Mechanisms of stepping rhythm formation in decerebrated and chronic spinal cats under epidural stimulation

A study was made of the stepping pattern formation in decerebrated and in chronic spinal cats during epidural stimulation (ES). The hindlimb stepping performance depended on the parameters of ES and afferent input. At non-optimal ES parameters, no stepping was induced, only muscle reflexes followed the stimulation rhythm. Optimized ES (3–5 Hz, 50–100 μA for decerebrated and 20–30 Hz, 150–250 μA for spinal cats) evoked coordinated stepping movements at a natural rate (0.8–1 Hz) accompanied by electromyographic burst activity of the corresponding muscles. In decerebrated cats, the bursts are formed owing to modulation of early responses and the late polysynaptic activity. In chronic spinal cats, this process is mainly due to amplitude modulation of the early responses. Formation of the stepping pattern in decerebrated cats involves spinal interneurons responsible for the polysynaptic activity, which allows its correction based on processing the afferent signals. Activation of this system in chronic spinal cats can be realized by afferent stimulation alone, without ES.     

The role of skin afferent in regulation of locomotion evoked by electrical epidural stimulation of spinal cord in decerebrated cats

The role of hindpaw skin afferent input in the locomotor pattern formation induced by epidural spinal cord stimulation was investigated in decerebrated cats. Locomotor activity was evoked by continuous 3-5Hz stimulation of dorsal surface of L4-L5 spinal segments. Kinematic and electromyographic activity (EMG) of m. Quadriceps, m. Semitendinosus, m. Tibialis anterior an m. Gastrocnemius lateralis before and after blocking of skin receptors in one hind limb were recorded. In addition, reflex responses in the hind limb muscles to epidural stimulation with frequency 0.5 Hz were analysed. Blocking of skin receptors of the foot with chlorothane paw irrigation or 2 % lidocaine administrated into the hind paw was performed. After blocking of skin receptors of the foot the stepping pattern changed. Stepping with dorsal foot placement and dragging during swing phase was observed. Duration of stance phase significantly decreased. Inhibition of polysynaptic activity of proximal and distal extensor muscles and distal flexor muscles of hind paw during locomotion was found. Monosynaptic responses after blocking of skin receptors of the foot changed insignificantly.     

Significance of peripheral feedback in stepping movement generation under epideral spinal cord stimulation

In acute experiments on decerebrated and spinalized cats, the role of peripheral afferent input from hindlimbs in stepping patterns formation under epidural spinal cord stimulation (ESCS), was investigated. The hindlimb muscles' electromyographic activity and kinematic parameters of evoked stepping were analyzed. It has been shown that epidural stimulation (20-100 microA, 5 Hz) of L4-L5 spine segments induced coordinated stepping on the treadmill belt. In conditions of weight-bearing support (stopped treadmill, hindlimbs lifted above the treadmill), the stepping rhythmic was unstable, stepping cycle period and its internal structure having changed as well. With increased speed of locomotion the stepping frequency increased due to the duration of the support phase decreasing. Forward stepping could be reversed to backward stepping by changing the direction of the treadmill belt movement. In 2-4 hours after complete spinal transection (T8-T9), the epidural stimulation elicited stepping movements on a moving treadmill only. It was found that the influence of peripheral feedback on initiation of the stepping after spinalization increased. Peripheral feedback seems to play a major role in determining the fundamental features of motor output during the ESCS.     

Locomotion induced by epidural stimulation in a decerebrated cat after spinal cord injury

The effect of partial and complete spinal cord transection (Th7–Th8) on locomotor activity evoked in decerebrated cats by electrical epidural stimulation (segment L5, 80–100 μA, 0.5 ms at 5 Hz) has been investigated. Transection of dorsal columns did not substantially influence the locomotion. Disruption of the ventral spinal quadrant resulted in deterioration and instability of the locomotor rhythm. Injury to lateral or medial descending motor systems led to redistribution of the tone in antagonist muscles. Locomotion could be evoked by epidural stimulation within 20 h after complete transection of the spinal cord. The restoration of polysynaptic components in EMG responses correlated with recovery of the stepping function. The data obtained confirm that initiation of locomotion under epidural stimulation is caused by direct action on intraspinal systems responsible for locomotor regulation. With intact or partially injured spinal cord, this effect is under the influence of supraspinal motor systems correcting and stabilizing the evoked locomotor pattern.     

Control of the human and animal locomotor activity in the absence of supraspinal effects

In patients deprived of supraspinal effects, electrical epidural stimulation of the spinal cord's dorsal surface at the level of 2nd lumbar segment induces step-like movements accompanied by respective electromyographic activity of the leg's muscles. Triggering of the step-like movements occurs at certain parameters of the stimulation. The data obtained suggest that human spinal cord has networks of interneurons-generators of the step-like movements. A leading role of the spinal cord's propriospinal system in activation of spinal generators of stepping under epidural influences was shown in cats.     

Initiation of locomotor activity in spinalized cats by epidural stimulation of the spinal cord]

An area initiating locomotor activity under conditions of the spinal cord surface stimulation (SCS) was found at the upper border of the lumbar enlargement (L4-5 spinal segments). Parameters of the SCS for activation of the spinal stepping generator (SSG) were identified. Activation of the SSG under the SCS was shown to occur because of interaction of the spinal and peripheral mechanisms. Primary role in the SSG initiation during the SCS belongs to the propriospinal system of the dorsolateral funiculi.     

Convergence of forelimb afferent actions on C7-Th1 propriospinal neurones bilaterally projecting to sacral segments of the cat spinal cord

Propriospinal neurones located in the cervical enlargement and projecting bilaterally to sacral segments of the spinal cord were investigated electrophysiologically in eleven deeply anaesthetized cats. Excitatory or inhibitory postsynaptic potentials from forelimb afferents were recorded following stimulation of deep radial (DR), superficial radial (SR), median (Med) and ulnar (Uln) nerves. 26 cells were recorded from C7, 22 from C8 and 3 from Th1 segments. The majority of the cells were located in the Rexed's laminae VIII and the medial part of the lamina VII. In 10 cases no afferent input from the forelimb afferents was found. In the remaining neurones effects were evoked mostly from DR (88%) and Med (63%), less often from SR (46%) and Uln (46%). Inhibitory actions were more frequent than excitatory. The highest number of IPSPs was evoked from high threshold flexor reflex afferents (FRA)--all connections were polysynaptic. However, inhibitory actions were often evoked from group I or II muscle afferents (polysynaptic or disynaptic) and, less frequently, from cutaneous afferents (mostly polysynaptic). Di- or polysynaptic IPSPs often accompanied monosynaptic EPSPs from group I or II muscle afferents. Disynaptic or polysynaptic EPSPs from muscle and cutaneous afferents were also recorded in many neurones, while polysynaptic EPSPs from FRA were observed only exceptionally. Various patterns of convergence in individual neuronal subpopulations indicate that they integrate different types of the afferent input from various muscle and cutaneous receptors of the distal forelimb. They transmit this information to motor centers controlling hind limb muscles, forming a part of the system contributing to the process of coordination of movements of fore--and hind--limbs.     

The heterogeneity of the excitatory synaptic inputs in the spinal motor neurons of the frog Rana ridibunda

The synaptic responses induced in motoneurones by the stimulations of the dorsal root (DR), single afferent fibres and reticular formation (RF) were intracellularly recorded in the isolated frog spinal cord. It was shown that argiopine (the selective blocker of glutamate receptors of non-NMDA type) in concentrations ranging from 3.10(-7) to 1.10(-5) M effectively suppressed the di- and polysynaptic, but not the monosynaptic components of EPSP's induced by DR stimulation. The initial reaction to argiopine consisted of the increase of this monosynaptic component of EPSP. In the same concentrations range, argiopine reduced both mono- and polysynaptic EPSP, evoked by RF stimulation. 2-amino-phosphonovaleric acid (1.10(-4) M) did not affect, whereas the kinurenate (1--2.10(-3) M) completely blocked the amplitude of all kinds of synaptic responses. The various effects of argiopine on the responses induced by microstimulation of presynaptic nerve terminals were observed. The data obtained speak in favour of heterogeneity of monosynaptic excitatory inputs in the motoneurones of frog spinal cord. Being the glutamatergic by nature, the inputs differ in the properties of postsynaptic receptors. All of these receptors concerning to non NMDA-type can be divided to argiopine-sensitive and argiopine-resistant. The first seem to be involved in the monosynaptic connections of RF and the second--in those of primary afferents with motoneurones.     

Sensorimotor regulation of movements: Novel strategies for the recovery of mobility

A series of observations have provided important insight into properties of the spinal as well as supraspinal circuitries that control posture and movement. We have demonstrated that spinal rats can regain full weight-bearing standing and stepping over a range of speeds and directions with the aid of electrically enabling motor control (eEmc), pharmacological modulation (fEmc), and training [1, 2]. Also, we have reported that voluntary control movements of individual joints and limbs can be regained after complete paralysis in humans [3, 4]. However, the ability to generate significant levels of voluntary weight-bearing stepping with or without epidural spinal cord stimulation remains limited. Herein we introduce a novel method of painless transcutaneous electrical enabling motor control (pcEmc) and sensory enabling motor control (sEmc) strategy to neuromodulate the physiological state of the spinal cord. We have found that a combination of a novel non-invasive transcutaneous spinal cord stimulation and sensory-motor stimulation of leg mechanoreceptors can modulate the spinal locomotor circuitry to state that enables voluntary rhythmic locomotor movements.     

Synaptic processes in thoracic α-motoneurons evoked by segmental afferent stimulation

Synaptic processes in various functional groups of thoracic motoneurons (Th₉-Th₁₁) evoked by stimulation of segmental nerves were investigated in anesthetized and decerebrate cats. No reciprocal relations were found between these groups of motoneurons. Only excitatory mono- and polysynaptic responses were recorded in the motoneurons of the principal intercostal nerve following stimulation of the homonymous nerve. Activation of the afferents of the external intercostal muscle and dorsal branches does not cause noticeable synaptic processes in these motoneurons; much more rarely it is accompanied by the development of low-amplitude polysynaptic EPSP's. In motoneurons of the dorsal branches, stimulation of homonymous nerves leads to the appearance of simple, short-latent EPSP's. Late responses of the IPSP or EPSP - IPSP type with a predominance of the inhibitory component were observed in most motoneurons of this type following activation of the afferent fibers of the principal intercostal nerve. In other motoneurons of the dorsal muscles, stimulation of the main intercostal nerve (and nerve of the external intercostal muscle) did not evoke apparent synpatic processes. EPSP's (mono- and polysynaptic) appeared in the motoneurons of the external intercostal muscle following stimulation of the homonymous and main intercostal nerves. Activation of the afferents of the dorsal branches was ineffective. The character of the synaptic responses of the respiratory motoneurons to segmental afferent stimulation, investigated under conditions of spontaneous respiration, was different. The characteristics of synaptic activation of thoracic motoneurons by segmental afferents under conditions of hyperventilation apnea and during spontaneous breathing of the animals are discussed.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 2, No. 3, pp. 279–288, May–June, 1970.     

The influence of vibration on spinal alpha-motoneurons excitability in static conditions and during evoked stepping in human

In healthy human the excitability of spinal alpha-motoneurons under application of vibrostimulation (20-60 Hz) to different leg muscles was investigated both in stationary condition and during stepping movements caused by vibration in the condition of suspended leg. In 15 subjects the amplitude of H-reflex were compared under vibration of rectus femoris (RF) and biceps femoris (BF) muscles of left leg as well during vibration of rectus femoris of contralateral, motionless leg in three spatial positions: upright, supine and on right side of body with suspended left leg. In dynamic conditions the amount of H-reflex was compared during evoked and voluntary stepping at 8 intervals of step cycle. In all body positions the vibration of each ipsilateral leg muscles caused significant suppression of H-reflex, this suppression was more prominent in the air-stepping conditions. The vibration of contralateral leg RF muscle had a weak influence on the amplitude of H-reflex. In 7 subjects the muscle vibration of ipsilateral and contralateral legs generated stepping movements. During evoked "air-stepping" H-reflex had different amplitudes in different phases of step cycle. At the same time the differences between responses under voluntary and non-voluntary stepping were revealed only in stance phase. Thus, different degree of H-reflex suppression by vibration under different body position in space depends on, it seems to be, from summary afferent inflows to spinal cord interneurons, which participate in regulation of posture and locomotion. Seemingly, the increasing of spinal cord neurons excitability occurs under involuntary air-stepping in swing phase, which is necessary for activation of locomotor automatism under unloading leg conditions.     

Effect of hypoxia on the excitability of two cranial reflexes in unanaesthetized fetal sheep

In fetal sheep acute hypoxia causes a decreased incidence of breathing movements and motor activity, and the excitability of polysynaptic reflexes in the hindlimbs is depressed. To determine whether this inhibitory effect extends to other areas in the fetal CNS, we have studied the effect of hypoxia on two reflexes with cranial pathways. The digastric (jaw opening) reflex was elicited by stimulation of the dental nerve through a pair of stainless steel electrodes implanted into the mandible (4 fetuses). The thyroarytenoid muscle of the larynx was reflexly activated by stimulation of the superior laryngeal nerve by a cuff electrode (4 fetuses). Low level stimulation at 1.5-2 X threshold was repeated at approximately 2 min intervals for 3-4 h; the stimulation did not alter the pattern of electrocortical activity, breathing movements, or cause arousal. The amplitude of the digastric reflex was greatest during low voltage electrocortical activity; conversely, the amplitude of the thyroarytenoid reflex was greatest during high voltage electrocortical activity. Isocapnic hypoxia lasting 30-60 min (16 trials), in which the PaO2 was reduced to 12-14 mmHg, did not reduce the amplitude of either reflex. The reduction of thyroarytenoid reflex amplitude which normally occurred during low voltage electrocortical activity was not present during hypoxia. These experiments show that the inhibitory effects of hypoxia on spinal reflexes, breathing movements and motor activity do not include these cranial pathways.     

Response of external inspiration to the movements induced by transcutaneous spinal cord stimulation

The dynamic of the parameters of lung ventilation and gas exchange have been studied in 10 young male subjects during involuntary stepping movements induced by transcutaneous spinal cord electrical stimulation applied in the projection of T ₁₁–T ₁₂ vertebrae and during voluntary stepping movements. It has been found that the transcutaneous spinal cord stimulation inducing stepping movements leads to an increase in breathing frequency and a reduction in tidal volume. These effects may be mediated by some neurogenic factors associated with muscular activity during stepping movements, the activation of abdominal expiratory muscles, and the interaction between the stepping pattern and breathing generators.     

Transcutaneous electrical stimulation of the spinal cord: non-invasive tool for activation of locomotor circuitry in human

A new tool for locomotor circuitry activation in the non-injured human by transcutaneous electrical spinal cord stimulation (tSCS) has been described. We show that continuous tSCS over T11-T12 vertebrae at 5-40 Hz induced involuntary locomotor-like stepping movements in subjects with their legs in a gravity-independent position. The increase of frequency of tSCS from 5 to 30 Hz augmented the amplitude of evoked stepping movements. The duration of cycle period did not depend on frequency of tSCS. During tSCS the hip, knee and ankle joints were involved in the stepping performance. It has been suggested that tSCS activates the locomotor circuitry through the dorsal roots. It appears that tSCS can be used as a non-invasive method in rehabilitation of spinal pathology.     

Spinal cord evoked potentials and edema in the pathophysiology of rat spinal cord injury

Summary The possibility that nitric oxide is somehow involved in the early bioelectrical disturbances following spinal cord injury in relation to the later pathophysiology of the spinal cord was examined in a rat model of spinal cord trauma. A focal trauma to the rat spinal cord was produced by an incision of the right dorsal horn of the T 10–11 segments under urethane anaesthesia. The spinal cord evoked potentials (SCEP) were recorded using epidural electrodes placed over the T9 and T12 segments of the cord following supramaximal stimulation of the right tibial and sural nerves in the hind leg. Trauma to the spinal cord significantly attenuated the SCEP amplitude (about 60%) immediately after injury which persisted up to 1h. However, a significant increase in SCEP latency was seen at the end of 5h after trauma. These spinal cord segments exhibited profound upregulation of neuronal nitric oxide synthase (NOS) immunoreactivity, and the development of edema and cell injury. Pretreatment with a serotonin synthesis inhibitor drug p-chlorophenylalanine (p-CPA) or an anxiolytic drug diazepam significantly attenuated the decrease in SCEP amplitude, upregulation of NOS, edema and cell injury. On the other hand, no significant reduction in SCEP amplitude, NOS immunolabelling, edema or cell changes were seen after injury in rats pretreated with L-NAME. These observations suggest that nitric oxide is somehow involved in the early disturbances of SCEP and contribute to the later pathophysiology of spinal cord injury.     

The effect of L-DOPA on potentials of the dorsal surface of the spinal cord evoked by the stimulation of primary afferents in newborn rat pups

In experiments on 5-30-day rat puppies, studies have been made of the effect of L-DOPA (100 mg/kg, intraperitoneally) on the activity of interneurones of the dorsal horn of the spinal cord as revealed from the parameters of potentials of the dorsal surface of the spinal cord. The specific pattern of reaction in 5-day animals is manifested in a succession of inhibitory inhibition and increase in the activity of neurones monosynaptically activated by low-threshold afferents. Both the amplitude and duration of polysynaptic components of the potentials of the dorsal surface are rather high. From the 7th day, deep and stable inhibition is observed which is accompanied by a decrease in the amplitude of all components of the potential of the dorsal surface. At later stages of ontogenesis, a decrease is observed in the inhibitory effect of L-DOPA on the activity of interneurones which monosynaptically contact with low- and especially high-threshold afferents; in contrast to earlier stages, but similar to adult animals, evident inhibition was revealed in the activity of interneurones which have polysynaptic contacts with high-threshold afferents and afferents of flexor reflex. Thus, within the first weeks of postnatal life, basic qualitative changes are observed in the pattern of the reaction of interneurones of the dorsal horn to exogenic catecholamines.     

Neurons of upper cervical segments responding to stimulation of the bulbar locomotor strip

Synaptic responses of single neurons to stimulation of the bulbar "locomotor strip" were recorded extracellularly from superior cervical segments in mesencephalic cats. With a strength of stimulation of about 30 µA these responses usually had a latent period of 2–7 msec and they arose in neurons located at a depth of between 2 and 4 mm from the dorsal surface (Rexed's laminae V–VIII). These neurons could not be excited antidromically by stimulation of the lumbar or lower cervical segments. However, antidromic responses could be evoked by stimulation of a region located 3–5 mm caudally to the site of recording. It is suggested that neurons of segments C2 and C3 excited by stimulation of the locomotor strip are components of a cell column along which activity spreads polysynaptically in the direction of spinal stepping generators.Institute for Problems in Information Transmission, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 11, No. 3, pp. 245–253, May–June, 1979.     

Synaptic activation of thoracic spinal interneurons by reticulospinal pathways

Synaptic processes of 119 thoracic spinal interneurons (T10–11) were investigated in anesthetized cats in response to stimulation of the medial and central zones of the gigantocellular nucleus in the medulla and the ventral columns of the spinal cord. Fast (90–130 m/sec) reticulospinal fibers running in the ventral column were found to produce monosynaptic or disynaptic excitation of interneurons of Rexed's layers VII–VIII, which are connected monosynaptically with group I muscle afferents, and interneurons excited both by group I muscle afferents and low-threshold cutaneous afferents. In most neurons of layer IV, connected monosynaptically with low-threshold cutaneous afferents, and in neurons of layers VII–VIII excited by afferents of the flexor reflex no marked postsynaptic processes were observed during stimulation of the reticular formation. Excitatory, inhibitory, and mixed PS Ps during activation of reticulospinal fibers were found in 14 neurons, high-threshold afferents in which evoked predominantly polysynaptic IPSPs. Seventeen neurons activated monosynaptically by reticulospinal fibers and not responding to stimulation of segmental afferents were found in the medial part of the ventral horn (layers VII–VIII).A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 4, No. 6, pp. 566–578, November–December, 1972.