, an [~] 0.1 Hz oscillation in arteriole diameter, is generated by intrinsic oscillations within the arterioles that perfuse the brain, and not by external drive from systemic blood pressure oscillations (Mayer waves). During cardio-pulmonary bypass that transiently eliminated systemic blood pressure oscillations in 14 patients, we observed that vasomotor oscillations persist with normal amplitudes and frequencies over the one- to three-hour time course of surgery. In contrast, [~] 0.1 Hz oscillations in peripheral blood pressure were predominantly absent. This implies that cerebral arterioles generate their own rhythmic vaso-dynamics, although we cannot discount that vasomotion can phase-lock with [~] 0.1 Hz systemic physiological rhythms in the awake, healthy state. We discuss the impact of this finding on the role of vasomotion in modulating the perfusion of blood and the transport of interstitial fluid in the brain.

Predictions of skeletal muscle energy consumption under a diverse range of muscle contractile conditions are critical for improving our understanding of locomotion. Existing mathematical models, while capturing the mechanical dependence of energy consuming processes, neglect the time-dependent behaviour and recovery costs associated with regenerating ATP. This time-dependence is important for predicting the energetic response of muscles during repetitive or cyclical tasks like locomotion, where muscle undergoes many contraction cycles. This study presents a novel model to predict energetic rates based on physiological processes: Ca2+ transport costs, cross-bridge cycling costs, and ATP regeneration. Previous mathematical models include the dependence on Ca2+ transport and cross-bridge cycling, but neglect the time-dependent response and the subsequent recovery of ATP following the contraction. Model parameters were obtained from existing data on isolated muscle preparations, and predicted energetic rates were validated on separate datasets across a range of contractile conditions including dynamic, sub-maximal, and twitch contractions. The time-dependent model was able to capture the influence of contraction frequency on peak energetic rates and the time-course of energetic recovery observed experimentally. The model captures key physiological processes while maintaining a minimal number of free parameters and low computational cost. This enables generalisability across muscles and species, and implementation into larger scale musculoskeletal models.

Synthetic opioids like fentanyl are a leading cause of overdose mortality. Although the hallmark of fentanyl overdose is ventilatory depression, fentanyl also induces tonic activation of skeletal musculature, including the diaphragm, which may advance progression of overdose towards death. While tonicity may further restrict diaphragmatic contractility, phase-specific dysregulation may also reflect a larger state of discoordination in respiratory control. Using urethane-anesthetized mice exposed to fentanyl, we test the hypothesis that fentanyl-induced diaphragm tonicity results from a loss of coordinated motor activity. Fentanyl produced two distinct phases: an initial phase of maximal ventilatory depression with preserved phasic activity, and a later phase characterized by unstable ventilation that partially rebounds, tonic diaphragmatic activation with loss of inspiratory phase dominance in EMG activity, and diminished bilateral diaphragmatic coordination. Carotid body denervation eliminated tonic activity and expiratory-phase EMG elevation, but it did not prevent hemi-diaphragm discoordination or ventilatory instability. Rhythmic brainstem slice recordings showed that bilateral preBotzinger complex burst-amplitude coupling was disrupted by u-opioid receptor (MOR) agonism. Furthermore, disordered diaphragm activity was reversed by administration of the MOR antagonist, Naloxone. Our findings reframe fentanyl overdose as a temporally evolving syndrome that involves distinct mechanisms to disrupt respiratory motor coordination.

Strikingly, highly trained athletes engaged in vertiginous activities (e.g., dance and slacklining) and patients with bilateral vestibular loss show a similar pattern of neural plasticity, likely resulting from reduced vestibular sensory processes. However, unlike patients, these athletes show no balance impairments, quite the opposite. This suggests that the attenuation of vestibular processing represents an adaptive recalibration to excessive vestibular stimulation rather than a sign of dysfunction. Concurrently, tactile processing increases as vestibular processing attenuates. Our findings indicate that effective adaptation extends beyond simple tactile compensation: it involves a strengthened tactile-brain pathway. Indeed, following unexpected base-of-support translations, the coupling between plantar shear forces (i.e., a proxy of plantar sole tactile afferents) and cortical responses over the somatosensory areas was markedly enhanced in Athletes. Cross-correlation analysis revealed stronger (r = 0.71) and faster (36 ms) tactile-brain coupling in Athletes (n = 25) compared with age- and gender-matched Controls (n = 18). This enhancement occurred within the first 180 ms following translation, that is, during the critical early phase of skin-surface interaction. Notably, artistic swimmers, who undergo intense vestibular stimulation in a weightless underwater environment without balance equilibrium constraints, also exhibit enhanced tactile-brain coupling. This suggests that strengthening the tactile-brain coupling is not merely a byproduct of balance expertise, but rather a broader adaptive response to sustained vestibular stimulation. Multimodal neurons integrating vestibular and somatosensory inputs, such as those in the somatosensory cortex and thalamus, may increase their responsiveness to foot tactile afferents when vestibular inputs become excessive. In such contexts, the somatosensory system may assume a dominant role in providing gravity-related information for balance control.

Sympathetic neuronal (SN) activity critically regulates the development and function of peripheral organs and tissues. Activity-dependent plasticity has been shown to modulate SN output, suggesting that compensatory forms of plasticity could contribute to maintaining stability of sympathetic circuits. Early SN hyperactivity drives the development of hypertension in humans and in the spontaneously hypertensive rat (SHR). In this study we used chemogenetic and pharmacological approaches, and took advantage of the enhanced activity of SHR SNs, to examine how long-term changes in activity impact synaptic properties in neonatal SN cultures. We showed that bidirectional changes in SN activity result in compensatory shifts in synaptic density that counteract long-term activity manipulations. These changes were mediated by satellite glial cells (SGCs), a non-neuronal cell in the sympathetic ganglia that has been shown to influence cholinergic synaptic sites during development. In the absence of SGCs there was no induction of homeostatic plasticity. Further, direct chemogenetic activation of SGCs was sufficient to drive compensatory plasticity, while glial inhibition blocked SN plasticity. We found that SGCs respond to cholinergic signaling by downregulating the expression of the synaptic regulators NGF and TNF, suggesting that neurons and glia interact to stabilize sympathetic output during long-term changes in circuit activity. Finally, we investigated whether these plasticity mechanisms are present in neonatal SHR SNs. We demonstrated that SHR SNs have an attenuated response to glia, both during synapse formation and activity-dependent plasticity. Taken together, this work outlines a novel homeostatic activity-dependent plasticity mechanism in the peripheral nervous system.