Neuromechanical Strategies for Obstacle Negotiation during Overground Locomotion following Incomplete Spinal Cord Injury in Adult Cats.

Following incomplete spinal cord injury in animals, including humans, substantial locomotor recovery can occur. However, functional aspects of locomotion, such as negotiating obstacles, remains challenging. We collected kinematic and electromyography data in 10 adult cats (5 males, 5 females) before and at weeks 1-2 and 7-8 after a lateral mid-thoracic hemisection on the right side of the cord while they negotiated obstacles of three different heights. Intact cats always cleared obstacles without contact. At weeks 1-2 after hemisection, the ipsilesional right hindlimb contacted obstacles in ∼50% of trials, triggering a stumbling corrective reaction or absent responses, which we termed Other. When complete clearance occurred, we observed exaggerated ipsilesional hindlimb flexion when crossing the obstacle with contralesional Left limbs leading. At weeks 7-8 after hemisection, the proportion of complete clearance increased, Other responses decreased, and stumbling corrective reactions remained relatively unchanged. We found redistribution of weight support after hemisection, with reduced diagonal supports and increased homolateral supports, particularly on the left contralesional side. The main neural strategy for complete clearance in intact cats consisted of increased knee flexor activation. After hemisection, ipsilesional knee flexor activation remained, but it was insufficient or more variable as the limb approached the obstacle. Intact cats also increased their speed when stepping over an obstacle, an increase that disappeared after hemisection. The increase in complete clearance over time after hemisection paralleled the recovery of muscle activation patterns or new strategies. Our results suggest partial recovery of anticipatory control through neuroplastic changes in the locomotor control system.SIGNIFICANCE STATEMENT Most spinal cord injuries (SCIs) are incomplete and people can recover some walking functions. However, the main challenge for people with SCIs that do recover a high level of function is to produce a gait that can adjust to everyday occurrences, such as turning, stepping over an obstacle, etc. Here, we use the cat model to answer two basic questions: How does an animal negotiate an obstacle after an incomplete SCI and why does it fail to safely clear it? We show that the inability to clear an obstacle is because of improper activation of muscles that flex the knee. Animals recover a certain amount of function thanks to new strategies and changes within the nervous system.

Simultaneous control of forward and backward locomotion by spinal sensorimotor circuits.

Mammals walk in different directions, such as forward and backward. In human infants/adults and decerebrate cats, one leg can walk forward and the other backward simultaneously on a split-belt treadmill, termed hybrid or bidirectional locomotion. The purpose of the present study was to determine if spinal sensorimotor circuits generate hybrid locomotion and if so, how the limbs remain coordinated. We tested hybrid locomotion in 11 intact cats and in five following complete spinal thoracic transection (spinal cats) at three treadmill speeds with the hindlimbs moving forward, backward or bidirectionally. All intact cats generated hybrid locomotion with the forelimbs on a stationary platform. Four of five spinal cats generated hybrid locomotion, also with the forelimbs on a stationary platform, but required perineal stimulation. During hybrid locomotion, intact and spinal cats positioned their forward and backward moving hindlimbs caudal and rostral to the hip, respectively. The hindlimbs maintained consistent left-right out-of-phase alternation in the different stepping directions. Our results suggest that spinal locomotor networks generate hybrid locomotion by following certain rules at phase transitions. We also found that stance duration determined cycle duration in the different locomotor directions/conditions, consistent with a common rhythm-generating mechanism for different locomotor directions. Our findings provide additional insight on how left-right spinal networks and sensory feedback from the limbs interact to coordinate the hindlimbs and provide stability during locomotion in different directions. KEY POINTS: Terrestrial mammals can walk forward and backward, which is controlled in part by spinal sensorimotor circuits. Humans and cats also perform bidirectional or hybrid locomotion on a split-belt treadmill with one leg going forward and the other going backward. We show that cats with a spinal transection can perform hybrid locomotion and maintain left-right out-of-phase coordination, indicating that spinal sensorimotor circuits can perform simultaneous forward and backward locomotion. We also show that the regulation of cycle duration and phase duration is conserved across stepping direction, consistent with a common rhythm-generating mechanism for different stepping directions. The results help us better understand how spinal networks controlling the left and right legs enable locomotion in different directions.

Changes in intra- and interlimb reflexes from hindlimb cutaneous afferents after staggered thoracic lateral hemisections during locomotion in cats.

When the foot dorsum contacts an obstacle during locomotion, cutaneous afferents signal central circuits to coordinate muscle activity in the four limbs. Spinal cord injury disrupts these interactions, impairing balance and interlimb coordination. We evoked cutaneous reflexes by electrically stimulating left and right superficial peroneal nerves before and after two thoracic lateral hemisections placed on opposite sides of the cord at 9- to 13-week interval in seven adult cats (4 males and 3 females). We recorded reflex responses in ten hindlimb and five forelimb muscles bilaterally. After the first (right T5-T6) and second (left T10-T11) hemisections, coordination of the fore- and hindlimbs was altered and/or became less consistent. After the second hemisection, cats required balance assistance to perform quadrupedal locomotion. Short-latency reflex responses in homonymous and crossed hindlimb muscles largely remained unaffected after staggered hemisections. However, mid- and long-latency homonymous and crossed responses in both hindlimbs occurred less frequently after staggered hemisections. In forelimb muscles, homolateral and diagonal mid- and long-latency response occurrence significantly decreased after the first and second hemisections. In all four limbs, however, when present, short-, mid- and long-latency responses maintained their phase-dependent modulation. We also observed reduced durations of short-latency inhibitory homonymous responses in left hindlimb extensors early after the first hemisection and delayed short-latency responses in the right ipsilesional hindlimb after the first hemisection. Therefore, changes in cutaneous reflex responses correlated with impaired balance/stability and interlimb coordination during locomotion after spinal cord injury. Restoring reflex transmission could be used as a biomarker to facilitate locomotor recovery. KEY POINTS: Cutaneous afferent inputs coordinate muscle activity in the four limbs during locomotion when the foot dorsum contacts an obstacle. Thoracic spinal cord injury disrupts communication between spinal locomotor centres located at cervical and lumbar levels, impairing balance and limb coordination. We investigated cutaneous reflexes during quadrupedal locomotion by electrically stimulating the superficial peroneal nerve bilaterally, before and after staggered lateral thoracic hemisections of the spinal cord in cats. We showed a loss/reduction of mid- and long-latency responses in all four limbs after staggered hemisections, which correlated with altered coordination of the fore- and hindlimbs and impaired balance. Targeting cutaneous reflex pathways projecting to the four limbs could help develop therapeutic approaches aimed at restoring transmission in ascending and descending spinal pathways.

Forelimb movements contribute to hindlimb cutaneous reflexes during locomotion in cats.

During quadrupedal locomotion, interactions between spinal and supraspinal circuits and somatosensory feedback coordinate forelimb and hindlimb movements. How this is achieved is not clear. To determine whether forelimb movements modulate hindlimb cutaneous reflexes involved in responding to an external perturbation, we stimulated the superficial peroneal nerve in six intact cats during quadrupedal locomotion and during hindlimb-only locomotion (with forelimbs standing on stationary platform) and in two cats with a low spinal transection (T12-T13) during hindlimb-only locomotion. We compared cutaneous reflexes evoked in six ipsilateral and four contralateral hindlimb muscles. Results showed similar occurrence and phase-dependent modulation of short-latency inhibitory and excitatory responses during quadrupedal and hindlimb-only locomotion in intact cats. However, the depth of modulation was reduced in the ipsilateral semitendinosus during hindlimb-only locomotion. Additionally, longer-latency responses occurred less frequently in extensor muscles bilaterally during hindlimb-only locomotion, whereas short-latency inhibitory and longer-latency excitatory responses occurred more frequently in the ipsilateral and contralateral sartorius anterior, respectively. After spinal transection, short-latency inhibitory and excitatory responses were similar to both intact conditions, whereas mid- or longer-latency excitatory responses were reduced or abolished. Our results in intact cats and the comparison with spinal-transected cats suggest that the absence of forelimb movements suppresses inputs from supraspinal structures and/or cervical cord that normally contribute to longer-latency reflex responses in hindlimb extensor muscles.NEW & NOTEWORTHY During quadrupedal locomotion, the coordination of forelimb and hindlimb movements involves central circuits and somatosensory feedback. To demonstrate how forelimb movement affects hindlimb cutaneous reflexes during locomotion, we stimulated the superficial peroneal nerve in intact cats during quadrupedal and hindlimb-only locomotion as well as in spinal-transected cats during hindlimb-only locomotion. We show that forelimb movement influences the modulation of hindlimb cutaneous reflexes, particularly the occurrence of long-latency reflex responses.

The Spinal Control of Backward Locomotion.

Animal locomotion requires changing direction, from forward to backward. Here, we tested the hypothesis that sensorimotor circuits within the spinal cord generate backward locomotion and adjust it to task demands. We collected kinematic and electromyography (EMG) data during forward and backward locomotion at different treadmill speeds before and after complete spinal transection in six adult cats (three males and three females). After spinal transection, five/six cats performed backward locomotion, which required tonic somatosensory input in the form of perineal stimulation. One spinal cat performed forward locomotion but not backward locomotion while two others stepped backward but not forward. Spatiotemporal adjustments to increasing speed were similar in intact and spinal cats during backward locomotion and strategies were similar to forward locomotion, with shorter cycle and stance durations and longer stride lengths. Patterns of muscle activations, including muscle synergies, were similar for forward and backward locomotion in spinal cats. Indeed, we identified five muscle synergies that were similar during forward and backward locomotion. Lastly, spinal cats also stepped backward on a split-belt treadmill, with the left and right hindlimbs stepping at different speeds. Therefore, our results show that spinal sensorimotor circuits generate backward locomotion but require additional excitability compared with forward locomotion. Similar strategies for speed modulation and similar patterns of muscle activations and muscle synergies during forward and backward locomotion are consistent with a shared spinal locomotor network, with sensory feedback from the limbs controlling the direction.SIGNIFICANCE STATEMENT Animal locomotion requires changing direction, including forward, sideways and backward. This paper shows that the center controlling locomotion within the spinal cord can produce a backward pattern when instructed by sensory signals from the limbs. However, the spinal locomotor network requires greater excitability to produce backward locomotion compared with forward locomotion. The paper also shows that the spinal network controlling locomotion in the forward direction also controls locomotion in the backward direction.

Modulation of the gait pattern during split-belt locomotion after lateral spinal cord hemisection in adult cats.

Most previous studies investigated the recovery of locomotion in animals and people with incomplete spinal cord injury (SCI) during relatively simple tasks (e.g., walking in a straight line on a horizontal surface or a treadmill). We know less about the recovery of locomotion after incomplete SCI in left-right asymmetric conditions, such as turning or stepping along circular trajectories. To investigate this, we collected kinematic and electromyography data during split-belt locomotion at different left-right speed differences before and after a right thoracic lateral spinal cord hemisection in nine adult cats. After hemisection, although cats still performed split-belt locomotion, we observed several changes in the gait pattern compared with the intact state at early (1-2 wk) and late (7-8 wk) time points. Cats with larger lesions showed new coordination patterns between the fore- and hindlimbs, with the forelimbs taking more steps. Despite this change in fore-hind coordination, cats maintained consistent phasing between the fore- and hindlimbs. Adjustments in cycle and phase (stance and swing) durations between the slow and fast sides allowed animals to maintain 1:1 left-right coordination. Periods of triple support involving the right (ipsilesional) hindlimb decreased in favor of quad support and triple support involving the other limbs. Step and stride lengths decreased with concurrent changes in the right fore- and hindlimbs, possibly to avoid interference. The above adjustments in the gait pattern allowed cats to retain the ability to locomote in asymmetric conditions after incomplete SCI. We discuss potential plastic neuromechanical mechanisms involved in locomotor recovery in these conditions.NEW & NOTEWORTHY Everyday locomotion often involves left-right asymmetries, when turning, walking along circular paths, stepping on uneven terrains, etc. To show how incomplete spinal cord injury affects locomotor control in asymmetric conditions, we collected data before and after a thoracic lateral spinal hemisection on a split-belt treadmill with one side stepping faster than the other. We show that adjustments in kinematics and muscle activity allowed cats to retain the ability to perform asymmetric locomotion after hemisection.

Cutaneous inputs from perineal region facilitate spinal locomotor activity and modulate cutaneous reflexes from the foot in spinal cats.

It is well known that mechanically stimulating the perineal region potently facilitates hindlimb locomotion and weight support in mammals with a spinal transection (spinal mammals). However, how perineal stimulation mediates this excitatory effect is poorly understood. We evaluated the effect of mechanically stimulating (vibration or pinch) the perineal region on ipsilateral (9-14 ms onset) and contralateral (14-18 ms onset) short-latency cutaneous reflex responses evoked by electrically stimulating the superficial peroneal or distal tibial nerve in seven adult spinal cats where hindlimb movement was restrained. Cutaneous reflexes were evoked before, during, and after mechanical stimulation of the perineal region. We found that vibration or pinch of the perineal region effectively triggered rhythmic activity, ipsilateral and contralateral to nerve stimulation. When electrically stimulating nerves, adding perineal stimulation modulated rhythmic activity by decreasing cycle and burst durations and by increasing the amplitude of flexors and extensors. Perineal stimulation also disrupted the timing of the ipsilateral rhythm, which had been entrained by nerve stimulation. Mechanically stimulating the perineal region decreased ipsilateral and contralateral short-latency reflex responses evoked by cutaneous inputs, a phenomenon we observed in muscles crossing different joints and located in different limbs. The results suggest that the excitatory effect of perineal stimulation on locomotion and weight support is mediated by increasing the excitability of central pattern-generating circuitry and not by increasing excitatory inputs from cutaneous afferents of the foot. Our results are consistent with a state-dependent modulation of reflexes by spinal interneuronal circuits.

Mechanically stimulating the lumbar region inhibits locomotor-like activity and increases the gain of cutaneous reflexes from the paws in spinal cats.

Mechanically stimulating the dorsal lumbar region inhibits locomotion and reduces weight support during standing in rabbits and cats. However, how this inhibitory effect from the lumbar skin is mediated is poorly understood. Here we evaluated the effect of mechanically stimulating (vibration or pinch) the dorsal lumbar region on short-latency (8- to 13-ms onset) cutaneous reflex responses, evoked by electrically stimulating the superficial peroneal or distal tibial nerves, in seven adult cats with a low thoracic spinal transection (spinal cats). Cutaneous reflexes were evoked before, during, and after mechanical stimulation of the dorsal lumbar region. We found that mechanically stimulating the lumbar region by vibration or manual pinch abolished alternating bursts of activity between flexors and extensors initiated by nerve stimulation. The activity of extensor muscles was abolished bilaterally, whereas the activity of some ipsilateral flexor muscles was sustained during vibration/pinch. Mechanically stimulating the lumbar region increased ipsilateral and contralateral short-latency excitatory responses evoked by cutaneous inputs, a phenomenon that was generalized to muscles crossing different joints and located in different limbs. Our results indicate that the inhibitory effect on locomotion and weight support is not mediated by reducing cutaneous reflex gain and instead points to an inhibition of central pattern-generating circuitry, particularly the extensor component. The results provide greater insight into interactions between different types of somatosensory inputs within spinal motor circuits.NEW & NOTEWORTHY Vibration or pinch of the lumbar region in spinal-transected cats abolished alternating bursts of activity between flexors and extensors initiated by nerve stimulation. Mechanically stimulating the lumbar region increased ipsilateral and contralateral short-latency excitatory responses evoked by cutaneous inputs in hindlimb muscles. Sensory inputs from mechanoreceptors of the lumbar region do not mediate their inhibitory effect on locomotion and weight support by reducing the gain of short-latency excitatory cutaneous reflexes from the foot.

A Spinal Mechanism Related to Left-Right Symmetry Reduces Cutaneous Reflex Modulation Independently of Speed During Split-Belt Locomotion.

Task- and phase-dependent reflex modulation during locomotion is well established, but we do not know the signals driving this modulation. To determine whether signals related to left-right symmetry of the locomotor pattern modulate cutaneous reflexes, we stimulated the superficial peroneal nerve in five intact female cats and in four spinal-transected cats (spinal cats, two males and two females) during split-belt locomotion at different left-right speeds. We compared cutaneous reflexes evoked in three ipsilateral and two contralateral hindlimb muscles during split-belt locomotion with those evoked during tied-belt (equal left-right speeds) locomotion at matched speeds of the slow and fast limbs. Our results showed similar phase-dependent modulation of cutaneous reflexes during tied-belt and split-belt locomotion in intact and spinal cats. During tied-belt locomotion in intact cats, an increase in speed significantly increased reflex modulation from minimum to maximum values, whereas in spinal cats, we observed a significant decrease. However, in all muscles of intact and spinal cats, split-belt locomotion significantly reduced reflex modulation compared with tied-belt locomotion independently of which limb was stepping on the slow or fast belt. Additionally, reflex modulation correlated more with spatial left-right symmetry, as opposed to a temporal one, in intact and spinal cats. Our results indicate that signals related to left-right symmetry reduce cutaneous reflex modulation independently of speed via a spinal mechanism. We propose that asymmetric sensory feedback from the left and right legs alters the state of the spinal network, thereby reducing cutaneous reflexes to prevent inputs from destabilizing a potentially unstable pattern.SIGNIFICANCE STATEMENT When we contact an obstacle during walking, receptors in the skin send signals to the CNS to alter the trajectory of the leg to maintain balance. This response, or reflex, is different when the leg is in the air and when it is contacting the ground. The reflex also differs when we walk at different speeds. Here, we investigated this reflex when the left and right legs were walking at different speeds on a split-belt treadmill in cats. We show that the reflex is smaller during split-belt locomotion compared with when both legs are walking at equal speeds. We propose that the spinal locomotor network controlling walking reduces the reflex response to optimize balance when gait is unstable.

Intralimb and Interlimb Cutaneous Reflexes during Locomotion in the Intact Cat.

When the foot contacts an obstacle during locomotion, cutaneous inputs activate spinal circuits to ensure dynamic balance and forward progression. In quadrupeds, this requires coordinated reflex responses between the four limbs. Here, we investigated the patterns and phasic modulation of cutaneous reflexes in forelimb and hindlimb muscles evoked by inputs from all four limbs. Five female cats were implanted to record muscle activity and to stimulate the superficial peroneal and superficial radial nerves during locomotion. Stimulating these nerves evoked short-, mid-, and longer-latency excitatory and/or inhibitory responses in all four limbs that were phase-dependent. The largest responses were generally observed during the peak activity of the muscle. Cutaneous reflexes during mid-swing were consistent with flexion of the homonymous limb and accompanied by modification of the stance phases of the other three limbs, by coactivating flexors and extensors and/or by delaying push-off. Cutaneous reflexes during mid-stance were consistent with stabilizing the homonymous limb by delaying and then facilitating its push-off and modifying the support phases of the homolateral and diagonal limbs, characterized by coactivating flexors and extensors, reinforcing extensor activity and/or delaying push-off. The shortest latencies of homolateral and diagonal responses were consistent with fast-conducting disynaptic or trisynaptic pathways. Descending homolateral and diagonal pathways from the forelimbs to the hindlimbs had a higher probability of eliciting responses compared with ascending pathways from the hindlimbs to the forelimbs. Thus, in quadrupeds, intralimb and interlimb reflexes activated by cutaneous inputs ensure dynamic coordination of the four limbs, producing a whole-body response.SIGNIFICANCE STATEMENT The skin contains receptors that, when activated, send inputs to spinal circuits, signaling a perturbation. Rapid responses, or reflexes, in muscles of the contacted limb and opposite homologous limb help maintain balance and forward progression. Here, we investigated reflexes during quadrupedal locomotion in the cat by electrically stimulating cutaneous nerves in each of the four limbs. Functionally, responses appear to modify the trajectory or stabilize the movement of the stimulated limb while modifying the support phase of the other limbs. Reflexes between limbs are mediated by fast-conducting pathways that involve excitatory and inhibitory circuits controlling each limb. The comparatively stronger descending pathways from cervical to lumbar circuits controlling the forelimbs and hindlimbs, respectively, could serve a protective function.

Spinal control of muscle synergies for adult mammalian locomotion.

KEY POINTS: The control of locomotion is thought to be generated by activating groups of muscles that perform similar actions, which are termed muscle synergies. Here, we investigated if muscle synergies are controlled at the level of the spinal cord. We did this by comparing muscle activity in the legs of cats during stepping on a treadmill before and after a complete spinal transection that abolishes commands from the brain. We show that muscle synergies were maintained following spinal transection, validating the concept that muscle synergies for locomotion are primarily controlled by circuits of neurons within the spinal cord. ABSTRACT: Locomotion is thought to involve the sequential activation of functional modules or muscle synergies. Here, we tested the hypothesis that muscle synergies for locomotion are organized within the spinal cord. We recorded bursts of muscle activity in the same cats (n = 7) before and after spinal transection during tied-belt locomotion at three speeds and split-belt locomotion at three left-right speed differences. We identified seven muscles synergies before (intact state) and after (spinal state) spinal transection. The muscles comprising the different synergies were the same in the intact and spinal states as well as at different speeds or left-right speed differences. However, there were some significant shifts in the onsets and offsets of certain synergies as a function of state, speed and left-right speed differences. The most notable difference between the intact and spinal states was a change in the timing between the knee flexor and hip flexor muscle synergies. In the intact state, the knee flexor synergy preceded the hip flexor synergy, whereas in the spinal state both synergies occurred concurrently. Afferent inputs also appear important for the expression of some muscle synergies, specifically those involving biphasic patterns of muscle activity. We propose that muscle synergies for locomotion are primarily organized within the spinal cord, although their full expression and proper timing requires inputs from supraspinal structures and/or limb afferents.

Recovery of locomotion after spinal cord injury: some facts and mechanisms.

After spinal cord injury (SCI), various sensorimotor functions can recover, ranging from simple spinal reflexes to more elaborate motor patterns, such as locomotion. Locomotor recovery after complete spinalization (complete SCI) must depend on the presence of spinal circuitry capable of generating the complex sequential activation of various leg muscles. This is achieved by an intrinsic spinal circuitry, termed the central pattern generator (CPG), working in conjunction with sensory feedback from the legs. After SCI, different changes in cellular and circuit properties occur spontaneously and can be promoted by pharmacological, electrical, or rehabilitation strategies. After partial SCI, hindlimb locomotor recovery can result from regeneration or sprouting of spared pathways, but also from mechanisms observed after complete SCI, namely changes within the intrinsic spinal circuitry and sensory inputs.

Cutaneous sensory feedback from paw pads affects lateral balance control during split-belt locomotion in the cat.

Cutaneous sensory feedback from the paw pads plays an important role in regulating body balance, especially in challenging environments like ladder or slope walking. Here, we investigated the contribution of cutaneous sensory feedback from the paw pads to balance control in cats stepping on a split-belt treadmill. Forepaws and hindpaws were anesthetized unilaterally using lidocaine injections. We evaluated body balance in intact and compromised cutaneous feedback conditions during split-belt locomotion with belt-speed ratios of 0.5, 1.0, 1.5 and 2.0. Measures of body balance included step width, relative duration of limb support phases, lateral bias of center of mass (CoM) and margins of static and dynamic stability. In the intact condition, static and dynamic balance declined with increasing belt-speed ratio as a result of a lateral shift of the CoM toward the borders of support on the slower moving belt. Anesthesia of the ipsilateral paws improved locomotor balance with increasing belt-speed ratios by reversing the CoM shift, decreasing the relative duration of the two-limb support phase, increasing the duration of four- or three-limb support phases, and increasing the hindlimb step width and static stability. We observed no changes in most balance measures in anesthetized conditions during tied-belt locomotion at 0.4 m s-1 CoM lateral displacements closely resembled those of the inverted pendulum and of human walking. We propose that unilaterally compromised cutaneous feedback from the paw pads is compensated for by improving lateral balance and by shifting the body toward the anesthetized paws to increase tactile sensation during the stance phase.

Nonlinear Modulation of Cutaneous Reflexes with Increasing Speed of Locomotion in Spinal Cats.

Cutaneous reflexes are important for responding rapidly to perturbations, correcting limb trajectory, and strengthening support. During locomotion, they are modulated by phase to generate functionally appropriate responses. The goal of the present study was to determine whether cutaneous reflexes and their phase-dependent modulation are altered with increasing speed and if this is accomplished at the spinal level. Four adult cats that recovered stable hindlimb locomotion after spinal transection were implanted with electrodes to record hindlimb muscle activity chronically and to stimulate the superficial peroneal nerve electrically to evoke cutaneous reflexes. The speed-dependent modulation of cutaneous reflexes was assessed by evoking and characterizing ipsilateral and contralateral responses in semitendinosus, vastus lateralis, and lateral gastrocnemius muscles at four treadmill speeds: 0.2, 0.4, 0.6, and 0.8 m/s. The amplitudes of ipsilateral and contralateral responses were largest at intermediate speeds of 0.4 and 0.6 m/s, followed by the slowest and fastest speeds of 0.2 and 0.8 m/s, respectively. The phase-dependent modulation of reflexes was maintained across speeds, with ipsilateral and contralateral responses peaking during the stance-to-swing transition and swing phase of the ipsilateral limb or midstance of the contralateral limb. Reflex modulation across speeds also correlated with the spatial symmetry of the locomotor pattern, but not with temporal symmetry. That the cutaneous reflex amplitude in all muscles was similarly modulated with increasing speed independently of the background level of muscle activity is consistent with a generalized premotoneuronal spinal control mechanism that could help to stabilize the locomotor pattern when changing speed.SIGNIFICANCE STATEMENT When walking, receptors located in the skin respond to mechanical pressure and send signals to the CNS to correct the trajectory of the limb and to reinforce weight support. These signals produce different responses, or reflexes, if they occur when the foot is contacting the ground or in the air. This is known as phase-dependent modulation of reflexes. However, when walking at faster speeds, we do not know if and how these reflexes are changed. In the present study, we show that reflexes from the skin are modulated with speed and that this is controlled at the level of the spinal cord. This modulation could be important in preventing sensory signals from destabilizing the walking pattern.

The modulation of locomotor speed is maintained following partial denervation of ankle extensors in spinal cats.

Speed modulation requires spatiotemporal adjustments and altered neural drive to different muscles. The loss of certain muscles produces changes in the locomotor pattern and functional compensation. However, how the loss of specific muscles affects speed modulation has not been specifically investigated. Here, we denervated the lateral gastrocnemius and soleus muscles unilaterally in seven cats that had recovered hindlimb locomotion following complete spinal transection (spinal cats). Hindlimb locomotion was tested at 10 speeds, from 0.1 to 1.0 m/s, before, 1-2 days, and 1-8 wk after denervation. Six of seven cats performed hindlimb locomotion 1-2 days postdenervation at all speeds, with the exception of two out of those six cats that did not perform stable stepping at 0.9 and 1.0 m/s. All seven cats performed hindlimb locomotion 1-8 wk postdenervation at all speeds. In some cats, at 1-2 days postdenervation, the ipsilateral hindlimb performed more steps than the contralateral hindlimb, particularly at slow speeds. This 2:1 coordination disappeared over time. In three cats, the linear increase in the amplitude of the electromyography of the ipsilateral medial gastrocnemius was reduced with increasing speed early after denervation before recovering later on. Overall, the results indicate that spinal circuits interacting with sensory feedback from the hindlimbs compensate for the partial loss of ankle extensors, retaining the ability to modulate locomotor speed. NEW & NOTEWORTHY We investigated speed modulation after denervating 2 ankle extensors unilaterally at 10 treadmill speeds in spinal-transected cats. Although we observed new forms of left-right coordination and changes in muscle activity of a remaining synergist, modulation of spatiotemporal variables with increasing speed was largely maintained after denervation. The results indicate that spinal locomotor centers interacting with sensory feedback compensate for the loss of ankle extensors, allowing speed modulation.

Left-right coordination from simple to extreme conditions during split-belt locomotion in the chronic spinal adult cat.

KEY POINTS: Coordination between the left and right sides is essential for dynamic stability during locomotion. The immature or neonatal mammalian spinal cord can adjust to differences in speed between the left and right sides during split-belt locomotion by taking more steps on the fast side. We show that the adult mammalian spinal cord can also adjust its output so that the fast side can take more steps. During split-belt locomotion, only certain parts of the cycle are modified to adjust left-right coordination, primarily those associated with swing onset. When the fast limb takes more steps than the slow limb, strong left-right interactions persist. Therefore, the adult mammalian spinal cord has a remarkable adaptive capacity for left-right coordination, from simple to extreme conditions. ABSTRACT: Although left-right coordination is essential for locomotion, its control is poorly understood, particularly in adult mammals. To investigate the spinal control of left-right coordination, a spinal transection was performed in six adult cats that were then trained to recover hindlimb locomotion. Spinal cats performed tied-belt locomotion from 0.1 to 1.0 m s-1 and split-belt locomotion with low to high (1:1.25-10) slow/fast speed ratios. With the left hindlimb stepping at 0.1 m s-1 and the right hindlimb stepping from 0.2 to 1.0 m s-1 , 1:1, 1:2, 1:3, 1:4 and 1:5 left-right step relationships could appear. The appearance of 1:2+ relationships was not linearly dependent on the difference in speed between the slow and fast belts. The last step taken by the fast hindlimb displayed longer cycle, stance and swing durations and increased extensor activity, as the slow limb transitioned to swing. During split-belt locomotion with 1:1, 1:2 and 1:3 relationships, the timing of stance onset of the fast limb relative to the slow limb and placement of both limbs at contact were invariant with increasing slow/fast speed ratios. In contrast, the timing of stance onset of the slow limb relative to the fast limb and the placement of both limbs at swing onset were modulated with slow/fast speed ratios. Thus, left-right coordination is adjusted by modifying specific parts of the cycle. Results highlight the remarkable adaptive capacity of the adult mammalian spinal cord, providing insight into spinal mechanisms and sensory signals regulating left-right coordination.

Lack of adaptation during prolonged split-belt locomotion in the intact and spinal cat.

KEY POINTS: During split-belt locomotion in humans where one leg steps faster than the other, the symmetry of step lengths and double support periods of the slow and fast legs is gradually restored. When returning to tied-belt locomotion, there is an after-effect, with a reversal in the asymmetry observed in the early split-belt period, indicating that the new pattern was stored within the central nervous system. In this study, we investigated if intact and spinal-transected cats show a similar pattern of adaptation to split-belt locomotion by measuring kinematic variables and electromyography before, during and after 10 min of split-belt locomotion. The results show that cats do not adapt to prolonged split-belt locomotion. Our results suggest an important physiological difference in how cats and humans respond to prolonged asymmetric locomotion. ABSTRACT: In humans, gait adapts to prolonged walking on a split-belt treadmill, where one leg steps faster than the other, by gradually restoring the symmetry of interlimb kinematic variables, such as double support periods and step lengths, and by reducing muscle activity (EMG, electromyography). The adaptation is also characterized by reversing the asymmetry of interlimb variables observed during the early split-belt period when returning to tied-belt locomotion, termed an after-effect. To determine if cats adapt to prolonged split-belt locomotion and to assess if spinal locomotor circuits participate in the adaptation, we measured interlimb variables and EMG in intact and spinal-transected cats before, during and after 10 min of split-belt locomotion. In spinal cats, only the hindlimbs performed stepping with the forelimbs stationary. In intact and spinal cats, step lengths and double support periods were, on average, symmetric, during tied-belt locomotion. They became asymmetric during split-belt locomotion and remained asymmetric throughout the split-belt period. Upon returning to tied-belt locomotion, symmetry was immediately restored. In intact cats, the mean EMG amplitude of hindlimb extensors increased during split-belt locomotion and remained increased throughout the split-belt period, whereas in spinal cats, EMG amplitude did not change. Therefore, the results indicate that the locomotor pattern of cats does not adapt to prolonged split-belt locomotion, suggesting an important physiological difference in the control of locomotion between cats and humans. We propose that restoring left-right symmetry is not required to maintain balance during prolonged asymmetric locomotion in the cat, a quadruped, as opposed to human bipedal locomotion.

The neural control of interlimb coordination during mammalian locomotion.

Neuronal networks within the spinal cord directly control rhythmic movements of the arms/forelimbs and legs/hindlimbs during locomotion in mammals. For an effective locomotion, these networks must be flexibly coordinated to allow for various gait patterns and independent use of the arms/forelimbs. This coordination can be accomplished by mechanisms intrinsic to the spinal cord, somatosensory feedback from the limbs, and various supraspinal pathways. Incomplete spinal cord injury disrupts some of the pathways and structures involved in interlimb coordination, often leading to a disruption in the coordination between the arms/forelimbs and legs/hindlimbs in animal models and in humans. However, experimental spinal lesions in animal models to uncover the mechanisms coordinating the limbs have limitations due to compensatory mechanisms and strategies, redundant systems of control, and plasticity within remaining circuits. The purpose of this review is to provide a general overview and critical discussion of experimental studies that have investigated the neural mechanisms involved in coordinating the arms/forelimbs and legs/hindlimbs during mammalian locomotion.

Reflex wind-up in early chronic spinal injury: plasticity of motor outputs.

In this study we evaluate temporal summation (wind-up) of reflexes in select distal and proximal hindlimb muscles in response to repeated stimuli of the distal tibial or superficial peroneal nerves in cats 1 mo after complete spinal transection. This report is a continuation of our previous paper on reflex wind-up in the intact and acutely spinalized cat. To evaluate reflex wind-up in both studies, we recorded electromyographic signals from the following left hindlimb muscles: lateral gastrocnemius (LG), tibialis anterior (TA), semitendinosus (ST), and sartorius (Srt), in response to 10 electrical pulses to the tibial or superficial peroneal nerves. Two distinct components of the reflex responses were considered, a short-latency compound action potential (CAP) and a longer duration bout of sustained activity (SA). These two response types were shown to be differentially modified by acute spinal injury in our previous work (Frigon A, Johnson MD, Heckman CJ. J Physiol 590: 973-989, 2012). We show that these responses exhibit continued plasticity during the 1-mo recovery period following acute spinalization. During this early chronic phase, wind-up of SA responses returned to preinjury levels in one muscle, the ST, but remained depressed in all other muscles tested. In contrast, CAP response amplitudes, which were initially potentiated following acute transection, returned to preinjury levels in all muscles except for Srt, which continued to show marked increase. These findings illustrate that spinal elements exhibit considerable plasticity during the recovery process following spinal injury and highlight the importance of considering SA and CAP responses as distinct phenomena with unique underlying neural mechanisms.NEW & NOTEWORTHY This research is the first to assess temporal summation, also called wind-up, of muscle reflexes during the 1-mo recovery period following spinal injury. Our results show that two types of muscle reflex activity are differentially modulated 1 mo after spinal cord injury (SCI) and that spinal reflexes are altered in a muscle-specific manner during this critical period. This postinjury plasticity likely plays an important role in spasticity experienced by individuals with SCI.

Adaptive muscle plasticity of a remaining agonist following denervation of its close synergists in a model of complete spinal cord injury.

Complete spinal cord injury (SCI) alters the contractile properties of skeletal muscle, and although exercise can induce positive changes, it is unclear whether the remaining motor system can produce adaptive muscle plasticity in response to a subsequent peripheral nerve injury. To address this, the nerve supplying the lateral gastrocnemius (LG) and soleus muscles was sectioned unilaterally in four cats that had recovered hindlimb locomotion after spinal transection. In these spinal cats, kinematics and electromyography (EMG) were collected before and for 8 wk after denervation. Muscle histology was performed on LG and medial gastrocnemius (MG) bilaterally in four spinal and four intact cats. In spinal cats, cycle duration for the hindlimb ipsilateral or contralateral to the denervation could be significantly increased or decreased compared with predenervation values. Stance duration was generally increased and decreased for the contralateral and ipsilateral hindlimbs, respectively. The EMG amplitude of MG was significantly increased bilaterally after denervation and remained elevated 8 wk after denervation. In spinal cats the ipsilateral LG was significantly smaller than the contralateral LG, whereas the ipsilateral MG weighed significantly more than the contralateral MG. Histological characterizations revealed significantly larger fiber areas for type IIa fibers of the ipsilateral MG in three of four spinal cats. Microvascular density in the ipsilateral MG was significantly higher than in the contralateral MG. In intact cats, no differences were found for muscle weight, fiber area, or microvascular density between homologous muscles. Therefore, the remaining motor system after complete SCI retains the ability to produce adaptive muscle plasticity.