Force regulation of ankle extensor muscle activity in freely walking cats.

To gain insight into the relative importance of force feedback to ongoing ankle extensor activity during walking in the conscious cat, we isolated the medial gastrocnemius muscle (MG) by denervating the other ankle extensors and measured the magnitude of its activity at different muscle lengths, velocities, and forces accomplished by having the animals walk up and down a sloped pegway. Mathematical models of proprioceptor dynamics predicted afferent activity and revealed that the changes in muscle activity under our experimental conditions were strongly correlated with Ib activity and not consistently associated with changes in Ia or group II activity. This allowed us to determine the gains within the force feedback pathway using a simple model of the neuromuscular system and the measured relationship between MG activity and force. Loop gain increased with muscle length due to the intrinsic force-length property of muscle. The gain of the pathway that converts muscle force to motoneuron depolarization was independent of length. To better test for a causal relationship between modulation of force feedback and changes in muscle activity, a second set of experiments was performed in which the MG muscle was perturbed during ground contact of the hind foot by dropping or lifting the peg underfoot. Collectively, these investigations support a causal role for force feedback and indicate that about 30% of the total muscle activity is due to force feedback during level walking. Force feedback's role increases during upslope walking and decreases during downslope walking, providing a simple mechanism for compensating for changes in terrain.

Role of sensory feedback in the control of stance duration in walking cats.

The rate of stepping in the hind legs of chronic spinal and decerebrate cats adapts to the speed of the treadmill on which the animals walk. This adaptive behavior depends on sensory signals generated near the end of stance phase controlling the transition from stance to swing. Two sensory signals have been identified to have this role: one from afferents activated by hip extension, most likely arising from muscle spindles in hip flexor muscles, and the other from group Ib afferents from Golgi tendon organs in the ankle extensor muscles. The relative importance of these two signals in controlling the stance to swing transition differs in chronic spinal cats and in decerebrate cats. Activation of hip afferents is necessary for controlling the transition in chronic spinal cats but not in decerebrate cats, while reduction in activity in group Ib afferents from GTOs is the primary factor controlling the transition in decerebrate cats. Possible mechanisms for this difference are discussed. The extent to which these two sensory signals control the stance to swing transition in normal walking cats is unknown, but it is likely that both could play an important role when animals are walking in a variable environment.

Contextual learning and obstacle memory in the walking cat.

Animals in their natural environments display a remarkably diverse variety of walking patterns. Although some of this diversity is generated by alterations in feedback from the moving limbs, animals can modify their walking in many ways that cannot be directly attributed to this sensory feedback. For example, animals and humans can learn to associate a particular environment with disturbances that were experienced there earlier, and alter their stepping accordingly even after the disturbance has ceased. Another relevant example is that walking animals are aware of the locations of obstacles around them, and use this awareness to alter their stepping patterns even when there is no visual information available about the location of the obstacles relative to the body. In this article, we discuss recent work from our laboratory that addresses these two topics. First, we report that perturbing walking cats in a consistent manner evokes long-lasting changes to the walking pattern that are expressed only in the context in which walking was disturbed. Secondly, we show that cats that have stepped over an obstacle remember the location of that obstacle relative to the body during long delays, and can use that memory to guide stepping. The general theme of this research is that sensory inputs that signal context-the visual and auditory environment that surrounds an animal-play an important role in shaping the basic pattern of locomotion.

Long-lasting, context-dependent modification of stepping in the cat after repeated stumbling-corrective responses.

A consistent feature of animal locomotion is the capacity to maintain stable movements in changing environments. Here we describe long-term modification of the swing movement of the hind leg in cats in response to repeatedly impeding the movement of the leg. While studying phase transitions in the hind legs, we discovered that repetitively evoking the stumbling-corrective reaction led to long-lasting increases in knee flexion and step height during swing to avoid the impediment. These increases were apparent after nearly 20 stimuli and maximal after about 120 stimuli and, in some animals, they persisted for > or =24 h after presentation of the stimuli. Furthermore, these long-lasting changes were context dependent and did not generalize to other environments; when walking was observed in an environment distinct from that used in training, the hind-limb kinematics returned to normal. To gain insight into what regions of the nervous system might be involved in this long-term modification, we examined the changes in stepping in decerebrate cats after multiple perturbed steps. In this situation, there was a short-term increase in step height, although this increase was smaller than that evoked in intact animals and persisted for <1 min after termination of the stimuli. Thus induction of the long-term increase in step height requires the forebrain. We propose that the conditioned change in leg movement is related to a general ability of animals to adapt locomotor movements to new features of the environment.

Coordination of fore and hind leg stepping in cats on a transversely-split treadmill.

To gain insight into the mechanism of coordination of stepping in the fore and hind legs of quadrupeds, we examined the kinematics of leg movements and the motor patterns in fore and hind leg flexor muscles in decerebrate walking cats when the two pairs of legs stepped on separate treadmills running at different speeds. When the front treadmill was slowed progressively from 0.6 to 0.3 m/s with the rear treadmill running at 0.6 m/s, the rate of stepping in both the fore and hind legs decreased and a 1:1 stepping ratio was maintained. The decrease in the rate of stepping in the hind legs was due primarily to an increase in the duration of the swing phase. Slowing the speed of the rear treadmill while keeping the front treadmill speed at 0.6 m/s decreased the rate of stepping of the hind legs, but had relatively little influence on the average rate of stepping in the forelegs. In this situation stepping in the fore and hind legs was uncoupled and the time of stepping in one hind leg relative to the ipsilateral foreleg progressively shifted during a walking sequence. Analysis of the timing of electromyographic (EMG) recordings from flexor muscles of the hip and elbow joints yielded insight into the neuronal mechanisms underlying the asymmetry in slowing either the front or rear treadmill. We propose that ipsilateral pattern generating networks are asymmetrically coupled via descending inhibitory pathways and an ascending excitatory pathway. We discuss how the characteristics of these linkages are functionally appropriate for establishing the normal timing of stepping in the hind and forelegs during slow walking.

Long-lasting memories of obstacles guide leg movements in the walking cat.

We examined the ways in which memories of previously seen obstacles can alter the stepping of walking cats. Cats were paused after the forelegs, but not the hindlegs, had stepped over an obstacle. Near the beginning of a variable delay period, the obstacle was lowered. On the subsequent step, the path of the hindlegs allowed us to make inferences about whether the memory of the obstacle was influencing leg movements. We present two main findings. First, the memory of the obstacle persisted for the duration that the animal straddled the original location of the obstacle. In one instance, this interval was 10 min. Second, this memory includes information regarding the size and position of the obstacle relative to the animal. This information is used to plan foot placement and to redirect the step in mid-swing to avoid the previous position of the obstacle.

Recruitment of gastrocnemius muscles during the swing phase of stepping following partial denervation of knee flexor muscles in the cat.

In walking cats, the biarticular medial and lateral gastrocnemius (MG-LG) muscles act to produce extension and flexion torques at the ankle and knee, respectively, and they usually display only one burst of activity beginning just before ground contact and ending near the end of the stance phase. Currently, the MG-LG muscles are considered to function primarily to control extension movements around the ankle joint during the stance phase. However, their flexion action at the knee means that they have the capacity to regulate rotations at the knee, but this role has not yet been clearly defined. Following partial denervation of the other muscles that normally act to flex the knee during swing, we observed that the MG-LG muscles, but not the Soleus muscle (a pure ankle extensor), often generated strong bursts of activity during early swing. These bursts were enhanced following mechanical stimulation of the paw, and they were especially prominent when the leg trailed over an object. They were absent when the leg led over an object. During treadmill walking the swing-related bursts in MG and LG had little influence on ankle flexion at the beginning of swing, but they were associated with slowing of ankle flexion when the leg trailed over an object. We hypothesized that the recruitment of these bursts functions to partially compensate for the reduction in knee torque resulting from the denervation of other knee flexors. Consistent with this hypothesis was our finding that the magnitude of the swing-related activity in the MG-LG muscles was linearly correlated to the extent of the knee flexion and to the peak angular velocity of knee flexion, and that the timing of the bursts was similar to that in the denervated muscles prior to denervation. Our findings suggest that an excitatory pathway exists from the flexor half-center of the central pattern-generating network to MG-LG motoneurons, and that this pathway is strongly regulated by central and/or peripheral signals.

A role for hip position in initiating the swing-to-stance transition in walking cats.

In this investigation, we obtained data that support the hypothesis that afferent signals associated with hip flexion play a role in initiating the swing-to-stance transition of the hind legs in walking cats. Direct evidence came from observations in walking decerebrate cats. Assisting the flexion of the hip joint during swing advanced the onset of activity in ankle extensor muscles, and this advance was strongly correlated with a reduction in the duration of hip flexor muscle activity. The hip angle at the time of onset of the flexion to extension transition was similar during assisted and unassisted steps. Additional evidence for the hypothesis that sensory signals related to hip flexion are important in regulating the swing-to-stance transition came from four normal animals trained to walk in a variety of situations designed to alter the coordination of movements at the hip, knee, and ankle joints during the swing phase. Although there were exceptions in some tasks and preparations, the angle of the hip joint at the time of onset of extensor activity was generally less variable than that of the knee and ankle joints. We also found no clear relationships between the angle of the limb and body axes, or the length of the limb axis, and the time of onset of extensor activity. Finally, there were no indications that the stretching of ankle extensor muscles during swing was a factor in regulating the transition from swing-to-stance.

A new electrode configuration for recording electromyographic activity in behaving mice.

With the increasing use of normal and genetically modified mice in the field of motor physiology, there is a need for a simple and reliable technique for recording electromyographic (EMG) activity in behaving mice. Here, we describe a new method for the fabrication and implantation of fine EMG recording electrodes into multiple muscles of adult mice. This method minimizes surgical damage to the muscles and the connecting leads have only a modest influence on leg movements when electrodes are implanted into distal muscles. We demonstrate that excellent EMG recordings can be obtained during walking, swimming and scratching for the vastus lateralis, tibialis anterior and gastrocnemius muscles in normal adult mice. EMG recordings were also made in a mutant EphA4 mouse to demonstrate the utility of the method for examining motor patterns in genetically modified animals. We also developed a method for constructing highly reflective markers that could be viewed over a range of orientations to measure the kinematics of legs movements during stepping. A commercial motion analysis system was used to track six markers during walking and to synchronize video and EMG data during walking sequences.

Contribution of force feedback to ankle extensor activity in decerebrate walking cats.

Previous investigations have demonstrated that feedback from ankle extensor group Ib afferents, arising from force-sensitive Golgi tendon organs, contributes to ankle extensor activity during the stance phase of walking in the cat. The objective of this investigation was to gain insight into the magnitude of this contribution by determining the loop gain of the positive force feedback pathway. Loop gain is the relative contribution of force feedback to total muscle activity and force. In decerebrate cats, the isolated medial gastrocnemius muscle (MG) was held at different lengths during sequences of rhythmic contractions associated with walking in the other three legs. We found that MG muscle activity and force increased at longer muscle lengths. A number of observations indicated that this length dependence was not due to feedback from muscle spindles. In particular, activity in group Ia afferents was insensitive to changes in muscle length during the MG bursts, and electrical stimulation of group II afferents had no influence on the magnitude of burst activity in other ankle extensors. We concluded that the homonymous positive force feedback pathway was isolated from other afferent pathways, allowing the use of a simple model of the neuromuscular system to estimate the pathway loop gain. This gain ranged from 0.2 at short muscle lengths to 0.5 at longer muscle lengths, demonstrating that force feedback was of modest importance at short muscle lengths, accounting for 20% of total activity and force, and of substantial importance at long muscle lengths, accounting for 50%. This length dependence was due to the intrinsic force-length property of muscle. The gain of the pathway that converts muscle force to motoneuron depolarization was independent of length. We discuss the relevance of this conclusion to the generation of ankle extensor activity in intact walking cats. These findings emphasize the general importance of feedback in generating ankle extensor activity during walking in the cat.

Chemical ablation of sensory afferents in the walking system of the cat abolishes the capacity for functional recovery after peripheral nerve lesions.

Weakening the ankle extensor muscles of cats by denervation of the synergists of the medial gastrocnemius (MG) muscle results in transient increase in yield at the ankle during early stance. Recovery of ankle function occurs over a period of 1-2 weeks, is use-dependent, and is associated with increases in the strength of reflexes from MG group I muscle afferents and an increase in the magnitude of bursts in the MG muscles during stance. These observations have led to the hypothesis that feedback from large muscle afferents is necessary for functional recovery. In this investigation we have tested this hypothesis by examining functional recovery in animals treated with pyridoxine, a drug known to destroy large muscle afferents. In four adult animals we confirmed that pyridoxine abolished the group I-mediated tendon-tap reflex in the ankle extensor muscle, and subsequently found that group I afferents from MG were either destroyed or non-conducting. Immediately after pyridoxine treatment the animals showed severe locomotor dysfunction but all recovered significantly over a period of 1 or 2 months and showed only minor kinematics deficits at the time of the muscle denervations. In all four pyridoxine-treated animals, weakening of the ankle extensors by denervation of the synergists of the MG muscle resulted in a large increase in yield at the ankle that persisted almost unchanged for a month after the operation. The magnitude of burst activity in the MG muscle during early stance of the pyridoxine-treated animals either did not increase or increased only slightly after the denervation of synergists. These observations are consistent with the hypothesis that feedback from group I afferents is necessary for functional recovery in untreated animals.

Toe flexor muscle spindle discharge and stretch modulation during locomotor activity in the decerebrate cat.

In order to investigate the nature (i.e. static or dynamic) of fusimotor drive to the flexor hallucis longus (FHL) and flexor digitorum longus (FDL) muscles during locomotion we recorded Ia and group II muscle spindle afferent responses to sinusoidal stretch (0.25 and 1 mm amplitude, respectively, 4-5 Hz) in a decerebrate cat preparation. FHL Ia and group II afferents generally had increased discharge rates and decreased modulation to stretch throughout the step cycle, compared to rest, suggesting raised static gamma drive at all locomotor phases. Although the modulation of Ia afferents was reduced during locomotion, most (13 of 18) showed a clear increasing trend during homonymous muscle activity (extension). This was consistent with phasic dynamic gamma drive to FHL spindles linked with alpha drive. In agreement with previous reports, FHL gave a single burst of EMG activity during the step cycle while FDL alpha drive had two components. One was related to extension while the other comprised a brief burst around the end of this phase. Typically FDL Ia and group II afferents also had elevated firing rates and reduced modulation at all locomotor phases, again implicating static gamma drive. Half the afferents (seven Ia, three group II) showed increased discharge during extension, suggesting phasic static gamma drive. There was no gamma drive associated with the late FDL alpha burst. In conclusion, the gamma drives to FHL and FDL differed during locomotion. FHL, which has the alpha drive of a classic extensor, received gamma drive that closely resembled other extensors. The gamma drive of FDL, which exhibits both extensor and flexor alpha synergies, did not match either muscle type. These observations are compatible with the view that fusimotor drive varies in different muscles during locomotion according to the prevailing sensorimotor requirements.

Adaptive changes in locomotor activity following botulinum toxin injection in ankle extensor muscles of cats.

The present study investigated the adaptations made in motor behavior following a temporary reduction in ankle extensor activity in the walking cat. Temporary muscle weakness was induced by injecting botulinum toxin into the lateral gastrocnemius (LG), plantaris (PL), and soleus (SOL) muscles, or SOL alone. The medial gastrocnemius (MG) muscle was not injected. Adaptations in the level of muscle activity were recorded using chronically implanted electromyographic (EMG) electrodes. Serial recordings were made prior to botulinum toxin injections and for several days following the injections. Kinematic analysis of ankle joint movements was made from video records to assess the impact of the botulinum toxin injections on the function of the ankle joint during walking. Following injection of the LG, PL, and SOL muscles with botulinum toxin, the amplitude of the MG burst increased over a period of a few days to a week. This increase was similar to the previously reported changes produced in MG following transection of the nerves serving LG, PL, and SOL. Following the weakening of the ankle extensor muscles, there was a temporary deficit in ankle function during walking as evidenced by a marked increase in the amount of ankle flexion that occurred at stance onset. This functional deficit recovered relatively quickly and was not associated with a return of the EMG pattern to the preinjection pattern. After recovery from the initial injections, a second injection of botulinum toxin into SOL alone was performed. No functional deficits were observed in the ankle movements during walking following this second injection. However, weakening SOL produced increases in the burst amplitudes of the MG, LG, and PL muscles over a period of a few days. This suggests that normal movements at the ankle during walking can be generated with more than one pattern of ankle extensor activity and that there is flexibility in how the necessary torque is produced. A final procedure, transection of the nerves serving LG, PL, and SOL, failed to produce any functional deficits in ankle movements. The implication is that adaptations to the neural control of ankle extensor activity that were induced by the initial procedure persisted after the recovery of the injected muscles and were sufficient to compensate for the subsequent challenges.

Proprioceptive modulation of hip flexor activity during the swing phase of locomotion in decerebrate cats.

This study examined the influence of proprioceptive input from hip flexor muscles on the activity in hip flexors during the swing phase of walking in the decerebrate cat. One hindlimb was partially denervated to remove cutaneous input and afferent input from most other hindlimb muscles. Perturbations to hip movement were applied either by 1) manual resistance or assistance to swing or by 2) resistance to hip flexion using a device that blocked hip flexion but allowed leg extension. Electromyographic recordings were made from the iliopsoas (IP), sartorius, and medial gastrocnemius muscles. When the hip was manually assisted into flexion, there was a reduction in hip flexor burst activity. Conversely, when hip flexion was manually resisted or mechanically blocked during swing, the duration and amplitude of hip flexor activity was increased. We also found some specificity in the role of afferents from individual hip flexor muscles in the modulation of flexor burst activity. If the IP muscle was detached from its insertion, little change in the response to blocking flexion was observed. Specific activation of IP afferent fibers by stretching the muscle also did not greatly affect flexor activity. On the other hand, if conduction in the sartorius nerves was blocked, there was a diminished response to blocking hip flexion. The increase in duration of the flexor bursts still occurred, but this increase was consistently lower than that observed when the sartorius nerves were intact. From these results we propose that during swing, feedback from hip flexor muscle afferents, particularly those from the sartorius muscles, enhances flexor activity. In addition, if we delayed the onset of flexor activity in the contralateral hindlimb, blocking hip flexion often resulted in the prolongation of ipsilateral flexor activity for long periods of time, further revealing the reinforcing effects of flexor afferent feedback on flexor activity. This effect was not seen if conduction in the sartorius nerves was blocked. In conclusion, we have found that hip flexor activity during locomotion can be strongly modulated by modifying proprioceptive feedback from the hip flexor muscles.

Could enhanced reflex function contribute to improving locomotion after spinal cord repair?

Although numerous treatments have been found to improve locomotion in spinal cord injured mammals, the underlying mechanisms are very poorly understood. Some of the main possibilities are: (1) regeneration of axons across the injury site and the re-establishment of descending pathways needed to voluntarily initiate and maintain stepping in the hind legs, (2) enhanced effectiveness of undamaged neurons in preparations with incomplete transections of the cord, (3) non-specific facilitation of reflexes and intrinsic spinal networks by transmitters released from regenerated axons and/or by substances introduced by the treatment, and (4) enhanced trunk movements close to the injury site strengthening the mechanical coupling of the trunk to the hind legs via spinal reflexes. In addition, any procedure that even slightly improves stepping may be further enhanced by use-dependent modification of reflex pathways and interneuronal networks in the lumbar cord. The emphasis of this review is on the contribution of spinal reflexes to the patterning of motor activity for walking, and how enhancing reflex function may contribute to the improvement of locomotion by treatments aimed at restoring locomotion after complete transection of the spinal cord.

Adaptive locomotor plasticity in chronic spinal cats after ankle extensors neurectomy.

After lateral gastrocnemius-soleus (LGS) nerve section in intact cats, a rapid locomotor compensation involving synergistic muscles occurs and is accompanied by spinal reflex changes. Only some of these changes are maintained after acute spinalization, indicating the involvement of descending pathways in functional recovery. Here, we address whether the development of these adaptive changes is dependent on descending pathways. The left LGS nerve was cut in three chronic spinal cats. Combined kinematics and electromyographic (EMG) recordings were obtained before and for 8 d after the neurectomy. An increased yield at the ankle was present early after neurectomy and, as in nonspinal cats, was gradually reduced within 8 d. Compensation involved transient changes in step cycle structure and a longer term increase in postcontact medial gastrocnemius (MG) EMG activity. Precontact MG EMG only increased in one of three cats. In a terminal experiment, the influence of group I afferents from MG and LGS on stance duration was measured in two cats. LGS effectiveness at increasing stance duration was largely decreased in both cats. MG effectiveness was only slightly changed: increased in one cat and decreased in another. In cat 3, the plantaris nerve was cut after LGS recovery. The recovery time courses from both neurectomies were similar (p > 0.8), suggesting that this spinal compensation is likely a generalizable adaptive strategy. From a functional perspective, the spinal cord therefore must be considered capable of adaptive locomotor plasticity after motor nerve lesions. This finding is of prime importance to the understanding of functional plasticity after spinal injury.

Use-dependent gain change in the reflex contribution to extensor activity in walking cats.

Denervation of the synergists of the medial gastrocnemius (MG) muscle in the cat hind leg results in a progressive increase in the magnitude of burst activity in the MG muscle during walking. The increase in burst magnitude is associated with an increase in the slope of the relationship between the magnitude of individual MG bursts and the amplitude of ankle flexion during stance. This finding is consistent with the hypothesis that the increase in MG burst magnitude is due to an increase in gain of reflex pathways reinforcing the activation of MG. The increase in slope is use-dependent since it was not observed when the leg was released from a cast that immobilized the leg for 6 days.

Functional role of muscle reflexes for force generation in the decerebrate walking cat.

To quantify the importance of reflexes due to muscle length changes in generating force during walking, we studied high decerebrate cats that walked on a treadmill. One leg was denervated except for the triceps surae and a few other selected muscles. The triceps surae muscles are ankle extensor muscles that attach to the Achilles' tendon which was cut and connected to a muscle puller. In some steps the EMG activity triggered the puller to move the muscle through the pattern of length changes that are normally produced by ankle movements in intact cats walking over ground (simulated walking). In other steps the muscles were held isometrically. The EMG and force produced during the two types of steps were compared. On average about 50 % more EMG was generated during the E2 part of the simulated stance phase in the triceps surae muscles, but not in other muscles studied. Force was increased significantly over the entire stance phase by about 20 %, when muscle stretches simulating walking were applied. However, during much of the stance phase the triceps surae muscles are shortening and so would produce less force. The effect of shortening was assessed in control experiments in which these muscles were stimulated at a constant frequency, either isometrically or during simulated walking movements. By combining data from the walking and control experiments, we estimate that about 35 % of the force produced in the cat ankle extensors during stance is produced by reflexes due to muscle length changes. Other sensory inputs may also contribute to force production, but the total reflex contribution will vary under different conditions of speed, length, loading, task difficulty, etc. Since a substantial percentage of the force in the stance phase of walking is normally produced by muscle reflexes, this force can be continuously adjusted up or down, if the muscles receive extra stretch or unloading during a particular step cycle.