Invasion of lesion territory by regenerating fibers after spinal cord injury in adult macaque monkeys.

In adult macaque monkeys subjected to an incomplete spinal cord injury (SCI), corticospinal (CS) fibers are rarely observed to grow in the lesion territory. This situation is little affected by the application of an anti-Nogo-A antibody which otherwise fosters the growth of CS fibers rostrally and caudally to the lesion. However, when using the Sternberger monoclonal-incorporated antibody 32 (SMI-32), a marker detecting a non-phosphorylated neurofilament epitope, numerous SMI-32-positive (+) fibers were observed in the spinal lesion territory of 18 adult macaque monkeys; eight of these animals had received a control antibody infusion intrathecally for 1 month after the injury, five animals an anti-Nogo-A antibody, and five animals received an anti-Nogo-A antibody together with brain-derived neurotrophic factor (BDNF). These fibers occupied the whole dorso-ventral axis of the lesion site with a tendency to accumulate on the ventral side, and their trajectories were erratic. Most of these fibers (about 87%) were larger than 1.3 µm and densely SMI-32 (+) stained. In the undamaged spinal tissue, motoneurons form the only large population of SMI-32 (+) neurons which are densely stained and have large diameter axons. These data therefore suggest that a sizeable proportion of the fibers seen in the lesion territory originate from motoneurons, although fibers of other origins could also contribute. Neither the presence of the antibody neutralizing Nogo-A alone, nor the presence of the antibody neutralizing Nogo-A combined with BDNF influenced the number or the length of the SMI-32 (+) fibers in the spinal lesion area. In summary, our data show that after a spinal cord lesion in adult monkeys, the lesion site is colonized by fibers, a large portion of which presumably originate from motoneurons.

Fate of rubrospinal neurons after unilateral section of the cervical spinal cord in adult macaque monkeys: effects of an antibody treatment neutralizing Nogo-A.

The present study describes in primates the effects of a spinal cord injury on the number and size of the neurons in the magnocellular part of the red nucleus (RNm), the origin of the rubrospinal tract, and evaluates whether a neutralization of Nogo-A reduces the lesioned-induced degenerative processes observed in RNm. Two groups of monkeys were subjected to unilateral section of the spinal cord affecting the rubrospinal tract; one group was subsequently treated with an antibody neutralizing Nogo-A; the second group received a control antibody. Intact animals were also included in the study. Counting neurons stained with a monoclonal antibody recognizing non-phosphorylated epitopes on neurofilaments (SMI-32) indicated that their number in the contralesional RNm was consistently inferior to that in the ipsilesional RNm, in a proportion amounting up to 35%. The lesion also induced shrinkage of the soma of the neurons detected in the contralesional RNm. Infusing an anti-Nogo-A antibody at the site of the lesion did not increase the proportion of SMI-32 positive rubrospinal neurons in the contralesional RNm nor prevent shrinkage.

A unilateral section of the corticospinal tract at cervical level in primate does not lead to measurable cell loss in motor cortex.

The effects of a unilateral interruption of the dorsolateral funiculus at cervical level on the survival of neurons in the motor cortex were investigated in macaque monkeys. The lesion was made on the left side at the transition region between the 7(th) and 8(th) cervical segments, above the motoneurons controlling hand muscles. As a result, the homolateral hand became paretic, although an incomplete recovery of manual dexterity took place during 2 months post-lesion. A quantitative anatomical assessment of pyramidal neurons in layer V was performed in the hindlimb area of the primary motor cortex and in the supplementary motor area (SMA proper). The pyramidal neurons were visualized using the marker SMI-32 and thus included the subpopulation of corticospinal neurons. These quantitative data demonstrated that the vast majority of the axotomized corticospinal (CS) neurons did not degenerate. Rather, their somata shrank, compared to the opposite hemisphere or to intact monkeys. This conclusion is in contrast to some previous studies in monkeys that argued for a substantial degeneration of motor cortex neurons as a result of transection of the corticospinal tract; yet in agreement with others that concluded the survival of most CS neurons. The survival of the majority of CS axotomized neurons is also consistent with the observation of numerous CS axons 1 mm above the cervical hemisection.

Arm to leg coordination in humans during walking, creeping and swimming activities.

In walking humans, arm to leg coordination is a well established phenomenon. The origin of this coordination, however, remains a matter for debate. It could derive from the intrinsic organisation of the human CNS, but it could also consist of a movement induced epiphenomenon. In order to establish which of these alternatives applies, we recorded arm and leg movements as well as their muscle activities during walking, creeping on all fours and swimming. The relationship between arm and leg cycle frequency observed under these various conditions was then investigated. We found that during walking, creeping on all fours or swimming, arm and leg movements remain frequency locked with a fixed relationship of 1/1, 2/1, 3/1, 4/1 or 5/1. When movements of the legs are slowed by flippers, the frequency relationship may skip to a different value, but the coordination is preserved. Furthermore, minimising the mechanical interactions between the limbs does not abolish coordination. These findings demonstrate that the arm to leg coordination observed in the walking human is also present during other human locomotor activities. The characteristics of this coordination correspond to those of a system of two coupled oscillators like that underlying quadruped locomotion.

Slow dorsal-ventral rhythm generator in the lamprey spinal cord.

In the isolated lamprey spinal cord, a very slow rhythm (0.03-0.11 Hz), superimposed on fast N-methyl-D-aspartate (NMDA)-induced locomotor activity (0.26-2.98 Hz), could be induced by a blockade of GABA(A) or glycine receptors or by administration of (1 s, 3 s)-l-aminocyclopentane-1,3-dicarboxylic acid a metabotropic glutamate receptor agonist. Ventral root branches supplying dorsal and ventral myotomes were exposed bilaterally to study the motor pattern in detail. The slow rhythm was expressed in two main forms: 1) a dorsal-ventral reciprocal pattern was the most common (18 of 24 preparations), in which bilateral dorsal branches were synchronous and alternated with the ventral branches, in two additional cases a diagonal dorsal-ventral reciprocal pattern with alternation between the left (or right) dorsal and the right (or left) ventral branches was observed; 2) synchronous bursting in all branches was encountered in four cases. In contrast, the fast locomotor rhythm occurred always in a left-right reciprocal pattern. Thus when the slow rhythm appeared in a dorsal-ventral reciprocal pattern, fast rhythms would simultaneously display left-right alternation. A longitudinal midline section of the spinal cord during ongoing slow bursting abolished the reciprocal pattern between ipsilateral dorsal and ventral branches but a synchronous burst activity could still remain. The fast swimming rhythm did not recover after the midline section. These results suggest that in addition to the network generating the swimming rhythm in the lamprey spinal cord, there is also a network providing slow reciprocal alternation between dorsal and ventral parts of the myotome. During steering, a selective activation of dorsal and ventral myotomes is required and the neural network generating the slow rhythm may represent activity in the spinal machinery used for steering.

Differential effects of the reticulospinal system on locomotion in lamprey.

Specific effects of stimulating different parts of the reticulospinal (RS) system on the spinal locomotor pattern are described in lamprey. In the in vitro brain stem and spinal cord preparation, microstimulation in different areas of the reticular formation was performed by ejecting a small amount of -glutamate from a micropipette. These areas were distributed over the four reticular nuclei of the brain stem: the mesencephalic reticular nucleus (MRN) and the anterior, middle and posterior rhombencephalic reticular nuclei (ARRN, MRRN, and PRRN, respectively). To prevent synaptic spread of excitation within the brain stem, the synaptic transmission was blocked by using a low Ca2+, high Mn2+ physiological saline in the brain stem pool. "Fictive" locomotion was evoked by applying N-methyl--aspartate (NMDA) to the spinal cord. Rhythmical discharges of motoneurons were recorded bilaterally in the midbody area, from the ventral roots that had been subdivided in dorsal and ventral branches, supplying the dorsal and ventral part of the myotome, respectively. Two major effects of brain stem stimulation were elicited: a change in the frequency of the locomotory rhythm and an induction of asymmetry (left/right, dorsal/ventral) in the segmental motor output. Approximately 50% of the stimulated sites evoked a change in locomotor frequency. In the PRRN almost all effective sites evoked an increase in frequency (10-50%). In the other nuclei, increase and decrease (10-30%) were observed equally frequently. Most of the stimulated sites (50-80%) in any reticular nucleus evoked asymmetry in the segmental motor output. Distortion of the segmental output symmetry was classified into eight categories by comparing the intensity of locomotor bursts in the dorsal and ventral branches of the two ventral roots, ipsilateral and contralateral to the stimulated side. These categories differed in the direction of the body flexion, which would be evoked during normal swimming: ipsilateral (I), contralateral (C), dorsal (D), ventral (V), ipsilateral and dorsal (ID), ipsilateral and ventral (IV), contralateral and dorsal (CD), and contralateral and ventral (CV). The different categories were not equally represented in each nucleus and across the nuclei. The most pronounced categories for each nucleus were as follow. In MRN: I (33%); ARRN: C (44%); MRRN: rostral part, I (36%) and caudal part, CV (42%); and PRRN: rostral part, I (40%) and caudal part, IV (35%). Other categories were also present but less common in each nucleus. To examine if the effects of brain stem stimulation were uniform along the spinal cord, recordings were performed from distal parts of the cord. Stimulation of a given point in the brain stem produced similar pattern of effects in 59% of cases and different patterns in 41% of cases. The main conclusion of the present study is that the proportion of RS neurons with different influences on the spinal locomotor network differs significantly among different parts of the reticular formation of the lamprey. The specificity of RS influences may represent a basis for modifications of the segmental locomotor output necessary for the control of equilibrium and steering during locomotion.

Pattern generation by two coupled time-discrete neural networks with synaptic depression.

Numerous animal behaviors, such as locomotion in vertebrates, are produced by rhythmic contractions that alternate between two muscle groups. The neuronal networks generating such alternate rhythmic activity are generally thought to rely on pacemaker cells or well-designed circuits consisting of inhibitory and excitatory neurons. However, experiments in organotypic cultures of embryonic rat spinal cord have shown that neuronal networks with purely excitatory and random connections may oscillate due to their synaptic depression, even without pacemaker cells. In this theoretical study, we investigate what happens if two such networks are symmetrically coupled by a small number of excitatory connections. We discuss a time-discrete mean-field model describing the average activity and the average synaptic depression of the two networks. Depending on the parameter values of the depression, the oscillations will be in phase, antiphase, quasiperiodic, or phase trapped. We put forward the hypothesis that pattern generators may rely on activity-dependent tuning of synaptic depression.

Reticulospinal neurones provide monosynaptic glycinergic inhibition of spinal neurones in lamprey.

In lamprey, distinct groups of reticulospinal neurones utilize different neurotransmitters such as glutamate or serotonin. The present study demonstrates that a group of reticulospinal neurones inhibit their target neurones by an action on glycine receptors. Simultaneous intracellular recordings from a reticulospinal neurone and spinal target neurone shows that the former may evoke an IPSP in the latter. These IPSPs are elicited at a constant latency and amplitude, and follow high frequency stimulation (100 Hz). Furthermore, the IPSPs are maintained when the excitatory amino acid synaptic transmission is blocked, suggesting that the effects are not elicited via a powerful disynaptic pathway. These data taken together establish the monosynaptic nature of the pathway. IPSPs elicited from single reticulospinal neurones or from electrical stimulation of the reticular formation are suppressed by administration of strychnine, suggesting that glycine is the neurotransmitter of these inhibitory reticulospinal neurones.

Cerebellar and cerebral inputs to corticocortical and corticofugal neurons in areas 5 and 7 in the cat.

1. In the parietal cortex (Px, areas 5 and 7), the organization and characteristics of cerebellar and cerebral inputs and their effects on efferent neurons were investigated with the use of intracellular and extracellular recording techniques in the anesthetized cat. 2. Evoked field potential analysis revealed that two regions of the Px, the caudal bank of the ansate sulcus (Ans. S.) and the crown of the suprasylvian gyrus (Ssyl. G.), received converging input from the dentate and the interpositus nucleus. The cerebellar input to the caudal bank of the Ans. S. was relayed via the ventrolateral region of the ventroanterior-ventrolateral (VA-VL) complex of the thalamus, whereas the cerebellar input to the crown of the Ssyl. G. was relayed via the dorsomedial region of the VA-VL complex. 3. A total of 176 neurons was recorded intracellularly in the Px to examine inputs from the cerebellum. Of these, 72 neurons were corticocortical neurons projecting to the motor cortex (Mx), and 48 were corticofugal neurons to the pontine nucleus (PN). Intracellular staining with horseradish peroxidase revealed that the former corticocortical neurons were layer III pyramidal neurons and the latter corticofugal neurons were layer V pyramidal neurons. 4. Stimulation of the brachium conjunctivum (BC) produced di- or polysynaptic excitatory postsynaptic potentials (EPSPs) in corticocortical neurons projecting to the Mx and corticofugal neurons to the pontine nucleus in the Px. The characteristics of BC-evoked EPSPs were different between the bank of the Ans. S. and the crown of the Ssyl. G. In the bank of the Ans. S., the slope of the rising phase of the BC-evoked EPSPs was steeper, and their minimum latency was shorter by 0.8 ms than those in the crown of the Ssyl. G. These differences may reflect differences in the terminal distribution and conduction velocity of the thalamocortical fibers relaying cerebellar input to these two parietal areas. 5. Stimulation of the Mx produced mono- or disynaptic EPSPs in both corticocortical neurons projecting to the Mx and corticofugal neurons projecting to the pontine nucleus in the Px. For each neuron, effective sites for inducing EPSPs were distributed very widely and sometimes covered both areas 4 and 6. Extensive corticocortical projection from the Mx to the Px was confirmed by injection of an anterograde tracer into the Mx. 6. These data indicate that neurons in the Px receive inputs from both the cerebellum and the Mx and send outputs to the Mx and the cerebellum.(ABSTRACT TRUNCATED AT 400 WORDS)

Rostro-caudal distribution of reticulospinal projections from different brainstem nuclei in the lamprey.

The reticulospinal (RS) system in the lamprey is responsible for the control of locomotion, postural corrections and steering. To perform these functions, the RS system has to affect different muscular compartments along the body axis selectively. In this study, the possibility that RS neurones in different nuclei may project to different parts of the spinal cord, was investigated. The rostro-caudal extent of single RS axons was defined by stimulating them antidromically while recording from their cell body. All recorded mesencephalic RS neurones projected to the caudal tip of the spinal cord. Of the rhombencephalic RS neurones, 26% of the recorded neurones did not reach the caudalmost fourth of the spinal cord and this proportion varied between the anterior (18%), middle (17%) and posterior (36%) rhombencephalic reticular nuclei. For these RS axons, the level of termination covered the whole rostro-caudal extent of the spinal cord. No correlation was found between the length of an axon and its conduction velocity or between the length of an axon and the rostro-caudal position of its cell body in the nuclei.

Thalamocortical organization in the cerebello-thalamo-cortical system.

In the cat, the cerebellum projects via the ventroanterior-ventrolateral (VA-VL) complex of the thalamus to the motor and premotor cortices and also to the parietal association cortex. Cerebellar inputs to each of these regions have been characterized electrophysiologically by depth profiles of cortical potentials following stimulation of the brachium conjunctivum and of the VA-VL complex, and morphologically by the laminar distribution of thalamocortical (TC) terminations, in aggregate and at the single-axon level. One population of TC neurons terminated mainly in layer I and was associated with late surface negative potentials. A second population, with terminations in layers III and IV, was associated with early deep negative potentials. Terminations in layer III of the motor cortex formed multiple patches about 1-1.5 mm wide (mediolateral), which aligned to form 2-5 mm stripes extended rostrocaudally. This pattern correlates with the configuration of individual TC axons, which have two to six separate terminal patches distributed over 6 mm (rostrocaudal). The wide morphological divergence of single TC axons in the cortex may imply functionally multiple innervation of different efferent columns. Alternately, along with other inputs, it may permit a highly dynamic output selection from multiple representations, for example, of a variety of muscle groups in different combinations.

Two modes of cerebellar input to the parietal cortex in the cat.

The characteristics of cerebellar input to the parietal cortex through the ventroanterior-ventrolateral (VA-VL) complex of the thalamus were investigated in the adult cat by using combined electrophysiological and anatomical methods. Two distinct parietal regions were activated by stimulation of the cerebellar nuclei (CN). In the first region located in the depth of the bank of the ansate sulcus, stimulation of the CN induced early surface positive-deep negative potentials and late surface negative-deep positive potentials. In this cortical area, potentials of similar shape and time course were evoked at a shorter latency by stimulation of the ventrolateral part of the VA-VL complex where large negative field potentials were evoked by stimulation of the CN. After injection of the anterograde tracer Phaseolus vulgaris leucoagglutinin (PHA-L) in this part of the VA-VL complex, axon terminals of thalamocortical (TC) fibers were found in layers I, III and IV in the depth of the bank of the ansate sulcus and layers I and III in the motor cortex. In the second region located in the suprasylvian gyrus, late surface negative-deep positive potentials were evoked by stimulation of the CN and similar potentials were evoked at a shorter latency from the dorsomedial part of the VA-VL complex where large cerebellar-evoked potentials could be recorded. PHA-L injection in this thalamic region stained TC fibers and their terminals in layer I of the suprasylvian gyrus, and in layers I and III of the motor cortex. The laminar distribution of TC axon terminals in two different regions of the parietal cortex could account for the depth profiles of the cerebellar- and the thalamic-evoked potentials in each region. These results show that cerebellar information is conveyed to two separate areas in the parietal cortex by two different TC pathways.

Contrasting properties of monkey somatosensory and motor cortex neurons activated during the control of force in precision grip.

1. Single cell activity was investigated in the precentral motor (MI) and postcentral somatosensory (SI) cortex of the monkey to compare the neuronal activity related to the control of isometric force in the precision grip and to assess the participation of SI in motor control. 2. Three monkeys (Macaca fascicularis) were trained in a visual step-tracking paradigm to generate and precisely maintain force on a transducer held between thumb and index finger. Great care was taken to have the monkeys use only their fingers without moving the wrist or proximal joints. In two monkeys electromyographic (EMG) activity was checked in 23 muscles over several sessions. 3. Five similar classes of task-related firing patterns were found in both SI and MI cortical hand and finger representations, but their relative proportions differed. The majority of the SI neurons were phasically or phasic-tonically active (61%), whereas in MI the neurons that decreased their firing rate with force were most frequent (42%). 4. The timing of activity changes related to the onset of force increase from low to higher levels strongly differed in the two neuronal populations. In SI, only 14% of the task-related neurons increased or decreased their firing rate before the onset of force increase, in contrast to 56% in MI. Only 3% of the SI neurons showed changes before the earliest EMG activation. 5. In both SI and MI neurons with tonic and phasic-tonic, increasing or decreasing discharge patterns disclosed a relationship between neuronal activity and static force. Distinction was made between neurons modulating their activity in a monotonic way and those that were active only at one force level and had a kind of recruitment or deactivation threshold. The latter ones were more frequent in MI than in SI, and in the neuron population with decreasing firing patterns. For the neurons with increases in activity, statistically significant linear correlations between firing rate and force were found more frequently in MI than in SI, where the proportion of nonsignificant correlations was relatively high (35% vs. 15% in MI). In SI the indexes of force sensitivity, calculated from the slopes of the regression lines, covered a wider range than in MI; and their distribution was bimodal, with one mean of 30 Hz/N and the other of 155 Hz/N. In contrast, the mean rate-force slope in MI was 69 Hz/N.(ABSTRACT TRUNCATED AT 400 WORDS)

Responses of motor cortex neurons to visual stimulation in the alert monkey.

Evidence for visually evoked activity in single neurons of area 4 is presented. These neurons modulated their activity in relation to stimuli moving across the visual field of the monkey, without showing habituation. Some of them even had direction sensitivity. Their activity was independent of eye movements and unrelated to muscle contractions. The visually activated neurons were distributed throughout the depth of motor cortex.

Sensorimotor cortical control of isometric force in the monkey.

Recordings from single neurones in the primary somatosensory (SI) and motor (MI) cortex of monkeys trained to precisely regulate force between thumb and index finger have disclosed the following contrasting properties between neurones in these two cortical regions: (1) the existence of neurones with similar discharge patterns within MI and SI but, between these regions, significantly different distributions of the classes of discharge patterns; (2) a late onset of activity change in SI neurones in relation to force increase as compared to significantly earlier changes in MI neurones; (3) linear relations between firing rate and isometric force for SI and MI neurones, however with a larger range of rate-force slopes in SI as compared to MI; (4) infrequent motor reactions to intracortical microstimulation in SI but frequent reactions in MI; (5) a majority of SI neurones with cutaneous afferent input in contrast to predominant input from deep tissues to MI neurones; and (6) context independent visually evoked activity observed exclusively in MI neurones. These major differences suggest that SI neuronal activity most likely reflects the input from peripheral receptors rather than, as postulated for MI neurones, the participation in movement initiation and the control of muscular contractions.

Neuronal activity in the postcentral cortex related to force regulation during a precision grip task.

Recordings from single neurons in the primary somatosensory cortex of the monkey during force regulation between the fingers showed following characteristics: the existence of classes of discharge patterns similar to those in motor cortex, but with differences in their distribution, a late onset of activity changes in relation to force increase and a linear relation to force, but with shallow mean rate-force slope.