Locomotor Stepping Research Articles

Spinal Epidural Stimulation Strategies: Clinical Implications of Locomotor Studies in Spinal Rats

Significant advancements in spinal epidural stimulation (ES) strategies to enable volitional motor control in persons with a complete spinal cord injury (SCI) have generated much excitement in the field of neurorehabilitation. Still, an obvious gap lies in the ability of ES to effectively generate a robust locomotor stepping response after a complete SCI in rodents, but not in humans. In order to reveal potential discrepancies between rodent and human studies that account for this void, in this review, we summarize the findings of studies that have utilized ES strategies to enable successful hindlimb stepping in spinal rats. Recent clinical and preclinical evidence indicates that motor training with ES plays a crucial role in tuning spinal neural circuitry to generate meaningful motor output. Concurrently administered pharmacology can also facilitate the circuitry to provide near optimal motor performance in SCI rats. However, as of today, the evidence for pharmacological agents to enhance motor function in persons with complete SCI is insignificant. These and other recent findings discussed in this review provide insight into addressing the translational gap, guide the design of relevant preclinical experiments, and facilitate development of new approaches for motor recovery in patients with complete SCIs.

Effects of Induced Astigmatism on Foot Placement Strategies when Stepping onto a Raised Surface

PurposeLarge changes in spectacle prescription can increase falls risk in older people. We investigated the effect of induced astigmatism (a common cause of distorted or blurred vision in older people) on locomotor stepping patterns to determine whether the orientation of astigmatic changes could have differential effects on gait safety when negotiating steps and stairs.Methods10 older adults (mean age 76.0±6.4 years) walked up to and stepped onto a raised block whilst wearing their spectacle prescription and when blurred with ±3.00D cylinders at axes 45°, 90°, 135° and 180°. Gait measurements included foot placement before the block, toe clearance over the block edge and foot placement on the block.ResultsInduced astigmatism with axes at 90°, providing magnification in the horizontal meridian only, caused no change in stepping pattern. Induced astigmatism with axes at 180° caused foot placement changes in the anterior or posterior direction according to whether magnification was positive or negative in the vertical meridian (block perceived higher or lower respectively). Induced astigmatism with axes oblique at 45° and 135° (causing the block to be perceived as a parallelogram sloping downwards either to the right or left) caused gait changes in the anterior and posterior, vertical and lateral directions. Changes in lateral foot placement appeared to be an attempt to maintain constant foot clearance levels over the block edge by stepping over the perceived ‘lower’ side of the ‘sloping’ block.ConclusionsAstigmatic changes with oblique axes had the greatest effect on gait. Clinicians, including optometrists, physiotherapists, occupational therapists and nurses should counsel older patients about the effects of astigmatism on gait safety. Furthermore, partial prescribing of astigmatic corrections should be considered to reduce the risk of falling.

Protective stepping response in Parkinsonian patients and the effect of vibrotactile feedback

The objectives of this study were (1) to characterize protective stepping responses to unpredictable forward/backward postural perturbation in Parkinson's disease (PD) and (2) to assess whether vibrotactile cues of the impending fall improve the stepping response. Twenty mild PD patients, 7 advanced PD patients, and 17 age-matched controls stood on a platform moving unpredictably forward and backward, requiring a protective step to maintain balance. Direction-coded vibrotactile cues, triggered by leg tilt, were provided to prompt step generation. All subjects showed quicker reaction time, shorter steps, and smaller total trunk displacement when stepping backward than when stepping forward. Advanced PD patients took shorter, slower, and an increased number of protective steps. The only abnormality observed in mild PD patients was slightly slower backward steps. Vibrotactile feedback reduced the amount of trunk displacement observed before taking a protective step but did not improve any abnormality in PD patients. Early PD patients had near-normal protective stepping responses to unpredictable perturbations, but advanced patients made slow and short steps both forward and backward. Given that latencies were preserved even in unpredictable conditions, step slowness and hypometria are the primary abnormalities of the stepping response in PD. As voluntary locomotor stepping in PD is reported to improve with sensory feedback, the lack of such improvement in our study implies that additional sensory cues cannot help automatic reflex-like stepping reactions in PD patients.

X-ray kinematics analysis of vaginal scent marking in female Syrian hamsters (Mesocricetus auratus)

Vaginal marking is a stereotyped scent marking behavior in female Syrian hamsters used to attract male hamsters for mating. Although the modulation of vaginal marking by hormones and odors is well understood, the motor control of this proceptive reproductive behavior remains unknown. Therefore, we used X-ray videography to visualize individual bone movements during vaginal marking. Kinematic analyses revealed several consistent motor patterns of vaginal marking. Despite exhibiting a diversity of trial-to-trial non-marking behaviors (e.g. locomotor stepping), we found that lowering and raising the pelvis consistently corresponded with coordinated flexion and extension cycles of the hip, knee, and tail, suggesting that these movements are fundamental to vaginal marking behavior. Surprisingly, we observed only small changes in the angles of the pelvic and sacral regions, suggesting previous reports of pelvic rotation during vaginal marking may need to be reconsidered. From these kinematic data, we inferred that vaginal marking is primarily due to the actions of hip and knee extensor muscles of the trailing leg working against gravity to support the weight of the animal as it controls the descent of the pelvis to the ground. The cutaneous trunci muscle likely mediates the characteristic flexion of the tail. Interestingly, this tail movement occurred on the same time scale as the joint kinematics suggesting possible synergistic recruitment of these muscle groups. These data therefore provide new targets for future studies examining the peripheral control of female reproductive behaviors.

Brainstem regions with neuronal activity patterns correlated with priming of locomotor stepping in the anesthetized rat

Locomotor stimulation in the perifornical hypothalamus produces a transient facilitation of subsequent locomotion, a priming effect, such that stepping to a second train of stimulation occurs with a shorter latency of onset and increased amplitude. Neurons responsible for the initiation of this facilitated stepping presumably respond to locomotor stimulation with a similar priming effect, i.e. either a shorter latency or a larger change in activity rate. This study used anesthetized rats (urethane, 800 mg/kg) to compare brainstem regions in terms of the relative rates of occurrence of single neurons that showed both specific responses to locomotor stimulation and also priming effects. Specific responses were characterized by a progressive increase in activity prior to the first step (a Type I pattern). In that they co-varied in time with the increased probability of stepping onset, Type I responses were more specific than Type II responses, which peaked early in the stimulation train several seconds before the onset of stepping. Regions with high proportions of neurons showing Type I responses and priming effects included the anterior dorsal tegmentum lateral to the central gray, the oral pontine reticular nucleus and the medial gigantocellular nucleus. Few Type I neurons showed a modulation of activity related to the step cycle. Type I primed neurons were uncommon in the cuneiform and the pedunculopontine regions, but neurons showing other patterns (decreases and antidromic responses) were relatively prevalent there. The ventral tegmental area was generally unresponsive. The results indicate that stepping elicited by perifornical stimulation in the anesthetized rat is mediated by circuits that differ at midbrain levels from the circuits implicated in other types of locomotion. Two regions, the anterior dorsal tegmentum and the oral pontine reticular nucleus, warrant further attention to determine their possible roles in the initiation of locomotion.

Priming pattern determines the correlation between hippocampal theta activity and locomotor stepping elicited by stimulation in anesthetized rats

The after-effects of locomotor stimulation are a transient facilitation of locomotor initiation (the priming effect), and a transient increase in hippocampal rhythmic slow activity in the 3–6 Hz band of the theta range. The similar time course of the two effects suggests that hippocampal 3–6 Hz activity may be linked to the excitability of locomotor initiation. This study tested the hypothesis that power in the 3–6 Hz band that is present prior to stimulation would predict the magnitude of elicited stepping. Stimulation electrodes were implanted in 15 locomotor sites of 10 anesthetized rats (urethane, 800 mg/kg). Hindlimb stepping was elicited by a single control train of electrical stimulation presented once every 62 s. On test trials, a test train at the same intensity followed the control train at varying control/test intervals (15–36 s) to assess the priming effect on stepping. The priming pattern determined whether hippocampal 3–6 Hz power predicted the amount of stepping to be elicited by a stimulation train. Positive correlation (0.47> r>0.22) was found for seven out of eight sites showing positive priming effects. Correlation was absent for three other sites that showed non-significant priming effects and were mixed for four sites that showed negative effects. Sites with positive priming patterns, compared to sites with inconsistent or negative priming patterns, had similar trends in post-stimulation 3–6 Hz power, smaller increases in 6–8 Hz power during the control train and lower 1–3 Hz power during the periods immediately before the control stimulation. For six of 15 sites, regardless of the priming pattern, 1–3 Hz power was inversely related to subsequent stepping, and in three cases provided an independent predictor of stepping. Stimulation at two sites produced discrete episodes of post-stimulation stepping. In one of these cases, a 0.5-Hz increase in peak frequency of hippocampal activity preceded stepping. The results show that the association between hippocampal 3–6 Hz activity and the excitability of locomotor initiation is sufficiently specific to allow prediction of the magnitude of stepping by the prior levels of 3–6 Hz power. However, the occurrence of negative priming effects during prominent 3–6 Hz activity indicates that other factors determine the actual stepping and they can suppress the correlation between theta activity and subsequent locomotion.

Hippocampal theta activity and facilitated locomotor stepping produced by GABA injections in the midbrain raphe region

Inactivation of neurons in the midbrain raphe region produces increases in locomotor activity, and it appears that they function to suppress locomotion. Inactivation of neurons there also produces hippocampal slow wave (theta) activity and it appears that they also function to inhibit rhythmic activity in the hippocampus. We determined whether the degree of association between the two effects was consistent with the operation of a single mechanism. Stimulation electrodes were implanted into locomotor sites of the hypothalamus of 34 urethane-anesthetized rats. Hindlimb stepping was elicited by 5.12-s trains of perifornical electrical stimulation presented once per minute. Hippocampal theta activity was recorded across the CA1 layer of the dorsal hippocampus. GABA injections were used to locate raphe sites at which neuronal inactivation influenced stepping and hippocampal activity. A glass pipette (80-μm tip) was inserted into the midbrain, and injections of GABA (50–100 mg/0.1–0.2 μl) were made in 70 sites in the midbrain. Injections at 34 sites facilitated stimulation-elicited stepping, and at 17 sites, they also produced intertrial stepping. Facilitating injections, but not ineffective or suppressive injections, increased the mean peak frequency of hippocampal activity, and increased power in the 4–5 Hz band during the period that preceded the stimulation trains, but did not change the 5–6 Hz activity produced during the stimulation trains. Priming locomotor stimulation which also facilitated stepping produced generally similar increases in pre-stimulation peak frequency and 4-Hz power. The magnitudes of the increases in stepping and 4-Hz power were uncorrelated. The increase in 4-Hz power appeared earlier than the increase in stepping in 18 of 34 cases, and later in 11 cases; no increases in 4-Hz power were apparent in five cases. The results indicate that pre-locomotor 4-Hz hippocampal activity in the urethane-anesthetized rat is loosely coupled with facilitated locomotor initiation. Neurons in the midbrain raphe region appear to suppress both processes, but the low degree of association between the magnitudes and onset times of increases in stepping and hippocampal 4-Hz power indicate the operation of multiple mechanisms.

Locomotion and head scanning initiated by hypothalamic stimulation are inversely related

Stimulation in the hypothalamus elicits locomotor stepping. Before stepping is initiated, head scanning movements occur. We determined the relationships between the latency of locomotor initiation and the number, extent and direction of the head scanning movements. Chronic stimulation electrodes were stereotaxically implanted in and around the hypothalamus of 29 rats. Under awake conditions, 38 locomotor sites were tested in a runway apparatus. Behaviors occurring between the onset of stimulation and the first step were recorded on videotape. Points on the rat were digitized at sampling rate of 6 Hz to produce measures of head angles in the vertical, horizontal, and sagittal planes. The priming paradigm was used with a current selected for each site that was minimally sufficient to produce reliable stepping. In trials at approximately 1-min intervals, a 5-s train of stimulation (the control) was followed by a second train (the test) delivered 5–20 s later. Initiation latency on control trains was strongly correlated with head movement measures. Vertical and lateral head movements were independent of one another. Together, their frequency and extent accounted for 85% of the variance in locomotor initiation latencies. In effective priming trials, when locomotor initiation latencies were reduced on the test train, the frequency and extent of vertical and lateral head movements were also reduced. In non-effective priming trials, when latencies were not reduced, head movements were not reduced. Head scanning and locomotor initiation reflect reciprocal processes.

Locomotion and head scanning initiated by hypothalamic stimulation are inversely related.

Stimulation in the hypothalamus elicits locomotor stepping. Before stepping is initiated, head scanning movements occur. We determined the relationships between the latency of locomotor initiation and the number, extent and direction of the head scanning movements. Chronic stimulation electrodes were stereotaxically implanted in and around the hypothalamus of 29 rats. Under awake conditions, 38 locomotor sites were tested in a runway apparatus. Behaviors occurring between the onset of stimulation and the first step were recorded on videotape. Points on the rat were digitized at sampling rate of 6 Hz to produce measures of head angles in the vertical, horizontal, and sagittal planes. The priming paradigm was used with a current selected for each site that was minimally sufficient to produce reliable stepping. In trials at approximately 1-min intervals, a 5-s train of stimulation (the control) was followed by a second train (the test) delivered 5–20 s later. Initiation latency on control trains was strongly correlated with head movement measures. Vertical and lateral head movements were independent of one another. Together, their frequency and extent accounted for 85% of the variance in locomotor initiation latencies. In effective priming trials, when locomotor initiation latencies were reduced on the test train, the frequency and extent of vertical and lateral head movements were also reduced. In non-effective priming trials, when latencies were not reduced, head movements were not reduced. Head scanning and locomotor initiation reflect reciprocal processes. © 1999 Elsevier Science B.V. All rights reserved.

Microstimulation mapping of the basal forebrain in the anesthetized rat: The “preoptic locomotor region”

Previous studies have indicated that the basal forebrain at the level of the preoptic area contains neurons which participate in the initiation of locomotion. This study attempted to localize those neurons by mapping sites at which 25- and 50-μA stimulation (50 Hz, 0.5 ms cathodal pulses, 10-s trains) initiated hindlimb stepping. Anesthetized rats were held in a stereotaxic apparatus supported by a sling so that stepping movements rotated a wheel. Anesthesia was maintained by periodic injections of Nembutal (7 mg/kg) supplemented by lidocaine injections. Stimulation was applied through 50–70-μm diameter pipettes filled with 2 M NaCl at approximately 1600 sites in the basal forebrain, adjacent thalamus, and striatum. A circumscribed grouping of 25-μA locomotor sites, centered in the lateral preoptic area, defined the preoptic locomotor region. It extended into the ventral bed nucleus of the stria terminalis, the lateral part of the medial preoptic area, the anterior hypothalamic area, the medial and rostral parts of the ventral pallidum, medial substantia innominata, and the horizontal limb of the diagonal band. This general region is known to project to the midbrain locomotor region and the ventral tegmental area; it is proposed to initiate locomotion in service of primary motivational systems. Among the structures generally negative for locomotor sites were the dorsal and ventral striata, septal complex, bed nucleus of stria terminalis, and lateral ventral pallidum and substantia innominata. These findings indicate that low current stimulation applied to a circumscribed area centered in the lateral preoptic area produces locomotor stepping in the anesthetized rat. Whether the activated elements in this preoptic locomotor region are cells or fibers is not yet known. The degree of localization afforded by these findings indicates that the areas that are most likely to contain the mediating elements are quite limited in extent.

Locomotor stepping elicited by electrical stimulation of the hypothalamus persists after lesion of descending fibers of passage

Locomotion initiated by electrical stimulation of the lateral hypothalamus could be due to activation of local neurons or of fibers of passage descending from locomotor regions in the basal forebrain. This study mapped hypothalamic sites for electrically elicited locomotion in six rats with electrolytic lesions of the ipsilateral basal forebrain sources of descending fibers of passage. For mapping, anesthetized rats were held in a stereotaxic apparatus supported by a sling so that stepping movements rotated a wheel. Anesthesia was maintained by periodic injections of Nembutal (7 mg/kg) supplemented by lidocaine injections. Stimulation (25 and 50 μA, 50 Hz, 0.5 msec cathodal pulses, 10 sec trains) was applied through 50–80 μm diameter pipettes filled with 2 M saline. In all cases, locomotor stepping could be elicited by stimulation in sites ipsilateral to the lesion at currents of 50 μA or less. In the one case in which 25 μA sites were not found in the lateral hypothalamus, the lesion extended caudally to within 1 mm of the stimulation sites. These findings do not exclude a locomotor role for fibers of passage but they suggest that activation of lateral hypothalamic neurons is sufficient to initiate locomotion.

Initiation and execution of locomotion elicited by diencephalic stimulation: Regional differences in response to Nembutal

At moderate levels of Nembutal, within the anesthetic range, locomotor stepping can be elicited by brain stimulation. We determined if Nembutal (7, 14 and 28 mg/kg) had different effects on locomotion elicited by stimulation at different brain regions. Two regions were compared: the medial forebrain bundle (MFB, 13 sites) and the areas medial and dorsal to it (MED/DORSAL, 20 sites). Locomotion was produced by electrical stimulation (50 μA, 0.5 msec pulses, 10 to 160 Hz) of unrestrained rats in a rotary runway. The latency to initiate locomotion and the time to complete 1 revolution of the rotary were measured. With no drug, MFB locomotion was initiated sooner but took longer to complete than MED/DORSAL locomotion. Nembutal at 7 mg/kg did not affect initiation of MFB or MED/DORSAL locomotion. Nembutal at 14 mg/kg shortened MFB initiations, but this dose prolonged MED/DORSAL initiations. Initiations with both types of sites were blocked with 28 mg/kg. The 7 and 14 mg/kg doses prolonged the locomotor completion times of the MFB sites but not of the MED/DORSAL sites. The results indicate that the response to Nembutal differs qualitatively for locomotion elicited by stimulation of the MFB and locomotion elicited by stimulation of the medial and dorsal hypothalamus. The mechanisms underlying the difference remain to be elucidated; they may relate to nonlocomotor behaviors also elicited by stimulation or to the motivational states reflected in those behaviors.

Latency to initiate locomotion elicited by stimulation of the diencephalon positively correlates in awake and anesthetized rats

Locomotor stepping can be elicited by brain stimulation at various diencephalic sites under moderate levels of Nembutal. This study determined if locomotor initiation measured under anesthesia provides a valid measure of the intersite factors which determine initiation in the awake condition. We compared the latencies to inititate locomotor stepping elicited by electrical stimulation (50 μA, 0.5-msec pulses, 10 to 160 Hz) by rats tested while awake and unrestrained in a rotary runway or anesthetized and held in a stereotaxic apparatus. In the latter tests, initial anesthesia was provided by Nembutal (25 mg/kg) and 2% halothane and maintenace anesthesia was provided by 7 mg/kg as needed and local injections of lidocaine. For 30 sites in 16 rats, average locomotor initiation latency in the awake condition and the shortest latencies in the anesthetized condition were positively correlated (r=.78). Locomotion at sites with long latencies in the awake condition was frequently blocked in the anesthetized condition, but sites with short latencies were rarely blocked. The results indicate that the shortest locomotor latencies in the anesthetized condition approximate the latencies measured in the awake condition. It is concluded that the anesthetized condition can provide valid initiation measures, but sites with long latencies in the awake condition are prone to depression under anesthesia.

Locomotor stepping initiated by glutamate injections into the hypothalamus of the anesthetized rat.

Glutamate (50 mM, 50 nl) injected into the tuberal and posterior hypothalamus was tested for capacity to elicit locomotor stepping. Rats (n = 23) were anesthetized with Nembutal and suspended by a sling in a stereotaxic apparatus such that locomotor stepping rotated a wheel. In 61 of 275 sites tested, stepping was initiated by glutamate injections within 60 s. Positive sites were widespread and contained in the lateral hypothalamus, the perifornical area, the dorsomedial nucleus, and the zona incerta. The perifornical and lateral hypothalamic sites were most likely to have locomotor responses and the shortest latencies. These findings indicate that selective activation of hypothalamic neurons as opposed to fibers of passage can initiate locomotion.

Midbrain stimulation in the anesthetized rat: Direct locomotor effects and modulation of locomotion produced by hypothalamic stimulation

The midbrain contains circuits that modulate locomotion. To delineate some of the involved regions, low-level stimulation (25 μA, 10 s train of 0.5 ms pulses at 50 Hz) was applied to the midbrain during locomotor stepping. Stepping was elicited in the anesthetized (pentobarbital, 40 mg/kg) rat by stimulating the hypothalamus with 0.5 ms pulses at 40 Hz at various currents. The rat was held in a stereotaxic apparatus such that locomotor stepping movements turned a wheel. Facilitation of locomotion was produced by stimulation in the anterior ventromedial midbrain and in the posterodorsal midbrain. When presented alone, such stimulation produced locomotion. Inhibition of locomotion was produced by stimulation of the superior colliculus (ventral layers) and the ventromedial midbrain. Additional inhibitory sites were found in the central gray and the lateral tegmentum. Inhibitory collicular stimulation, when presented alone, was characterized by the absence of any hindlimb response. Inhibitory ventromedial stimulation, when presented alone, frequently produced poststimulation locomotion and when presented with hypothalamic stimulation was characterized by postinhibitory increases in locomotion. These results indicate that: (1) the locomotor effects of stimulation in midbrain and hypothalamic sites can summate; (2) multiple locomotor suppressive systems are present in the midbrain and among them are a collicular system and a ventromedial system.

Tactile stimulation inhibits locomotion produced by stimulation of the lateral hypothalamus in the anesthetized rat

Rats were anesthetized with Nembutal and suspended in a stereotaxic apparatus so that their limbs contacted the outer surface of a 30-cm diameter wheel. Stimulation in the lateral hypothalamus (0.5 msec cathodal pulses at 50 Hz, up to 150 μA) produced locomotor stepping which turned the wheel. During continuous locomotion maintained by electrical stimulation, tactile stimulation was applied manually for 5-sec periods to various body regions in six subjects. Profound inhibition of locomotor stepping was produced in all subjects by mild pressure applied to the nose or the vibrissae pad. Vibrissae pad stimulation either ipsilateral or contralateral to the stimulation site was effective. Stroking of the vibrissae inhibited locomotion in only 2 of 6 subjects and in these cases the effective stimulation was contralateral to the brain stimulation site. With pressure applied to more caudal body regions, including the dorsum, the abdomen and the tail, the inhibition was less pronounced and less consistent than that produced by pressure on the snout region. The findings indicate that the tactile system provides an important modulation of locomotion and that the anesthetized locomoting rat offers a useful preparation for its further study.