From cineradiography to biorobots: an approach for designing robots to emulate and study animal locomotion.

Robots are increasingly used as scientific tools to investigate animal locomotion. However, designing a robot that properly emulates the kinematic and dynamic properties of an animal is difficult because of the complexity of musculoskeletal systems and the limitations of current robotics technology. Here, we propose a design process that combines high-speed cineradiography, optimization, dynamic scaling, three-dimensional printing, high-end servomotors and a tailored dry-suit to construct Pleurobot: a salamander-like robot that closely mimics its biological counterpart, Pleurodeles waltl Our previous robots helped us test and confirm hypotheses on the interaction between the locomotor neuronal networks of the limbs and the spine to generate basic swimming and walking gaits. With Pleurobot, we demonstrate a design process that will enable studies of richer motor skills in salamanders. In particular, we are interested in how these richer motor skills can be obtained by extending our spinal cord models with the addition of more descending pathways and more detailed limb central pattern generator networks. Pleurobot is a dynamically scaled amphibious salamander robot with a large number of actuated degrees of freedom (DOFs: 27 in total). Because of our design process, the robot can capture most of the animal's DOFs and range of motion, especially at the limbs. We demonstrate the robot's abilities by imposing raw kinematic data, extracted from X-ray videos, to the robot's joints for basic locomotor behaviours in water and on land. The robot closely matches the behaviour of the animal in terms of relative forward speeds and lateral displacements. Ground reaction forces during walking also resemble those of the animal. Based on our results, we anticipate that future studies on richer motor skills in salamanders will highly benefit from Pleurobot's design.

Flexibility of the axial central pattern generator network for locomotion in the salamander.

In tetrapods, limb and axial movements are coordinated during locomotion. It is well established that inter- and intralimb coordination show considerable variations during ongoing locomotion. Much less is known about the flexibility of the axial musculoskeletal system during locomotion and the neural mechanisms involved. Here we examined this issue in the salamander Pleurodeles waltlii, which is capable of locomotion in both aquatic and terrestrial environments. Kinematics of the trunk and electromyograms from the mid-trunk epaxial myotomes were recorded during four locomotor behaviors in freely moving animals. A similar approach was used during rhythmic struggling movements since this would give some insight into the flexibility of the axial motor system. Our results show that each of the forms of locomotion and the struggling behavior is characterized by a distinct combination of mid-trunk motor patterns and cycle durations. Using in vitro electrophysiological recordings in isolated spinal cords, we observed that the spinal networks activated with bath-applied N-methyl-d-aspartate could generate these axial motor patterns. In these isolated spinal cord preparations, the limb motor nerve activities were coordinated with each mid-trunk motor pattern. Furthermore, isolated mid-trunk spinal cords and hemicords could generate the mid-trunk motor patterns. This indicates that each side of the cord comprises a network able to generate coordinated axial motor activity. The roles of descending and sensory inputs in the behavior-related changes in axial motor coordination are discussed.

Fictive rhythmic motor patterns produced by the tail spinal cord in salamanders.

Most investigations into the role of the body axis in vertebrate locomotion have focused on the trunk, although in most tetrapods, the tail also plays an active role. In salamanders, the tail contributes to propulsion during swimming and to dynamic balance and maneuverability during terrestrial locomotion. The aim of the present study was to obtain information concerning the neural mechanisms that produce tail muscle contractions during locomotion in the salamander Pleurodeles waltlii. We recorded the ventral root activities in in vitro spinal cord preparations in which locomotor-like activity was induced via bath application of N-methyl-d-aspartate (20µM) and d-serine (10µM). Recordings showed that the tail spinal cord is capable of producing propagated waves of motor activity that alternate between the left and right sides. Lesion experiments further revealed that the tail rhythmogenic network is composed of a double chain of identical hemisegmental oscillators. Finally, using spinal cord preparations bathed in a chamber partitioned into two pools, we revealed efficient short-distance coupling between the trunk and tail networks. Together, our results demonstrate the existence of a pattern generator for rhythmic tail movements in the salamander and show that the global architecture of the tail network is similar to that previously proposed for the mid-trunk locomotor network in the salamander. Our findings further support the view that salamanders can control their trunk and tail independently during stepping movements. The relevance of our results in relation to the generation of tail muscle contractions in freely moving salamanders is discussed.

Effects of locomotor training on hindlimb regeneration in the urodele amphibian Pleurodeles waltlii.

1. The effects of locomotor training on hindlimb regeneration were studied in the urodele amphibian Pleurodeles waltlii. 2. After amputation of one hindlimb at mid-femur, adult animals were subjected to regular training sessions (1 h daily, 5 days a week, over 8 months) of terrestrial stepping. 3. Eight months post-amputation, trained animals exhibited regenerated limbs of reduced size as compared to animals kept in their aquaria. Histological data showed an abnormal regeneration of both the femur and distal structures (e.g. digit muscles, metatarsi and phalanges) while medial structures (e.g. tibia and fibula) were totally re-formed. The study of the electromyographical activity in regenerated limbs during stepping and that of their reflex responsiveness to electrical stimulation showed that both motor and sensory innervations were functional in the limb stump of trained animals. 4. The regenerative capacity of the abnormal stumps was preserved since following a second amputation a quite normal hindlimb was regenerated in 3 months, provided the re-amputated animals were not trained to terrestrial stepping. 5. The stress due to handling, change in locomotor medium (aquatic vs. terrestrial) and the friction of the wound epidermis with the ground were not involved in the disruption of limb regeneration. 6. The locomotor pattern, the reflex responsiveness and the muscle fibre composition were similar in supernumerary forelimbs grafted on the back and in normal forelimbs. However, the supernumerary forelimbs regenerated normally even in animals subjected to locomotor training while the hindlimb did not. It is concluded that the disrupting effects of locomotor training on limb regeneration were localized to the the limb directly involved in locomotion. 7. The mechanisms underlying abnormal limb regeneration in animals subjected to locomotor training are discussed.

Serotonergic systems in the spinal cord of the amphibian urodele Pleurodeles waltl.

The role of the monoamine serotonin (5-HT) in modulating the neural networks underlying axial locomotor movements was studied in an adult amphibian urodele, Pleurodeles waltl. 5-HT was applied to an in vitro brainstem-spinal cord preparation of P. waltl, which displayed fictive axial locomotor patterns following bath application of N-methyl-D-aspartate (5 microM) with D-serine (10 microM). Our results showed that 5-HT (1-25 microM) produces a reversible increase in the cycle duration and the duration of rhythmic bursting activity recorded extracellularly from ventral roots innervating the axial musculature. When applied alone, 5-HT does not trigger axial locomotor activity. The distribution pattern of 5-HT immunoreactive (5-HT-ir) cells along the spinal cord was investigated both in intact and in chronic spinal animals. The number of 5-HT-ir cell bodies is higher at brachial levels and decreases through crural levels. Sparse oval or fusiform 5-HT-ir somata are present within the gray matter, just ventrolateral to the central canal. Longitudinal fibers were detected throughout the entire white matter, except in the medial part of the dorsal funiculi. Two columns of intensely labeled and profusely branching thick and thin fibers associated with numerous varicosities run continuously along the ventrolateral surface of the spinal cord. Three weeks following full spinal cord transection at the level of the second spinal root, all longitudinal processes had disappeared, indicating their supraspinal origin, whereas the ventrolateral plexes remained, suggesting that they originated from intraspinal 5-HT-ir cell bodies. Our data showing that spinal 5-HT is organized according to a rostrocaudal gradient suggest that the 5-HT systems of P. waltl are not related to the presence of limb motor pools but more likely are related to axial central pattern generators (CPGs) networks down the length of the spinal cord. The possible involvement of these two sources (descending vs. intraspinal) of 5-HT innervation in the modulation of the axial CPGs is discussed.

Fictive rhythmic motor patterns induced by NMDA in an in vitro brain stem-spinal cord preparation from an adult urodele.

An in vitro brain stem-spinal cord preparation from an adult urodele (Pleurodeles waltl) was developed in which two fictive rhythmic motor patterns were evoked by bath application of N-methyl-D-aspartate (NMDA; 2.5-10 microM) with D-serine (10 microM). Both motor patterns displayed left-right alternation. The first pattern was characterized by cycle periods ranging between 2.4 and 9. 0 s (4.9 +/- 1.2 s, mean +/- SD) and a rostrocaudal propagation of the activity in consecutive ventral roots. The second pattern displayed longer cycle periods (8.1-28.3 s; 14.2 +/- 3.6 s) with a caudorostral propagation. The two patterns were inducible after a spinal transection at the first segment. Preliminary experiments on small pieces of spinal cord further suggested that the ability for rhythm generation is distributed along the spinal cord of this preparation. This study shows that the in vitro brain stem-spinal cord preparation from Pleurodeles waltl may be a useful model to study the mechanisms underlying the different axial motor patterns and the flexibility of the neural networks involved.

Epaxial and limb muscle activity during swimming and terrestrial stepping in the adult newt, Pleurodeles waltl.

We have investigated the patterns of activation of epaxial musculature during both swimming and overground stepping in an adult newt (Pleurodeles waltl) with the use of electromyographic (EMG) recordings from different sites of the myomeric muscle dorsalis trunci along the body axis. The locomotor patterns of some limb muscles have also been investigated. During swimming, the epaxial myomeres are rhythmically active, with a strict alternation between opposite myomeres located at the same longitudinal site. The pattern of intersegmental coordination consists of three successively initiated waves of EMG activity passing posteriorly along the anterior trunk, the midtrunk, and the posterior trunk, respectively. Swimming is also characterized by a tonic activation of forelimb (dorsalis scapulae and extensor ulnae) and hindlimb (puboischiotibialis and puboischiofemoralis internus) muscles and a rhythmic activation of muscles (latissimus dorsi and caudofemoralis) acting both on limb and body axis. The latter matched the activation pattern of epaxial myomeres at the similar vertebral level. During overground stepping, the midtrunk myomeres express single synchronous bursts whereas the myomeres of the anterior trunk and those of the posterior trunk display a double bursting pattern in the form of two waves of EMG activity propagating in opposite directions. During overground stepping, the limb muscles and muscles acting on both limb and body axis were found to be rhythmically active and usually displayed a double bursting pattern. The main conclusion of this investigation is that the patterns of intersegmental coordination during both swimming and overground stepping in the adult newt are related to the presence of limbs and that they can be considered as hybrid lampreylike patterns. Thus it is hypothesized that, in newt, a chain of coupled segmental oscillatory networks, similar to that which constitutes the central pattern generator (CPG) for swimming in the lamprey, can account for both trunk motor patterns if it is influenced by limb CPGs in a way depending on the locomotor mode. During swimming, the segmental networks located close to the girdles receive extra tonic excitation coming from the limb CPGs, whereas during stepping, the axial CPGs are entrained to some extent by the limb oscillators.

Fictive motor activities in adult chronic spinal rats transplanted with embryonic brainstem neurons.

The present study was designed to examine the effects of an intraspinal transplantation of embryonic brainstem neurons on fictive motor patterns which can develop in hindlimb nerves of adult chronic spinal rats. Seventeen adult rats were spinalized at T8-9 level and, 8 days later, a suspension of embryonic cells obtained either from the raphe region (RR, n = 8) or from the locus coeruleus (LC, n = 9) was injected caudally (T12-13) to the cord transection. Eight control animals (control rats) were spinalized and injected with vehicle under the same conditions. One to three months later, the animals were decorticated and fictive motor patterns were recorded in representative hindlimb nerves. The data revealed that both control and grafted spinal rats could exhibit two distinctly different fictive motor patterns, one which could be associated with stepping and the other with hindlimb paw shaking. They further showed that following transplantation of embryonic RR or LC neurons the excitability of the spinal stepping generator was increased, whereas that of the spinal neural circuits which generate hindlimb paw shaking was not significantly affected. A histological analysis performed on the spinal cord segments below the transection revealed complete absence of serotonin and noradrenaline immunoreactivity in control spinal animals and, in both types of grafted rats, an extensive monoaminergic reinnervation with synaptic contacts between monoaminergic transplanted neurons and host interneurons and/or motoneurons. The possible mechanisms by which grafted monoaminergic neurons can influence the spinal motor networks are discussed.

Fictive locomotion in the adult thalamic rat.

In immobilized adult thalamic rats, electrical stimulation of sites within the lateral hypothalamic area (LHA) or the mesencephalic locomotor region (MLR) were found to elicit fictive locomotor patterns in hindlimb muscle nerves. Significant differences were found between several characteristics (average cycle period, locomotor episode duration, intralimb and interlimb coordination patterns) of the LHA-induced and MLR-induced fictive locomotor activities. These findings support the hypothesis that LHA and MLR play different functional roles during locomotion.

An Electromyographic Study of the Hindlimb Locomotor Movements in the Acute Thalamic Rat.

In this paper we have analysed the patterns of muscular activities that underlie hindlimb locomotor movements in the acute thalamic rat. Electromyographic activities of muscles representative of the functional muscle groups of the hindlimbs were recorded bipolarly during locomotion in acute thalamic rats. Locomotor movements occurred spontaneously, but could also be induced by electrical stimulation (0.1 ms pulses; 30 - 70 Hz; 75 - 300 microA) of the lateral hypothalamic area. The two hindlimbs displayed a wide variety of coordination patterns during both types of locomotion. However, alternated coordination of the hindlimbs occurred more frequently during induced than during spontaneous locomotion. Correspondingly, the duration of the spontaneous step cycles had a tendency to be shorter than that of the evoked step cycles, although they had a quite similar range. The patterns of muscular activities within one hindlimb were similar during spontaneous and induced locomotion. During each step cycle, (i) the hip and ankle flexors usually displayed a single burst in alternation with that displayed by the hip, knee and ankle extensors, (ii) a double bursting pattern was sometimes observed in flexors during fast locomotor movements, (iii) within flexors and extensors, muscles were recruited sequentially, and (iv) the activation of other muscles (semitendinosus, rectus femoris, extensor and flexor digitorum longus) consisted of single or double bursting patterns.

An intracellular study of muscle primary afferents during fictive locomotion in the cat.

1. Presynaptic activity of identified primary afferents from flexor, extensor, and bifunctional hindlimb muscles was studied with intra-axonal recordings during fictive locomotion. Fictive locomotion appeared spontaneously in decorticate cats (n = 9), with stimulation of the mesencephalic locomotor region (n = 4), and in spinal cats injected with clonidine or nialamide and L-DOPA (n = 4). Representative flexor and extensor muscle nerves, recorded to monitor the locomotor pattern and dorsal rootlets of the sixth and seventh lumbar segments, were recorded simultaneously to monitor dorsal root potentials (DRPs). 2. From responses to muscle stretches and, in some instances, twitch contractions of the parent muscle, 75% of the single units examined were putatively identified as spindle afferents (40/53). On the basis of conduction velocity and stimulation threshold, 73% of these were further classified as group I fibers (29/40), the rest as group II fibers. 3. All units (n = 53 with resting potential more negative than -45 mV) showed fluctuations of their membrane potential (up to 1.5 mV) at the rhythm of the fictive locomotion. Subsequent averaging of these fluctuations over several cycles revealed that 89% of all units displayed a predominant wave of depolarization during the flexor phase, followed by a trough of repolarization. In 79% of the units, there was also a second, usually smaller, depolarization during the extensor phase. The relative size of each wave of depolarization could vary with different episodes of fictive locomotion in the same unit and among various afferents from the same muscle in the same experiment. 4. The firing frequency of some afferents from the ankle flexor tibialis anterior (5/16) and the bifunctional muscle posterior biceps-semitendinosus (4/15) was phasically modulated along the fictive step cycle. The maximum frequency always occurred during the flexor phase, i.e., during the largest depolarization of the unit. Because of the absence of phasic sensory input in the curarized animal, we assume that the phasic discharges were generated within the spinal cord and antidromically propagated. Phasic firing was never encountered in afferents from extensor muscles such as triceps surae (0/15) and vastus lateralis (0/4). 5. The results demonstrate that the pattern of rhythmic depolarization accompanying fictive locomotion is similar for the majority of flexor, extensor, and bifunctional group I (and possibly group II) muscle spindle primary afferents. They further indicate that there is a specific phasic modulation of antidromic firing for some flexor and bifunctional muscle spindle afferents.(ABSTRACT TRUNCATED AT 400 WORDS)

Phase-dependent modulation of primary afferent depolarization in single cutaneous primary afferents evoked by peripheral stimulation during fictive locomotion in the cat.

Previous results from our laboratory have shown with intra-axonal recordings that hindfoot cutaneous primary afferents are subjected to rhythmic depolarizations during fictive locomotion (L-PAD) suggesting that cutaneous presynaptic mechanisms are activated by the central locomotor program. In this study, we examined the transmission in pathways responsible for primary afferent depolarizations (PAD) of cutaneous fibres during spontaneous fictive locomotion in decorticate cats and in spinal cats injected with nialamide and L-DOPA. PADs were evoked (E-PADs) by electrical stimulation of peripheral nerves and recorded intra-axonally with micropipettes in identified superficialis peroneal (SP; n = 7) and tibialis posterior (TP; n = 17) cutaneous primary afferents. Results showed that the amplitude of E-PADs, which were superimposed on the L-PAD, was deeply modulated throughout the locomotor cycle; decreasing to reach a minimum during the flexor phase and increasing to a maximum during the extensor phase. The results were not statistically different in fibres of the two nerves and in both types of preparation. The amplitude of E-PADs was always maximum during the extensor phase whether there was a large L-PAD or not during that phase. This suggests that the presynaptic mechanisms activated by central locomotor networks (L-PAD) and those activated by peripheral inputs (E-PAD) may in part be controlled differently. The results thus show that the transmission in PAD pathways activated by cutaneous inputs is phasically modulated by the central pattern generator for locomotion. This strongly suggests that the presynaptic inhibition in cutaneous fibres evoked by the movement-related feedback during real locomotion could be similarly modulated.

Interlimb coordination during fictive locomotion in the thalamic cat.

Efferent discharges in muscle nerves of the four limbs were recorded simultaneously during spontaneous fictive locomotion in thalamic cats with the goal of understanding how the central nervous system controls interlimb coordination during stepping. The onset of the bursts of activity in the nerve of a selected flexor muscle in each limb allowed the temporal and the phase relationships between the fictive step cycle of a pair of limbs to be determined. Our main results are the following: 1) the fictive step cycles of the two forelimbs are always strictly alternated whereas the phasing of the step cycles of either the two hindlimbs or pairs of homolateral or diagonal limbs is more variable; 2) the time interval between the onsets of the flexor bursts of one of the two pairs of diagonal limbs is independent of the step cycle duration; 3) distinct patterns of interlimb coordination exist during fictive locomotion; a small number of patterns of coordination involving all four limbs, which correspond to the walking and the trotting gaits in the intact cat, occur very frequently. The results demonstrate that the central nervous system deprived of phasic afferent inputs from the periphery has the capacity to generate most of the patterns of interlimb coordination which occur during real locomotion. They further support the view that the central pattern of interlimb coordination essentially results from diagonal interaction between a forelimb generator for locomotion and a hindlimb one.

Intra-axonal recordings of cutaneous primary afferents during fictive locomotion in the cat.

1. Cutaneous primary afferents were recorded intracellularly during fictive locomotion in decorticated cats with the goal of improving our understanding of how locomotor networks might centrally control the transmission in cutaneous pathways at a presynaptic level. 2. Identified cutaneous axons from superficialis peroneal nerve (SP) or tibialis posterior nerve (TP) were recorded intracellularly together with the electroneurograms (ENGs) of representative flexor and extensor muscle nerves of the hindlimb as well as dorsal root potential from L6 or L7 (DRP). Fictive locomotion occurred spontaneously after decortication (n = 12) or was induced by stimulation of the mesencephalic locomotor region (MLR) (n = 6). 3. The results revealed that all cutaneous axons (82 units with resting potential greater than 45 mV) showed fluctuations of their membrane potential (greater than or equal to 0.5 mV) at the rhythm of the fictive locomotion. The characteristics of fluctuation patterns, common to all cutaneous units, consisted of two depolarization waves per cycle: one related to the flexor activity, the other related to the extensor activity. The flexor-related depolarization was followed by a sharp trough of membrane repolarization. The extensor-related depolarization usually overlapped partly with the flexor-depolarization of the following cycle. The relative size of each depolarization could vary among different afferents of the same nerve in the same animal. Hence, maximal depolarization could occur in different parts of the locomotor cycle, but, for the majority of units (82%), it occurred during the flexor activity. These results were similar for SP and TP units. 4. Twenty percent of the units were discharging with a constant or irregular frequency. Phasic antidromic discharges related to locomotor ENGs were rarely encountered (5/82 units). 5. Linear regression analysis of the temporal relationships between fluctuations of membrane potential of cutaneous axons and locomotor bursts over several cycles showed that the timing of presynaptic events in cutaneous afferents is related to the events of the locomotor output. However, the same type of analysis showed that the amplitude of axonal depolarizations and the amplitude of flexor and extensor locomotor bursts could vary independently. Tight temporal relationships were also found between the depolarizations recorded in cutaneous units and the fluctuations recorded at the dorsal root level (DRP). 6. Based on the assumption that the locomotor fluctuations of cutaneous membrane potential are mediated through the primary afferent depolarization (PAD) pathways associated with presynaptic inhibition, it is proposed that the central pattern generator for locomotion (CPG) could phasically control the efficacy of transmission of cutaneous pathways at a presynaptic level as part of the locomotor program.

Discharge patterns of gamma motoneurone populations of extensor and flexor hindlimb muscles during walking in the thalamic cat.

Two monopolar recordings of the whole activity in a fine nerve branch innervating the gastrocnemius lateralis (GL) or the sartorius medialis (SM) muscle were obtained during spontaneous walking in thalamic cats. Using a special electronic device, the potentials of several groups of efferent (alpha and gamma) and afferent (I and II) fibres constituting the whole nerve activity were separated. In the present paper we compare the data obtained for an ankle extensor (GL) and a hip-knee flexor (SM) during the step cycle. In both muscles the gamma motoneurone population is activated in parallel with the alpha motoneurone population. Usually, between the cyclic locomotor discharges, the GL gamma neurones are tonically active whereas the SM gamma neurones are silent. During muscle contraction, the group I and II afferent discharges are both length and gamma dependent, but the prevailing factor is the muscle shortening for the GL afferents and the cyclic gamma drive for the SM afferents. Both dynamic and static fusimotor efferents appear to be activated during muscle contraction, but on indirect evidence it is suggested that dynamic action prevails in GL spindles whereas static action dominates in SM spindles.

Rhythmic fluctuations of dorsal root potentials and antidromic discharges of primary afferents during fictive locomotion in the cat.

1. This study examines rhythmical activity of primary afferents occurring during "fictive" locomotion in decorticate paralyzed cats. Oscillations of the dorsal root potential (DRP) at the frequency of the locomotor rhythm have been observed at the lumbosacral and cervical levels. In addition, rhythmic antidromic discharges of primary afferent units have been recorded from the proximal stumps of cut dorsal root filaments. A detailed study of the relationships between the DRP fluctuations, the antidromic discharges, and the locomotor activity monitored by recording extensor and flexor muscle nerves is presented. 2. Typical DRP recordings from both lumbosacral and cervical levels show two negative waves (N1 and N2) separated by positive troughs (P1 and P2) in each locomotor cycle. Linear regression analyses indicate that the first negative wave (which generally has the largest amplitude) is related to the flexor activity whereas the second is related to the extensor activity. The relative amplitude of the two negative waves may vary without apparent concomitant changes in the recorded flexor or extensor motor nerves. The positive troughs occur respectively close to the period of transition between flexor and extensor activities and between extensor and flexor activities. 3. DRPs of similar period and amplitude can be observed in different ipsilateral roots recorded simultaneously. The DRPs recorded bilaterally from the same segment have the same periodicity but are out-of-phase. Point-to-point variations of amplitude in bilaterally recorded roots are not correlated. This suggests that the polarization of primary afferents on one side is mainly related to the locomotor events on that side. DRPs have been recorded in cats spinalized at Th13 and injected with nialamide and l-DOPA. This suggests that although the supraspinal contribution may be important, at least part of the DRPs may result from locomotor activity within the spinal cord itself. 4. A salient finding in our experiments was that of rhythmic antidromic unit discharges in the proximal stump of cut dorsal root filaments. Of the 194 units recorded, 19% (37/194) discharged in distinct bursts occurring at fixed times in the locomotor cycle. The majority of the units discharged either one burst during the period of flexor or extensor activity or one burst during one of the two periods of transition. Three units discharged two bursts per locomotor cycle. The frequency of the antidromic discharges of some units in one limb were also found to be modulated by stimulation of the skin or passive manipulation of the limbs.(ABSTRACT TRUNCATED AT 400 WORDS)

Comparison between ventral spinocerebellar and rubrospinal activities during locomotion in the cat.

During fictive locomotion of the thalamic cat, rhythmic activity related to the efferent discharges in hindlimb nerves was found in rubrospinal neurons (Arshavsky et al., this issue). Since the movements were abolished by curarization, this modulation could not result from rhythmic peripheral inputs and had therefore a central origin. Taking into account the existence of spinal generators, it was suggested that ascending pathways transmit rhythmic activity from these spinal centers to the supraspinal ones. Preliminary results have been obtained for neurons of the ventral spinocerebellar tract (VSCT) recorded during fictive locomotion: (1) their discharge is rhythmically modulated at the periodicity of the locomotor rhythm; (2) their discharge pattern can be complex and variable in relation with the complexity and variability of the pattern of efferent activity in various muscle nerves of the ipsilateral hindlimb; (3) their responses to phasic afferent stimulation of the ipsilateral hindlimb are modulated in parallel with their locomotor-related activity. These results show that VSCT neurons convey information on the central spinal activity during locomotion, and suggest that these neurons contribute to the activity of lumbar-projecting rubrospinal neurons which have similar characteristics.

Efferent and afferent activity in a gastrocnemius nerve branch during locomotion in the thalamic cat.

The firing patterns of alpha and gamma efferent fibres and of group I and group II afferent fibres innervating the gastrocnemius muscle were observed during spontaneous locomotor movements in the thalamic cat. Multi-unit discharges of each kind of fibre were obtained by electronic sorting of the action potentials from the whole activity of a thin branch of gastrocnemius lateralis or medialis nerve. The main results were: During the locomotor cycle the activity of the afferent and efferent populations was highly modulated. alpha- and gamma-motoneurones were co-activated within the locomotor cycle during ankle plantar-flexion. The gamma discharge began to rise earlier and to fall later than did the alpha discharge. The amplitude of the gamma discharge, unlike that of the alpha discharge, was largely independent of the vigour of walking. Between the cyclic discharges, most of gamma populations were tonically active whereas alpha populations were silent. Subgroups of the alpha and gamma populations were not usually activated according to the cell-size principle, but, the activation of the latest gamma subgroup always preceded that of the earliest alpha subgroup. Modulation of the group I and II afferent discharges was closely related to the cyclic length changes of the parent muscle. Fusimotor activation during the active shortening of gastrocnemius muscle prevented the afferent discharges from pausing. The pattern of afferent and efferent activity during selective curarisation of the extrafusal junctions indicated that the discharge of static gamma-motoneurones is modulated during the locomotor cycle. After curarisation of both extrafusal and intrafusal junctions, an efferent-discharge pattern of central origin persisted alternately in extensor- and flexor-muscle nerves (fictive locomotion). The durations of the fictive locomotor cycle and of the cyclic discharge in the sartorius nerve were increased as a consequence of the suppression of phasic afferent inputs to the C.N.S. Maintained ankle dorsi-flexion slowed the fictive locomotor rhythm and elicited opposite effects, respectively excitation and depression, on the magnitude of the alpha and gamma discharges. Maintained ankle plantar-flexion scarcely perturbed the duration of the fictive locomotor cycle, but the duration of the sartorius-nerve discharge lengthened at the expense of that of the gastrocnemius discharge. Both gastrocnemius alpha- and gamma-motoneurones were depressed, the former considerably more than the latter. The roles of the gastrocnemius afferents and gamma-efferents during the locomotor cycle are discussed in the light of these results.

Evidence of rhythmic inhibitory synaptic influences in hindlimb motoneurons during fictive locomotion in the thalamic cat.

Intracellular recordings of various motoneurons of proximal hindlimb muscles were performed on thalamic paralyzed cats, during fictive locomotion that was either spontaneous or evoked by stimulation of the subthalamic region. In motoneurons innervating sartorius (medialis and lateralis), vasti (intermedius, medialis and lateralis) and anterior biceps-semimembranous, one depolarization occurred in each locomotor cycle, alternating with a phase of repolarization that was synchronous with the activation of the antagonistic muscle nerve. This latter phase could be decreased or reversed by intracellular injection of chloride ions or current, revealing the presence of inhibitory inputs onto motoneurons. The pattern of membrane potential variations was more complex in motoneurons of rectus femoris and posterior biceps-semitendinosus muscles, but phases of chloride dependent inhibition were nevertheless identified, mainly during the sartorius nerve activation in the case of rectus femoris, and during the vasti and anterior biceps-semimembranosus nerve activations in the case of posterior biceps-semitendinosus. These inhibitory influences were shown to be controlled by the level of activity in exteroceptive afferents. The characteristics of the excitatory and inhibitory inputs to the hindlimb motoneurons identified here are discussed in relation with the organization of the central pattern generator for locomotion.

Rhythmic antidromic discharges of single primary afferents recorded in cut dorsal root filaments during locomotion in the cat.

Single units recorded in the proximal stump of cut dorsal root filaments were found to antidromically discharge rhythmically during fictive locomotion in decorticate and paralyzed cats. Some units fired throughout the period of flexor or extensor nerve activity, whereas other units discharged near the transitional phases. Similar findings were made in acutely spinalized and paralyzed cats injected with L-DOPA, as well as in non-paralyzed decorticate cats walking on a treadmill. These results suggest that different types of primary afferents may be depolarized cyclically at different specific time in the step cycle by the central pattern generator for locomotion, and that this central control of the primary afferents may be involved in the modulation of the reflex transmission observed during locomotion.