Reliability of resting-state functional connectivity in the human spinal cord: Assessing the impact of distinct noise sources.

The investigation of spontaneous fluctuations of the blood-oxygen-level-dependent (BOLD) signal has recently been extended from the brain to the spinal cord, where it has stimulated interest from a clinical perspective. A number of resting-state functional magnetic resonance imaging (fMRI) studies have demonstrated robust functional connectivity between the time series of BOLD fluctuations in bilateral dorsal horns and between those in bilateral ventral horns, in line with the functional neuroanatomy of the spinal cord. A necessary step prior to extension to clinical studies is assessing the reliability of such resting-state signals, which we aimed to do here in a group of 45 healthy young adults at the clinically prevalent field strength of 3T. When investigating connectivity in the entire cervical spinal cord, we observed fair to good reliability for dorsal-dorsal and ventral-ventral connectivity, whereas reliability was poor for within- and between-hemicord dorsal-ventral connectivity. Considering how prone spinal cord fMRI is to noise, we extensively investigated the impact of distinct noise sources and made two crucial observations: removal of physiological noise led to a reduction in functional connectivity strength and reliability - due to the removal of stable and participant-specific noise patterns - whereas removal of thermal noise considerably increased the detectability of functional connectivity without a clear influence on reliability. Finally, we also assessed connectivity within spinal cord segments and observed that while the pattern of connectivity was similar to that of whole cervical cord, reliability at the level of single segments was consistently poor. Taken together, our results demonstrate the presence of reliable resting-state functional connectivity in the human spinal cord even after thoroughly accounting for physiological and thermal noise, but at the same time urge caution if focal changes in connectivity (e.g. due to segmental lesions) are to be studied, especially in a longitudinal manner.

Cervical Epidural Electrical Stimulation Increases Respiratory Activity through Somatostatin-Expressing Neurons in the Dorsal Cervical Spinal Cord in Rats.

We tested the hypothesis that dorsal cervical epidural electrical stimulation (CEES) increases respiratory activity in male and female anesthetized rats. Respiratory frequency and minute ventilation were significantly increased when CEES was applied dorsally to the C2-C6 region of the cervical spinal cord. By injecting pseudorabies virus into the diaphragm and using c-Fos activity to identify neurons activated during CEES, we found neurons in the dorsal horn of the cervical spinal cord in which c-Fos and pseudorabies were co-localized, and these neurons expressed somatostatin (SST). Using dual viral infection to express the inhibitory Designer Receptors Exclusively Activated by Designer Drugs (DREADD), hM4D(Gi), selectively in SST-positive cells, we inhibited SST-expressing neurons by administering Clozapine N-oxide (CNO). During CNO-mediated inhibition of SST-expressing cervical spinal neurons, the respiratory excitation elicited by CEES was diminished. Thus, dorsal cervical epidural stimulation activated SST-expressing neurons in the cervical spinal cord, likely interneurons, that communicated with the respiratory pattern generating network to effect changes in ventilation.SIGNIFICANCE STATEMENT A network of pontomedullary neurons within the brainstem generates respiratory behaviors that are susceptible to modulation by a variety of inputs; spinal sensory and motor circuits modulate and adapt this output to meet the demands placed on the respiratory system. We explored dorsal cervical epidural electrical stimulation (CEES) excitation of spinal circuits to increase ventilation in rats. We identified dorsal somatostatin (SST)-expressing neurons in the cervical spinal cord that were activated (c-Fos-positive) by CEES. CEES no longer stimulated ventilation during inhibition of SST-expressing spinal neuronal activity, thereby demonstrating that spinal SST neurons participate in the activation of respiratory circuits affected by CEES. This work establishes a mechanistic foundation to repurpose a clinically accessible neuromodulatory therapy to activate respiratory circuits and stimulate ventilation.

Expression of Slit and Robo during remodeling of corticospinal tract in cervical spinal cord in middle cerebral artery occlusion rats.

BACKGROUND: Slits and Robos were associated with the generation of axons of corticospinal tract during the corticospinal tract (CST) remodeling after the cerebral ischemic stroke (CIS). However, little is known about the mechanism of CST remodeling. In this study, we detected the expression of Slits and Robos in middle cerebral artery occlusion (MCAO) rats to investigate the roles of Slits and Robos in the CIS. METHODS: MCAO model was established using modified Zea Longa method. Beam walking test (BWT) was conducted to evaluate the motor function. The images of the track of cortical spinal cord beam on day 7, 14 and 21 were observed by anterograde CST tracing. Biopinylated dextan amine (BDA) was used to mark CST anterogradely. Expression of GAP-43 mRNA and GAP-43 protein in cervical spinal cord was detected by Real-Time PCR and Western blot analysis, respectively. The expression of Slit1, Slit2 and Robo1 in cervical spinal cord was detected by immunofluorescence staining. RESULTS: The scores in the model group were significantly reduced compared to sham-operation group on day 7 (P < 0.001), 14 (P < 0.001) and 21 (P < 0.001), respectively. There was no significant difference in the score on day 7, 14 and 21 of the sham-operation groups (P > 0.05). In contrast, significant increase was noticed in the scores in model group, presenting a time-dependent manner. More CST staining fibers could be observed at the degenerative side in the model group compared with that of the sham-operation group on day 21. GAP-43 mRNA expression in the model group showed significant increase compared to that of sham-operation group on day 14 (P = 0.015) and 21 days (P = 0.002). The expression of GAP-43 protein in model group showed significant increase compared to that of sham-operation group on day 14 (P = 0.022) and day 21 (P = 0.008), respectively. The expression of Slit1 and Slit2 showed increase on day 14 and day 21, while the expression of Robo1 showed significant decrease in MCAO rats. CONCLUSION: Up-regulation of Slit1 and Slit2 and the downregulation of Robo1 may be related to the axons of CST midline crossing in spinal cord of MCAO rat during the spontaneous recovery of impaired motor function.

Vibration decreases the responsiveness of Ia afferents and spinal motoneurons in humans.

After vibration, Hoffmann reflex (H reflex) amplitude is depressed; however, the mechanisms underlying these phenomena remain unknown. This study investigated the influence of frequency and duration of vibration on the H reflex amplitude, heteronymous facilitation of the tendon jerk (T wave) mediated by group Ia afferents, and cervicomedullary motor evoked potential (CMEP) amplitude in 18 healthy human subjects. The H reflex of the flexor carpi radialis (FCR) was induced by median nerve stimulation at the elbow, and the conditioning FCR stimulation enhanced the T wave of the biceps brachii (BB). After vibration was applied to the FCR muscle belly, the amplitudes of the H reflex and heteronymous facilitation of the T wave were depressed; these influences persisted after the removal of vibration in all subjects. For the H reflex, there was no difference in the amount of depression among the frequencies of vibration used (57, 77, and 100 Hz). Higher frequencies of vibration were associated with longer recovery times of postvibration depression, and a longer duration of vibration was associated with a longer recovery time of the depression. Similar results were observed for heteronymous facilitation of the T wave, suggesting that the depression is caused by a decrease in postsynaptic potentials evoked by Ia afferents in spinal motoneurons; it was probably due to reduction in the number of Ia afferents recruited by the median nerve stimulation. Moreover, because the FCR CMEP amplitude was depressed after vibration, vibration should affect the responsiveness of spinal motoneurons. These mechanisms are considered to contribute to the H reflex depression after vibration.NEW & NOTEWORTHY Vibration decreased the responsiveness of Ia afferents from the muscle exposed to vibration, and the duration of depressive effect was modulated by the duration and frequency of the vibration: a longer duration and a higher frequency of vibration led to a longer recovery time of the depression. In addition to this presynaptic effect, it also depressed the responsiveness of spinal motoneurons, indicating postsynaptic inhibition through specific circuits triggered by Ia impulses.

Reductions in motoneuron excitability during sustained isometric contractions are dependent on stimulus and contraction intensity.

Cervicomedullary stimulation provides a means of assessing motoneuron excitability. Previous studies demonstrated that during low-intensity sustained contractions, small cervicomedullary evoked potentials (CMEPs) conditioned using transcranial magnetic stimulation (TMS-CMEPs) are reduced, whereas large TMS-CMEPs are less affected. As small TMS-CMEPs recruit motoneurons most active during low-intensity contractions whereas large TMS-CMEPs recruit a high proportion of motoneurons inactive during the task, these results suggest that reductions in motoneuron excitability could be dependent on repetitive activation. To further test this hypothesis, this study assessed changes in small and large TMS-CMEPs across low- and high-intensity contractions. Twelve participants performed a sustained isometric contraction of the elbow flexor for 4.5 min at the electromyography (EMG) level associated with 20% maximal voluntary contraction force (MVC; low intensity) and 70% MVC (high intensity). Small and large TMS-CMEPs with amplitudes of ∼15% and ∼50% Mmax at baseline, respectively, were delivered every minute throughout the tasks. Recovery measures were taken at 1-, 2.5- and 4-min postexercise. During the low-intensity trial, small TMS-CMEPs were reduced at 2-4 min (P ≤ 0.049) by up to -10% Mmax, whereas large TMS-CMEPs remained unchanged (P ≥ 0.16). During the high-intensity trial, small and large TMS-CMEPs were reduced at all time points (P < 0.01) by up to -14% and -33% Mmax, respectively, and remained below baseline during all recovery measures (P ≤ 0.02). TMS-CMEPs were unchanged relative to baseline during recovery following the low-intensity trial (P ≥ 0.24). These results provide novel insight into motoneuron excitability during and following sustained contractions at different intensities and suggest that contraction-induced reductions in motoneuron excitability depend on repetitive activation.NEW & NOTEWORTHY This study measured motoneuron excitability using cervicomedullary evoked potentials conditioned using transcranial magnetic stimulation (TMS-CMEPs) of both small and large amplitudes during sustained low- and high-intensity contractions of the elbow flexors. During the low-intensity task, only the small TMS-CMEP was reduced. During the high-intensity task, both small and large TMS-CMEPs were substantially reduced. These results indicate that repetitively active motoneurons are specifically reduced in excitability compared with less active motoneurons in the same pool.

Effect of acute intermittent hypoxia on cortico-diaphragmatic conduction in healthy humans.

Acute intermittent hypoxia (AIH) is a strategy to improve motor output in humans with neuromotor impairment. A single AIH session increases the amplitude of motor evoked potentials (MEP) in a finger muscle (first dorsal interosseous), demonstrating enhanced corticospinal neurotransmission. Since AIH elicits phrenic/diaphragm long-term facilitation (LTF) in rodent models, we tested the hypothesis that AIH augments diaphragm MEPs in humans. Eleven healthy adults (7 males, age = 29 ± 6 years) were tested. Transcranial and cervical magnetic stimulation were used to induce diaphragm MEPs and compound muscle action potentials (CMAP) recorded by surface EMG, respectively. Stimulus-response curves were generated prior to and 30-60 min after AIH. Diaphragm LTF was assessed by measurement of integrated EMG burst amplitude and frequency during eupnoeic breathing before and after AIH. Following baseline measurements, AIH was delivered from an oxygen generator connected to a facemask under poikilocapnic conditions (15 one minute episodes of 9% inspired oxygen with one minute room air intervals). There were no detectable changes in MEP (-1.5 ± 12.1%, p = 0.96) or CMAP (+0.1 ± 7.8%, p = 0.97) amplitudes across the stimulus-response curve. At stimulation intensities approximating 50% of the difference between minimum and maximum baseline amplitudes, MEP and CMAP amplitudes were also unchanged (p > 0.05). Further, no AIH effect was observed on diaphragm EMG activity during eupnoea post-AIH (p > 0.05). We conclude that unlike hand muscles, poikilocapnic AIH does not enhance diaphragm MEPs or produce diaphragm LTF in healthy humans.

Noninvasive measurement of sensory action currents in the cervical cord by magnetospinography.

OBJECTIVE: To obtain magnetic recordings of electrical activities in the cervical cord and visualize sensory action currents of the dorsal column, intervertebral foramen, and dorsal horn. METHODS: Neuromagnetic fields were measured at the neck surface upon median nerve stimulation at the wrist using a magnetospinography system with high-sensitivity superconducting quantum interference device sensors. Somatosensory evoked potentials (SEPs) were also recorded. Evoked electrical currents were reconstructed by recursive null-steering beamformer and superimposed on cervical X-ray images. RESULTS: Estimated electrical currents perpendicular to the cervical cord ascended sequentially. Their peak latency at C5 and N11 peak latency of SEP were well-correlated in all 16 participants (r = 0.94, p < 0.0001). Trailing axonal currents in the intervertebral foramens were estimated in 10 participants. Estimated dorsal-ventral electrical currents were obtained within the spinal canal at C5. Current density peak latency significantly correlated with cervical N13-P13 peak latency of SEPs in 13 participants (r = 0.97, p < 0.0001). CONCLUSIONS: Magnetospinography shows excellent spatial and temporal resolution after median nerve stimulation and can identify the spinal root entry level, calculate the dorsal column conduction velocity, and analyze segmental dorsal horn activity. SIGNIFICANCE: This approach is useful for functional electrophysiological diagnosis of somatosensory pathways.

A TrkB Antibody Agonist Promotes Plasticity after Cervical Spinal Cord Injury in Adult Rats.

After spinal cord injury (SCI) in mammals, there is only limited repair and, consequently, only moderate recovery. One mechanism frequently discussed to be involved in this recovery is plasticity (i.e., adaptations in spared neuronal circuitries). In the current study, we tested the effect of an intrathecal application of the TrkB agonist antibody, 29D7, on plasticity after cervical SCI in adult rats. Treatment with 29D7 for 4 weeks resulted in an ∼50% increase in collateral sprouting of severed corticospinal tract fibers above the lesion compared to the control group and enhanced branching in the gray matter rostral to the injury. Growth-associated protein 43 immunoreactivity in the spinal cord rostral to the level of the injury as well as contralateral to the lesion was also increased. These indications of enhanced plasticity by 29D7 were paralleled by improved performances of the mildly affected paw, as assessed by Montoya and tray reaching tasks. The reaching behaviors of the paw ipsilateral to the side of severe injury to the cortico- and rubrospinal tract were not altered by the treatment. The present study suggests that 29D7 may be a potential candidate to promote plasticity and functional recovery, especially after moderate SCI. Future studies confirming these results, along with a potential combinatory therapy including rehabilitative training, will be needed to evaluate the clinical value of such a treatment.

Forelimb force direction and magnitude independently controlled by spinal modules in the macaque.

Modular organization of the spinal motor system is thought to reduce the cognitive complexity of simultaneously controlling the large number of muscles and joints in the human body. Although modular organization has been confirmed in the hindlimb control system of several animal species, it has yet to be established in the forelimb motor system or in primates. Expanding upon experiments originally performed in the frog lumbar spinal cord, we examined whether costimulation of two sites in the macaque monkey cervical spinal cord results in motor activity that is a simple linear sum of the responses evoked by stimulating each site individually. Similar to previous observations in the frog and rodent hindlimb, our analysis revealed that in most cases (77% of all pairs) the directions of the force fields elicited by costimulation were highly similar to those predicted by the simple linear sum of those elicited by stimulating each site individually. A comparable simple summation of electromyography (EMG) output, especially in the proximal muscles, suggested that this linear summation of force field direction was produced by a spinal neural mechanism whereby the forelimb motor output recruited by costimulation was also summed linearly. We further found that the force field magnitudes exhibited supralinear (amplified) summation, which was also observed in the EMG output of distal forelimb muscles, implying a novel feature of primate forelimb control. Overall, our observations support the idea that complex movements in the primate forelimb control system are made possible by flexibly combined spinal motor modules.

Visualization of electrical activity in the cervical spinal cord and nerve roots after ulnar nerve stimulation using magnetospinography.

OBJECTIVE: To establish a method for magnetospinography (MSG) measurement after ulnar nerve stimulation and to clarify its characteristics. METHODS: Using a 132-channel magnetoneurography system with a superconducting quantum interference device, cervical MSG measurements were obtained for 10 healthy volunteers after stimulation of the ulnar nerve at the elbow and the wrist, and neural current distribution was calculated and superimposed on the cervical X-ray images. RESULTS: Neuromagnetic signals were obtained in all participants after applying the stimulus artifact removal algorithm. The measured magnetic field intensity after elbow stimulation was about twice that after wrist stimulation. Calculated neural currents flowed into the intervertebral foramina at C6/7 to T1/2 and propagated cranially along the spinal canal. The conduction velocity from the peak latency of inward currents at C5-C7 was 73.4 ± 19.6 m/s. CONCLUSIONS: We successfully obtained MSG measurements after ulnar nerve stimulation. The neural currents flowed into the spinal canal from more caudal segments after ulnar nerve stimulation compared with median nerve stimulation, and these MSG measurements were effective in examining the spinal tracts at C5/6/7. SIGNIFICANCE: This is the first report on the use of MSG to visualize electrical activity in the cervical spinal cord and nerve root after ulnar nerve stimulation.

A Randomized Sham-Controlled Cross-Over Study on the Short-Term Effect of Non-Invasive Cervical Vagus Nerve Stimulation on Spinal and Supraspinal Nociception in Healthy Subjects.

OBJECTIVE: The aim of the present study was to test the effects of vagus nerve stimulation (VNS) on the descending pain inhibition, quantified by the nociceptive flexor (RIII) reflex and the conditioned pain modulation (CPM) paradigm, and on supraspinal nociceptive responses, assessed by pain intensity and unpleasantness ratings and late somatosensory evoked potentials (SEPs), in healthy subjects. BACKGROUND: Non-invasive vagus nerve stimulation (nVNS) showed promising effects on headache and pain treatment. Underlying mechanisms are only incompletely understood but may include the activation of the descending pain inhibitory system and/or the modification of emotional responses to pain. METHODS: Twenty-seven adult, healthy, and pain-free subjects participated in this double-blind cross-over study conducted at a university research center. They received 4 minutes of cervical nVNS or sham stimulation in randomized order. RIII reflexes, pain ratings, and SEPs were assessed before, during, and 5, 15, 30, and 60 minutes after nVNS/sham stimulation, followed by CPM testing. The primary outcome was the nVNS effect on the RIII reflex size. Three subjects were excluded after the preparatory session (before randomization), 1 subject was excluded after outlier analysis, leaving 23 for analysis. RESULTS: RIII reflex areas were 917.1 ± 563.8 µV × ms (mean ± SD) before, 952.4 ± 467.4 µV × ms during and 929.2 ± 484.0 µV × ms immediately after nVNS and 858.4 ± 489.2 µV × ms before, 913.9 ± 539.7 µV × ms during and 862.4 ± 476.0 µV × ms after sham stimulation, revealing no differences between the immediate effects of nVNS and sham stimulation (F [3,66]  = 0.67, P = .574). There also were no effects of nVNS over sham on RIII reflex areas up to 60 minutes after nVNS (F [1.7,37.4]  = 1.29, P = .283). Similarly, there was no statistically significant effect of nVNS on pain intensity ratings and thresholds, RIII reflex thresholds, late SEP amplitudes, and the CPM effect, compared to sham. Pain unpleasantness ratings statistically significantly decreased from 4.4 ± 2.4 (NRS 0-10) to 4.1 ± 2.5 during nVNS compared to sham stimulation (F [1,22]  = 8.74, P = .007), but there were no longer lasting effects (5-60 minutes after stimulation). CONCLUSIONS: The present study does not support an acute effect of nVNS on descending pain inhibition, pain intensity perception or supraspinal nociception in healthy adults. However, there was a small effect on pain unpleasantness during nVNS, suggesting that nVNS may preferentially act on affective, not somatosensory pain components.

Assessing the spatial distribution of cervical spinal cord activity during tactile stimulation of the upper extremity in humans with functional magnetic resonance imaging.

Dermatomal maps are a mainstay of clinical practice and provide information on the spatial distribution of the cutaneous innervation of spinal nerves. Dermatomal deficits can help isolate the level of spinal nerve root involvement in spinal conditions and guide clinicians in diagnosis and treatment. Dermatomal maps, however, have limitations, and the spatial distribution of spinal cord sensory activity in humans remains to be quantitatively assessed. Here we used spinal cord functional MRI to map and quantitatively compare the spatial distribution of sensory spinal cord activity during tactile stimulation of the left and right lateral shoulders (i.e. C5 dermatome) and dorsal third digits of the hands (i.e., C7 dermatome) in healthy humans (n â€‹= â€‹24, age â€‹= â€‹36.8 â€‹± â€‹11.8 years). Based on the central sites for processing of innocuous tactile sensory information, we hypothesized that the activity would be localized more to the ipsilateral dorsal spinal cord with the lateral shoulder stimulation activity being localized more superiorly than the dorsal third digit. The findings demonstrate lateralization of the activity with the left- and right-sided stimuli having more activation in the ipsilateral hemicord. Contradictory to our hypotheses, the activity for both stimulation sites was spread across the dorsal and ventral hemicords and did not demonstrate a clear superior-inferior localization. Instead, the activity for both stimuli had a broader than expected distribution, extending across the C5, C6, and C7 spinal cord segments. We highlight the complexity of the human spinal cord neuroanatomy and several sources of variability that may explain the observed patterns of activity. While the findings were not completely consistent with our a priori hypotheses, this study provides a foundation for continued work and is an important step towards developing normative quantitative spinal cord measures of sensory function, which may become useful objective MRI-based biomarkers of neurological injury and improve the management of spinal disorders.

Stimulus outputs induced by subdural electrodes on the cervical spinal cord in monkeys.

OBJECTIVE: Spinal stimulation is a promising method for restoring the function of paralyzed limbs following neurological damage to descending pathways. The present study examined the forelimb movements and muscle responses evoked by subdural spinal stimulation of the cervical cord in sedated monkeys or during an arm-reaching task. APPROACH: We chronically implanted a platinum subdural electrode array with eight channels over the dorsal-lateral aspect of the cervical enlargement. The electrodes had a diameter of 1 mm and an inter-electrode center-to-center distance of 3 mm. Subdural spinal micro-stimulation was delivered at sites while the monkeys were sedated or performed arm-reaching movements. MAIN RESULTS: The evoked movements clearly showed the somatotopic map of the output sites; the electrodes located on the rostral cervical cord tended to induce movements of the proximal arm, whereas the caudal electrodes tended to induce movements of the distal joints, such as the wrist and digits. To document the muscle responses evoked by subdural spinal stimulation, stimulus-triggered averages of rectified electromyograms were compiled when the monkeys performed an arm-reaching task or were sedated. Under sedation, evoked facilitative muscle responses were observed in vicinity muscles. In contrast, during the task, stimulation evoked facilitative or suppressive responses in multiple muscles, including those located on proximal and distal joints, while somatotopy became blurred under sedation. Furthermore, stimulation during tasks activated synergistic muscle groups. For example, stimuli strongly facilitated finger extensor muscles, but suppressed the antagonist muscles. SIGNIFICANCE: These dynamic changes in muscle representation by subdural cervical spinal stimulation between sedated and awake states help our understanding of the nature of spinal circuits and will facilitate the development of neuroprosthetic technology to regain motor function after neural damage to the descending pathways.

Effect of blood flow occlusion on corticospinal excitability during sustained low-intensity isometric elbow flexion.

Blood flow occlusion (BFO) has been used to study the influence of group III/IV muscle afferents after fatiguing exercise, but it is unknown how BFO-induced activity of these afferents affects motor cortical and motoneuronal excitability during low-intensity exercise. Therefore, the purpose of this study was to assess the acute effect of BFO on peripheral [maximal M wave (Mmax)], spinal [cervicomedullary motor evoked potential (CMEP) normalized to Mmax], and motor cortical [motor evoked potential (MEP) normalized to CMEP] excitability. Nine healthy men completed a sustained isometric contraction of the elbow flexors at 20% of maximal force under three conditions: 1) contractile failure with BFO, 2) a time-matched trial without restriction [free flow (FFiso)], and 3) contractile failure with free flow (FFfail). Time to failure for BFO (and FFiso) were ~80% shorter than that for FFfail (P < 0.05). For FFfail and FFiso, Mmax area decreased ~17% and ~7%, respectively (P < 0.05), with no change during BFO. CMEP/Mmax area increased ~226% and ~80% during BFO and FFfail, respectively (P < 0.05), with no change during FFiso (P > 0.05). The increase in normalized CMEP area was greater for BFO and FFfail compared with FFiso and for BFO compared with FFfail. MEP/CMEP area was not different among the protocols (P > 0.05) and increased ~64% with time (P < 0.05). It is likely that group III/IV muscle afferent feedback to the spinal cord modulates the large increase in motoneuronal excitability for the BFO compared with FFfail and FFiso protocols.NEW & NOTEWORTHY We have observed how blood flow occlusion modulates motor cortical, spinal, and peripheral excitability during and immediately after a sustained low-intensity isometric elbow flexion contraction to failure. We conclude that blood flow occlusion causes a greater and more rapid increase in motoneuronal excitability.

Mid-cervical interneuron networks following high cervical spinal cord injury.

Spinal interneuron (IN) networks can facilitate respiratory motor recovery after spinal cord injury (SCI). We hypothesized that excitatory synaptic connectivity between INs located immediately caudal to unilateral cervical SCI would be most prevalent in a contra- to ipsilateral direction. Adult rats were studied following chronic C2 spinal cord hemisection (C2Hx) injury. Rats were anesthetized and ventilated and a multi-electrode array was used to simultaneously record INs on both sides of the C4-5 spinal cord. The temporal firing relationship between IN pairs was evaluated using cross-correlation with directionality of synaptic connections inferred based on electrode location. During baseline recordings, the majority of detectable excitatory IN connections occurred in a contra- to- ipsilateral direction. However, acute respiratory stimulation with hypoxia abolished this directionality, while simultaneously increasing the detectable inhibitory connections within the ipsilateral cord. We conclude that propriospinal networks caudal to SCI can display a contralateral-to-ipsilateral directionality of synaptic connections and that these connections are modulated by acute exposure to hypoxia.

The effects of forearm position and contraction intensity on cortical and spinal excitability during a submaximal force steadiness task of the elbow flexors.

Elbow flexor force steadiness is less with the forearm pronated (PRO) compared with neutral (NEU) or supinated (SUP) and may relate to neural excitability. Although not tested in a force steadiness paradigm, lower spinal and cortical excitability was observed separately for biceps brachii in PRO, possibly dependent on contractile status at the time of assessment. This study aimed to investigate position-dependent changes in force steadiness as well as spinal and cortical excitability at a variety of contraction intensities. Thirteen males (26 ± 7 yr; means ± SD) performed three blocks (PRO, NEU, and SUP) of 24 brief (~6 s) isometric elbow flexor contractions (5, 10, 25 or 50% of maximal force). During each contraction, transcranial magnetic stimulation or transmastoid stimulation was delivered to elicit a motor-evoked potential (MEP) or cervicomedullary motor-evoked potential (CMEP), respectively. Force steadiness was lower in PRO compared with NEU and SUP (P ≤ 0.001), with no difference between NEU and SUP. Similarly, spinal excitability (CMEP/maximal M wave) was lower in PRO than NEU (25 and 50% maximal force; P ≤ 0.010) and SUP (all force levels; P ≤ 0.004), with no difference between NEU and SUP. Cortical excitability (MEP/CMEP) did not change with forearm position (P = 0.055); however, a priori post hoc testing for position showed excitability was 39.8 ± 38.3% lower for PRO than NEU at 25% maximal force (P = 0.006). The data suggest that contraction intensity influences the effect of forearm position on neural excitability and that reduced spinal and, to a lesser extent, cortical excitability could contribute to lower force steadiness in PRO compared with NEU and SUP.NEW & NOTEWORTHY To address conflicting reports about the effect of forearm position on spinal and cortical excitability of the elbow flexors, we examine the influence of contraction intensity. For the first time, excitability data are considered in a force steadiness context. Motoneuronal excitability is lowest in pronation and this disparity increases with contraction intensity. Cortical excitability exhibits a similar pattern from 5 to 25% of maximal force. Lower corticospinal excitability likely contributes to relatively poor force steadiness in pronation.

Somatosensory corticospinal tract axons sprout within the cervical cord following a dorsal root/dorsal column spinal injury in the rat.

The corticospinal tract (CST) is the major descending pathway controlling voluntary hand function in primates, and though less dominant, it mediates voluntary paw movements in rats. As with primates, the CST in rats originates from multiple (albeit fewer) cortical sites, and functionally different motor and somatosensory subcomponents terminate in different regions of the spinal gray matter. We recently reported in monkeys that following a combined cervical dorsal root/dorsal column lesion (DRL/DCL), both motor and S1 CSTs sprout well beyond their normal terminal range. The S1 CST sprouting response is particularly dramatic, indicating an important, if poorly understood, somatosensory role in the recovery process. As rats are used extensively to model spinal cord injury, we asked if the S1 CST response is conserved in rodents. Rats were divided into sham controls, and two groups surviving post-lesion for ~6 and 10 weeks. A DRL/DCL was made to partially deafferent one paw. Behavioral testing showed a post-lesion deficit and recovery over several weeks. Three weeks prior to ending the experiment, S1 cortex was mapped electrophysiologically, for tracer injection placement to determine S1 CST termination patterns within the cord. Synaptogenesis was also assessed for labeled S1 CST terminals within the dorsal horn. Our findings show that the affected S1 CST sprouts well beyond its normal range in response to a DRL/DCL, much as it does in macaque monkeys. This, along with evidence for increased synaptogenesis post-lesion, indicates that CST terminal sprouting following a central sensory lesion, is a robust and conserved response.

Muscle activation pattern elicited through transcutaneous stimulation near the cervical spinal cord.

OBJECTIVE: Neuromuscular electrical stimulation can help activate muscles of individuals with neurological disorders. However, conventional electrical stimulation targets distal branches of motor axons, and activates muscles non-physiologically. For example, stimulation at the muscle belly activates muscles in a highly synchronized manner. Accordingly, we investigated whether the muscle activation pattern was more asynchronized through transcutaneous stimulation near the cervical spinal cord (tsCSC). APPROACH: A stimulation array was placed on the posterior side near the cervical spinal cord, to target the arm and hand muscles. Stimulation trains of 10 Hz and 30 Hz were delivered. Electromyogram signals were recorded to quantify the muscle activation patterns. Arm and finger joint kinematics were also recorded using a motion capture system. MAIN RESULTS: After an initial synchronized activation prior to 35 ms after stimulation onset, we observed substantial asynchronized muscle activities with a long latency (>35 ms). The asynchronized activation is also more evident in distal muscles compared with the proximal muscles. In addition, the decreased synchronization level of muscle activities correlated with a reduced fluctuation of joint movement. The highly asynchronized muscle activities indicated an activation of the sensory axons and/or dorsal roots as well as a possible involvement of some spinal-supraspinal circuitry. SIGNIFICANCE: Our tsCSC approach can improve the muscle activation pattern during electrical stimulation with a possible involvement of the spinal and supraspinal pathways, which can facilitate applications on rehabilitation/assistance of individuals with impaired motor function.

Respiratory resetting elicited by single pulse spinal stimulation.

Intraspinal microstimulation (ISMS) can effectively activate spinal motor circuits, but the impact on the endogenous respiratory pattern has not been systematically evaluated. Here we delivered ISMS in spontaneously breathing adult rats while simultaneously recording diaphragm and external intercostal electromyography activity. ISMS pulses were delivered from C2-T1 along two rostrocaudal tracts located 0.5 or 1 mm lateral to midline. A tungsten electrode was incrementally advanced from the dorsal spinal surface and 300µs biphasic pulses (10-90 µA) were delivered at depth increments of 600 µm. Dorsal ISMS often produced fractionated inspiratory bursting or caused early termination of the inspiratory effort. Conversely, ventral stimulation had no discernable impact on respiratory resetting. We conclude that ISMS targeting the ventral spinal cord is unlikely to directly alter the respiratory rhythm. Dorsal ISMS, however, may terminate the inspiratory burst through activation of spinobulbar pathways. We suggest that respiratory patterns should be included as an outcome variable in preclinical studies of ISMS.

Transcutaneous spinal cord stimulation of the cervical cord modulates lumbar networks.

It has been established that coordinated arm and leg (A&L) cycling facilitates corticospinal drive and modulation of cervico-lumbar connectivity and ultimately improves overground walking in people with incomplete spinal cord injury or stroke. This study examined the effect of noninvasive transcutaneous spinal cord stimulation (tSCS) on the modulation of cervico-lumbar connectivity. Thirteen neurologically intact adults participated in the study. The excitability of the Hoffmann (H) reflex elicited in the soleus muscle was examined under multiple conditions involving either the arms held in a static position or rhythmic arm cycling while tSCS was applied to either the cervical or lumbar cord. As expected, soleus H-reflex amplitude was significantly suppressed by 19.2% during arm cycling (without tSCS) relative to arms static (without tSCS). Interestingly, tSCS of the cervical cord with arms static significantly suppressed the soleus H-reflex (-22.9%), whereas tSCS over the lumbar cord did not suppress the soleus H-reflex (-3.8%). The combination of arm cycling with cervical or lumbar tSCS did not yield additional suppression of the soleus H-reflex beyond that obtained with arm cycling alone or cervical tSCS alone. The results demonstrate that activation of the cervical spinal cord through both rhythmic arm cycling and tonic tSCS significantly modulates the activity of lumbar networks. This highlights the potential for engaging cervical spinal cord networks through tSCS during rehabilitation interventions to enhance cervico-lumbar connectivity. This connectivity is influential in facilitating improvements in walking function after neurological impairment.NEW & NOTEWORTHY This is the first study to investigate the modulatory effects of transcutaneous spinal cord stimulation (tSCS) on cervico-lumbar connectivity. We report that both rhythmic activation of the cervical spinal cord through arm cycling and tonic activation of the cervical cord through tSCS significantly modulate the activity of lumbar networks. This suggests that engaging cervical spinal cord networks through tSCS during locomotor retraining interventions may not only enhance cervico-lumbar connectivity but also further improve walking capacity.