Post-activation depression of the Hoffman reflex is not altered by galvanic vestibular stimulation in healthy subjects.

The comprehension of the neural elements interacting in the spinal cord affected by vestibular input will contribute to the understanding of movement execution in normal and pathological conditions. In this context, Hoffman's reflex (H-reflex) has been used to evaluate transient excitability changes on the spinal cord descending pathways. The post-activation depression (P-AD) of the H-reflex consists of evoking consecutive responses (>1 Hz) provoking an amplitude depression, which has been shown to diminish in pathological conditions (i.e., spasticity, diabetic neuropathy). Galvanic Vestibular Stimulation (GVS) is a non-invasive method that activates the vestibular afferents and has been used to study the excitability of the H-reflex applied as a conditioning pulse. To our knowledge, there are no reports evaluating the P-AD during and after GVS. Our primary aim was to determine if GVS alters the P-AD evoked by stimulating the tibial nerve at 0.1, 1, 5, and 10 Hz, recording in the gastrocnemius and soleus muscles. Direct current stimulation of 2.0 ± 0.6 mA with the cathode ipsilateral (Ipsi) or contralateral (Contra) to the H-reflex electrode montage was applied bilaterally over the mastoid process in 19 healthy subjects. The P-AD's immediate post-GVS response (P Ipsi, P Contra) was also analyzed. Secondarily, we analyzed the excitability of the H-reflex during GVS. Responses evoked at 0.1 Hz with GVS, post-GVS, and a Control (no GVS) condition were used for comparisons. Our results show that P-AD persisted in all subjects despite increased excitability induced by GVS: statistical significance was found when comparing P-AD at 1, 5, and 10 Hz with the corresponding condition (Control, Ipsi, P Ipsi, Contra, P Contra) at 0.1 Hz (p < 0.001). Additionally, the increase in excitability produced by GVS was quantified for the first H-reflex of each P-AD stimulation frequency. The percentage change for all GVS conditions surpassed the Control by at least 20%, being statistically significant for Contra compared to Control (p < 0.01). In summary, although GVS increases the excitability of the vestibulospinal pathway at a premotor level, the neural inhibitory mechanism present in P-AD remains unaltered in healthy subjects.

Evidence of impaired H-reflex and H-reflex rate-dependent depression in diabetes, prediabetes and obesity: a mini-review.

Diabetes Mellitus is a public health problem associated with complications such as neuropathy; however, it has been proposed that these may begin to develop during prediabetes and may also be present in persons with obesity. Diabetic peripheral neuropathy is the presence of signs and/or symptoms of peripheral nerve dysfunction in people living with diabetes, which increases the risk of developing complications and has a deleterious impact on quality of life. As part of the therapeutic protocol for diabetes, screening tests to identify peripheral neuropathy are suggested, however, there are no recommendations for people with prediabetes and obesity without symptoms such as pain, numbness, or paresthesias. Moreover, clinical screening tests that are usually used to recognize this alteration, such as tendon reflex, temperature sensation, and pressure and vibration perception, might be subjective as they depend on the evaluator's experience thus the incorrect application of these tests may not recognize the damage to small or large-nerve fibers. Recent evidence suggests that an objective study such as the impairment of the rate-dependent depression of the H-reflex could be used as a biomarker of spinal disinhibition and hence may provide more information on sensorimotor integration.

Impaired rate-dependent depression of the H-reflex in type-2 diabetes, prediabetes, overweight and obesity: A cross-sectional study.

Type-2 diabetes is a chronic metabolic disorder characterized by hyperglycemia, resulting from deficits in insulin secretion or insulin resistance. According to the International Diabetes Federation, approximately 463 million people suffered from this condition in 2019, with a rapidly increasing impact in low-and middle-income countries. Obesity is a well-known risk factor for diabetes, and current data project a continuous increase in diabetes prevalence worldwide in obese individuals. Among the common complications, diabetic peripheral neuropathy (DPN) causes sensory symptoms, including pain that contributes to foot ulceration, and if not controlled, limb amputation may occur. The diagnosis of DPN is a clinical problem. Rate-dependent depression (RDD) of the Hoffmann reflex in the lower limbs has been proposed as a test to determine the presence of neuropathic pain in subjects with type-1 and type-2 diabetes. Recently, impaired RDD has been described in obese and diabetic rodent models. In this study, we characterized the RDD by evaluating the H-reflex at 0.2, 1, 2, 5, and 10 Hz in 39 patients with type-2 Diabetes mellitus (T2DM) and 42 controls without diabetes, subsequently classified as overweight/obese and prediabetic. A significant decrease in the RDD of the H-reflex was found in T2DM subjects at 1, 2, 5, and 10 Hz (P < .001) stimulation frequencies compared to controls, but not at 0.2 Hz (P = .48). A major finding of this study is that impaired RDD was also found in 11/25 overweight and obese subjects in at least 2 stimulation frequencies, being 10 of those classified in prediabetic levels according to their HbA1C values. The RDD of the H-reflex could be used as a quantitative and sensitive tool to study T2DM subpopulations with peripheral neuropathy. RDD could be used as a screening tool in combination with clinical tests to diagnose DPN and evaluate the progression of this condition.

Newly regenerated axons via scaffolds promote sub-lesional reorganization and motor recovery with epidural electrical stimulation.

Here, we report the effect of newly regenerated axons via scaffolds on reorganization of spinal circuitry and restoration of motor functions with epidural electrical stimulation (EES). Motor recovery was evaluated for 7 weeks after spinal transection and following implantation with scaffolds seeded with neurotrophin producing Schwann cell and with rapamycin microspheres. Combined treatment with scaffolds and EES-enabled stepping led to functional improvement compared to groups with scaffold or EES, although, the number of axons across scaffolds was not different between groups. Re-transection through the scaffold at week 6 reduced EES-enabled stepping, still demonstrating better performance compared to the other groups. Greater synaptic reorganization in the presence of regenerated axons was found in group with combined therapy. These findings suggest that newly regenerated axons through cell-containing scaffolds with EES-enabled motor training reorganize the sub-lesional circuitry improving motor recovery, demonstrating that neuroregenerative and neuromodulatory therapies cumulatively enhancing motor function after complete SCI.

Supraspinal and Afferent Signaling Facilitate Spinal Sensorimotor Network Excitability After Discomplete Spinal Cord Injury: A Case Report.

OBJECTIVE: In this study, we evaluated the role of residual supraspinal and afferent signaling and their convergence on the sublesional spinal network in subject diagnosed with complete paralysis (AIS-A). METHODS: A combination of electrophysiologic techniques with positional changes and subject-driven reinforcement maneuvers was implemented in this study. Electrical stimulation was applied transcutaneously at the T9-L2 vertebra levels and the spinal cord motor evoked potentials (SEMP) were recorded from leg muscles. To test the influence of positional changes, the subject was placed in (i) supine, (ii) upright with partial body weight bearing and (iii) vertically suspended without body weight bearing positions. RESULTS: Increase in amplitude of SEMP was observed during transition from supine to upright position, supporting the role of sensory input in lumbosacral network excitability. Additionally, amplitudes of SEMP were facilitated during reinforcement maneuvers, indicating a supralesional influence on sub-lesional network. After initial assessment, subject underwent rehabilitation therapy with following electrophysiological testing that reviled facilitation of SEMP. CONCLUSION: These results demonstrate that combination of electrophysiological techniques with positional and reinforcement maneuvers can add to the diagnostics of discomplete SCI. These findings also support an idea that integration of supraspinal and afferent information on sub-lesional circuitry plays a critical role in facilitation of spinal sensorimotor network in discomplete SCI.

Changes in spinal cord hemodynamics reflect modulation of spinal network with different parameters of epidural stimulation.

In this study functional ultrasound (fUS) imaging has been implemented to explore the local hemodynamics response induced by electrical epidural stimulation and to study real-time in vivo functional changes of the spinal cord, taking advantage of the superior spatiotemporal resolution provided by fUS. By quantifying the hemodynamics and electromyographic response features, we tested the hypothesis that the temporal hemodynamics response of the spinal cord to electrical epidural stimulation could reflect modulation of the spinal circuitry and accordingly respond to the changes in parameters of electrical stimulation. The results of this study for the first time demonstrate that the hemodynamics response to electrical stimulation could reflect a neural-vascular coupling of the spinal cord. Response in the dorsal areas to epidural stimulation was significantly higher and faster compared to the response in ventral spinal cord. Positive relation between the hemodynamics and the EMG responses was observed at the lower frequencies of epidural stimulation (20 and 40 Hz), which according to our previous findings can facilitate spinal circuitry after spinal cord injury, compared to higher frequencies (200 and 500 Hz). These findings suggest that different mechanisms could be involved in spinal cord hemodynamics changes during different parameters of electrical stimulation and for the first time provide the evidence that neural-vascular coupling of the spinal cord circuitry could be related to specific organization of spinal cord vasculature and hemodynamics.

Multifactorial motor behavior assessment for real-time evaluation of emerging therapeutics to treat neurologic impairments.

Integrating multiple assessment parameters of motor behavior is critical for understanding neural activity dynamics during motor control in both intact and dysfunctional nervous systems. Here, we described a novel approach (termed Multifactorial Behavioral Assessment (MfBA)) to integrate, in real-time, electrophysiological and biomechanical properties of rodent spinal sensorimotor network activity with behavioral aspects of motor task performance. Specifically, the MfBA simultaneously records limb kinematics, multi-directional forces and electrophysiological metrics, such as high-fidelity chronic intramuscular electromyography synchronized in time to spinal stimulation in order to characterize spinal cord functional motor evoked potentials (fMEPs). Additionally, we designed the MfBA to incorporate a body weight support system to allow bipedal and quadrupedal stepping on a treadmill and in an open field environment to assess function in rodent models of neurologic disorders that impact motor activity. This novel approach was validated using, a neurologically intact cohort, a cohort with unilateral Parkinsonian motor deficits due to midbrain lesioning, and a cohort with complete hind limb paralysis due to T8 spinal cord transection. In the SCI cohort, lumbosacral epidural electrical stimulation (EES) was applied, with and without administration of the serotonergic agonist Quipazine, to enable hind limb motor functions following paralysis. The results presented herein demonstrate the MfBA is capable of integrating multiple metrics of motor activity in order to characterize relationships between EES inputs that modulate mono- and polysynaptic outputs from spinal circuitry which in turn, can be used to elucidate underlying electrophysiologic mechanisms of motor behavior. These results also demonstrate that proposed MfBA is an effective tool to integrate biomechanical and electrophysiology metrics, synchronized to therapeutic inputs such as EES or pharmacology, during body weight supported treadmill or open field motor activities, to target a high range of variations in motor behavior as a result of neurological deficit at the different levels of CNS.

Functional Ultrasound Imaging of Spinal Cord Hemodynamic Responses to Epidural Electrical Stimulation: A Feasibility Study.

This study presents the first implementation of functional ultrasound (fUS) imaging of the spinal cord to monitor local hemodynamic response to epidural electrical spinal cord stimulation (SCS) on two small and large animal models. SCS has been successfully applied to control chronic refractory pain and recently was evolved to alleviate motor impairment in Parkinson's disease and after spinal cord injury. At present, however, the mechanisms underlying SCS remain unclear, and current methods for monitoring SCS are limited in their capacity to provide the required sensitivity and spatiotemporal resolutions to evaluate functional changes in response to SCS. fUS is an emerging technology that has recently shown promising results in monitoring a variety of neural activities associated with the brain. Here we demonstrated the feasibility of performing fUS on two animal models during SCS. We showed in vivo spinal cord hemodynamic responses measured by fUS evoked by different SCS parameters. We also demonstrated that fUS has a higher sensitivity in monitoring spinal cord response than electromyography. The high spatial and temporal resolutions of fUS were demonstrated by localized measurements of hemodynamic responses at different spinal cord segments, and by reliable tracking of spinal cord responses to patterned electrical stimulations, respectively. Finally, we proposed optimized fUS imaging and post-processing methods for spinal cord. These results support feasibility of fUS imaging of the spinal cord and could pave the way for future systematic studies to investigate spinal cord functional organization and the mechanisms of spinal cord neuromodulation in vivo.

The Spinal Neurons Exhibit an ON-OFF and OFF-ON Firing Activity Around the Onset of Fictive Scratching Episodes in the Cat.

In a previous report, we found neurons with ON-OFF and OFF-ON firing activity in the obex reticular formation during scratching. The aim of the present study was to examine whether the spinal neurons also exhibit this type of activity in relation to the "postural stage" of fictive scratching in the cat. We found that the extensor and intermediate scratching neurons exhibit an ON-OFF firing rate; conversely, the flexor neurons show an OFF-ON activity, relative to every scratching episode. These patterns of spiking activity are similar to those found in neurons from the obex reticular formation during scratching. Our findings provide support to the following hypotheses. First, there is a possible functional link between supraspinal and spinal, ON-OFF and OFF-ON neuronal groups. Second, the fictive goal-directed motor action to maintain the fictive "postural stage" of the hindlimb during fictive scratching is associated with the neuronal tonic activity of the OFF-ON spinal neurons, whereas the ON-OFF spinal neurons are associated with an extensor tone that occurred prior the postural stage.

The Role of Functional Neuroanatomy of the Lumbar Spinal Cord in Effect of Epidural Stimulation.

In this study, the neuroanatomy of the swine lumbar spinal cord, particularly the spatial orientation of dorsal roots was correlated to the anatomical landmarks of the lumbar spine and to the magnitude of motor evoked potentials during epidural electrical stimulation (EES). We found that the proximity of the stimulating electrode to the dorsal roots entry zone across spinal segments was a critical factor to evoke higher peak-to-peak motor responses. Positioning the electrode close to the dorsal roots produced a significantly higher impact on motor evoked responses than rostro-caudal shift of electrode from segment to segment. Based on anatomical measurements of the lumbar spine and spinal cord, significant differences were found between L1-L4 to L5-L6 segments in terms of spinal cord gross anatomy, dorsal roots and spine landmarks. Linear regression analysis between intersegmental landmarks was performed and L2 intervertebral spinous process length was selected as the anatomical reference in order to correlate vertebral landmarks and the spinal cord structures. These findings present for the first time, the influence of spinal cord anatomy on the effects of epidural stimulation and the role of specific orientation of electrodes on the dorsal surface of the dura mater in relation to the dorsal roots. These results are critical to consider as spinal cord neuromodulation strategies continue to evolve and novel spinal interfaces translate into clinical practice.

Tonically Active α5GABAA Receptors Reduce Motoneuron Excitability and Decrease the Monosynaptic Reflex.

Motoneurons, the final common path of the Central Nervous System (CNS), are under a complex control of its excitability in order to precisely translate the interneuronal pattern of activity into skeletal muscle contraction and relaxation. To fulfill this relevant function, motoneurons are provided with a vast repertoire of receptors and channels, including the extrasynaptic GABAA receptors which have been poorly investigated. Here, we confirmed that extrasynaptic α5 subunit-containing GABAA receptors localize with choline acetyltransferase (ChAT) positive cells, suggesting that these receptors are expressed in turtle motoneurons as previously reported in rodents. In these cells, α5GABAA receptors are activated by ambient GABA, producing a tonic shunt that reduces motoneurons' membrane resistance and affects their action potential firing properties. In addition, α5GABAA receptors shunted the synaptic excitatory inputs depressing the monosynaptic reflex (MSR) induced by activation of primary afferents. Therefore, our results suggest that α5GABAA receptors may play a relevant physiological role in motor control.

Modeling zero-lag synchronization of dorsal horn neurons during the traveling of electrical waves in the cat spinal cord.

The first electrophysiological evidence of the phenomenon of traveling electrical waves produced by populations of interneurons within the spinal cord was reported by our interdisciplinary research group. Two interesting observations derive from this study: first, the negative spontaneous cord dorsum potentials (CDPs) that are superimposed on the propagating sinusoidal electrical waves are not correlated with any scratching phase; second, these CDPs do not propagate along the lumbosacral spinal segments, but they appear almost simultaneously at different spinal segments. The aim of this study was to provide experimental data and a mathematical model to explain the simultaneous occurrence of traveling waves and the zero-lag synchronization of some CDPs.

Control of flowering and storage organ formation in potato by FLOWERING LOCUS T.

Seasonal fluctuations in day length regulate important aspects of plant development such as the flowering transition or, in potato (Solanum tuberosum), the formation of tubers. Day length is sensed by the leaves, which produce a mobile signal transported to the shoot apex or underground stems to induce a flowering transition or, respectively, a tuberization transition. Work in Arabidopsis, tomato and rice (Oryza sativa) identified the mobile FLOWERING LOCUS T (FT) protein as a main component of the long-range 'florigen', or flowering hormone, signal. Here we show that expression of the Hd3a gene, the FT orthologue in rice, induces strict short-day potato types to tuberize in long days. Tuber induction is graft transmissible and the Hd3a-GFP protein is detected in the stolons of grafted plants, transport of the fusion protein thus correlating with tuber formation. We provide evidence showing that the potato floral and tuberization transitions are controlled by two different FT-like paralogues (StSP3D and StSP6A) that respond to independent environmental cues, and show that an autorelay mechanism involving CONSTANS modulates expression of the tuberization-control StSP6A gene.

An intersegmental neuronal architecture for spinal wave propagation under deletions.

Recent studies have established and characterized the propagation of traveling electrical waves along the cat spinal cord during scratching, but the neuronal architecture that allows for the persistence of such waves even during periods of absence of bursts of motoneuron activity (deletions) is still unclear. Here we address this problem both theoretically and experimentally. Specifically, we monitored during long lasting periods of time the global electrical activity of spinal neurons during scratching. We found clear deletions of unaltered cycle in extensor activity without associated deletions of the traveling spinal wave. Furthermore, we also found deletions with a perturbed cycle associated with a concomitant absence of the traveling spinal wave. Numerical simulations of an asymmetric two-layer model of a central-pattern generator distributed longitudinally along the spinal cord qualitatively reproduce the sinusoidal traveling waves, and are able to replicate both classes of deletions. We believe these findings shed light into the longitudinal organization of the central-pattern generator networks in the spinal cord.

Propagation of sinusoidal electrical waves along the spinal cord during a fictive motor task.

We present for the first time direct electrophysiological evidence of the phenomenon of traveling electrical waves produced by populations of interneurons within the spinal cord. We show that, during a fictive rhythmic motor task, scratching, an electrical field potential of spinal interneurons takes the shape of a sinuous wave, "sweeping" the lumbosacral spinal cord rostrocaudally with a mean speed of approximately 0.3 m/s. We observed that traveling waves and scratching have the same cycle duration and that duration of the flexor phase, but not of the extensor phase, is highly correlated with the cycle duration of the traveling waves. Furthermore, we found that the interneurons from the deep dorsal horn and the intermediate nucleus can generate the spinal traveling waves, even in the absence of motoneuronal activity. These findings show that the sinusoidal field potentials generated during fictive scratching could be a powerful tool to disclose the organization of central pattern generator networks.