Distinct temporal activity patterns in the rat M1 and red nucleus during skilled versus unskilled limb movement.

The mammalian motor system contains multiple interconnected supraspinal networks, but little is known about their relative roles in producing different movements and behaviors, particularly given their apparently fused activity in primates. We tested whether the task context, as well as using a phylogenetically older mammal, rats, could distinguish the separate contributions of these networks. We obtained simultaneous multi-single neuron recordings from the forelimb motor cortex and magnocellular red nucleus as rats performed two contextually different, but kinematically similar, forelimb reach-like tasks: highly learned, skilled reaching for food through a narrow slot, a task requiring extensive training, versus the swing phases of treadmill locomotion. In both the M1 and the mRN, large subpopulations of neurons peaked in their spike firing rates near the onset and the end of the swing phase during treadmill locomotion. In contrast, neural subgroups in the two areas displayed different temporal sequences of activity during the skilled reaching task. In the mRN, the majority of task-modulated neurons peaked in their firing rate in the middle of the reach when the rat was preparing to project the arm through the slot, whereas large subgroups of M1 neurons displayed elevated firing rates during the initial and terminal phases of the reach. These results suggest that motor-behavioral context can alter the degree of overlapping activity in different supraspinal sensorimotor networks. Moreover, results for the skilled reaching task in rats may have highlighted a distinct processing role of the rubral complex: adapting natural muscle synergies across joints and limbs to novel task demands, in concert with cortically based learning.

Real-time prediction of hand trajectory by ensembles of cortical neurons in primates.

Signals derived from the rat motor cortex can be used for controlling one-dimensional movements of a robot arm. It remains unknown, however, whether real-time processing of cortical signals can be employed to reproduce, in a robotic device, the kind of complex arm movements used by primates to reach objects in space. Here we recorded the simultaneous activity of large populations of neurons, distributed in the premotor, primary motor and posterior parietal cortical areas, as non-human primates performed two distinct motor tasks. Accurate real-time predictions of one- and three-dimensional arm movement trajectories were obtained by applying both linear and nonlinear algorithms to cortical neuronal ensemble activity recorded from each animal. In addition, cortically derived signals were successfully used for real-time control of robotic devices, both locally and through the Internet. These results suggest that long-term control of complex prosthetic robot arm movements can be achieved by simple real-time transformations of neuronal population signals derived from multiple cortical areas in primates.

Neural prosthetic devices for quadriplegia.

Research into the use of neural prosthetic devices to treat paralysing disorders is expanding rapidly. Functional electrical stimulation of muscles is now used to activate electrically paralysed muscles to restore movements such as the hand-grasp. Sensory feedback signals recorded from peripheral nerves are used to improve the control of functional electrical stimulation systems. Functional electrical stimulation may also be used to stimulate neural circuits in the spinal cord. Electrical recordings from the brain will soon improve patients' control of neural prostheses. Non-invasive electroencephalographic recordings can currently be used to move a cursor across a computer screen. The restoration of near-to-normal limb movement control will, however, require recordings from large numbers of neurons within the brain. Preliminary feasibility has been demonstrated in experimental animals.

Real-time control of a robot arm using simultaneously recorded neurons in the motor cortex.

To determine whether simultaneously recorded motor cortex neurons can be used for real-time device control, rats were trained to position a robot arm to obtain water by pressing a lever. Mathematical transformations, including neural networks, converted multineuron signals into 'neuronal population functions' that accurately predicted lever trajectory. Next, these functions were electronically converted into real-time signals for robot arm control. After switching to this 'neurorobotic' mode, 4 of 6 animals (those with > 25 task-related neurons) routinely used these brain-derived signals to position the robot arm and obtain water. With continued training in neurorobotic mode, the animals' lever movement diminished or stopped. These results suggest a possible means for movement restoration in paralysis patients.

Principal component analysis of neuronal ensemble activity reveals multidimensional somatosensory representations.

Principal components analysis (PCA) was used to define the linearly dependent factors underlying sensory information processing in the vibrissal sensory area of the ventral posterior medial (VPM) thalamus in eight awake rats. Ensembles of up to 23 single neurons were simultaneously recorded in this area, either during long periods of spontaneous behavior (including exploratory whisking) or controlled deflection of single whiskers. PCA rotated the matrices of correlation between these n neurons into a series of n uncorrelated principal components (PCs), each successive PC oriented to explain a maximum of the remaining variance. The fact that this transformation is mathematically equivalent to the general Hebb algorithm in linear neural networks provided a major rationale for performing it here on data from real neuronal ensembles. Typically, most information correlated across neurons in the ensemble was concentrated within the first 3-8 PCs. Each of these was found to encode distinct, and highly significant informational factors. These factor encodings were assessed in two ways, each making use of fact that each PC consisted of a matrix of weightings, one for each neuron. First, the neurons were rank ordered according to the locations of the central whiskers in their receptive fields, allowing their weightings within different PCs to be viewed as a function of their position within the whisker representation in the VPM. Each PC was found to define a distinctly different topographic mapping of the cutaneous surface. Next, the PCs were used to weight-sum the neurons' simultaneous activities to create population vectors (PVs). Each PV consisted of a single continuous time series which represented the expression of each PC's 'magnitude' in response to stimulation of different whiskers, or during behavioral events such as active tactile whisking. These showed that each PC functioned as a feature detector capable of selectively predicting significant sensory or behavioral events with far greater statistical reliability than could any single neuron. The encoding characteristics of the first few PCs were remarkably consistent across all animals and experimental conditions, including both spontaneous exploration and direct sensory stimulation: PC1 positively weighted all neurons, mainly according to their covariance. Thus it encoded global magnitude of ensemble activity, caused either by combined sensory inputs or intrinsic network activity, such as spontaneous oscillations. PC2 encoded spatial position contrast, generally in the rostrocaudal dimension, across the whole cutaneous surface represented by the ensemble. PC3 more selectively encoded contrast in an orthogonal (usually dorsoventral) dimension. A variable number of higher numbered PCs encoded local position contrast within one or more smaller regions of the cutaneous surface. The remaining PCs typically explained residual 'noise', i.e. the uncorrelated variance that constituted a major part of each neuron's activity. Differences in behavioral or sensory experience produced relatively little in the PC weighting patterns but often changed the variance they explained (eigenvalues) enough to alter their ordering. These results argue that PCA provides a powerful set of tools for selectively measuring neural ensemble activity within multiple functionally significant 'dimensions' of information processing. As such, it redefines the 'neuron' as an entity which contributes portions of its variance to processing not one, but several tasks.

Simultaneous encoding of tactile information by three primate cortical areas.

We used simultaneous multi-site neural ensemble recordings to investigate the representation of tactile information in three areas of the primate somatosensory cortex (areas 3b, SII and 2). Small neural ensembles (30-40 neurons) of broadly tuned somatosensory neurons were able to identify correctly the location of a single tactile stimulus on a single trial, almost simultaneously. Furthermore, each of these cortical areas could use different combinations of encoding strategies, such as mean firing rate (areas 3b and 2) or temporal patterns of ensemble firing (area SII), to represent the location of a tactile stimulus. Based on these results, we propose that ensembles of broadly tuned neurons, located in three distinct areas of the primate somatosensory cortex, obtain information about the location of a tactile stimulus almost concurrently.

Neonatal whisker removal reduces the discrimination of tactile stimuli by thalamic ensembles in adult rats.

Simultaneous recordings of up to 48 single neurons per animal were used to characterize the long-term functional effects of sensory plastic modifications in the ventral posterior medial nucleus (VPM) of the thalamus following unilateral removal of facial whiskers in newborn rats. One year after this neonatal whisker deprivation, neurons in the contralateral VPM responded to cutaneous stimulation of the face at much longer minimal latencies (15.2 +/- 8.2 ms, mean +/- SD) than did normal cells (8.8 +/- 5.3 ms) in the same subregion of the VPM. In 69% of these neurons, the initial sensory responses to stimulus offset were followed for up to 700 ms by reverberant trains of bursting discharge, alternating in 100-ms cycles with inhibition. Receptive fields in the deafferented VPM were also atypical in that they extended over the entire face, shoulder, forepaw, hindpaw, and even ipsilateral whiskers. Discriminant analysis (DA) was then used to statistically evaluate how this abnormal receptive field organization might affect the ability of thalamocortical neuronal populations to "discriminate" somatosensory stimulus location. To standardize this analysis, three stimulus targets ("groups") were chosen in all animals such that they triangulated the central region of the "receptive field" of the recorded multineuronal ensemble. In the normal animals these stimulus targets were whiskers or perioral hairs; in the deprived animals the targets typically included hairy skin of the body as well as face. The measured variables consisted of each neuron's spiking response to each stimulus differentiated into three poststimulus response epochs (0-15, 15-30, and 30-45 ms). DA quantified the statistical contribution of each of these variables to its overall discrimination between the three stimulus sites. In the normal animals, the stimulus locations were correctly classified in 88.2 +/- 3.7% of trials on the basis of the spatiotemporal patterns of ensemble activity derived from up to 18 single neurons. In the deprived animals, the stimulus locations were much less consistently discriminated (reduced to 73.5 +/- 12.6%; difference from controls significant at P < 0.01) despite the fact that much more widely spaced stimulus targets were used and even when up to 20 neurons were included in the ensemble. Overall, these results suggest that neonatal damage to peripheral sense organs may produce marked changes in the physiology of individual neurons in the somatosensory thalamus. Moreover, the present demonstration that these changes can profoundly alter sensory discrimination at the level of neural populations in the thalamus provides important evidence that the well-known perceptual effects of chronic peripheral deprivation may be partially attributable to plastic reorganization at subcortical levels.

Topographic and laminar organizations of the incertocortical pathway in rats.

The topographic and laminar organizations of the projection system from the zona incerta to the neocortex were studied by using both retrograde and anterograde methods in the rat. Injections of retrograde fluorescent tracers into different cortical areas revealed that the incertocortical projection neurons have a rough topographic organization with respect to their cortical targets. Furthermore, the incertocortical projecting neurons were found mainly in the dorsal and rostral subdivisions of the zona incerta, and none were found in the ventral subdivision. In cases which included three different fluorescent tracers injected into the frontal, the parietal and the occipital cortices, retrogradely single-labelled cells were found intermingled within the dorsal zona incerta. Very few double-labelled cells were noted, and triple-labelled cells were absent. Injections of anterograde tracers into the dorsal zona incerta demonstrate that labelled fibres traverse the striatum and terminate most densely in the outer half of layer I of the neocortex. The density of incertocortical terminals was greatest in the somatosensory cortex, while the innervation of visual cortical areas was sparse. Very fine and sparse bouton-like swellings of labelled incertocortical fibres were found running parallel along the pial surface. Since it has recently been shown that the incertocortical projections derive from GABAergic neurons, the present results suggest that the diffuse and roughly topographic projection from the zona incerta to the cerebral cortex may play an inhibitory role in widespread areas of cerebral cortex. This inhibitory action may preferentially target the distal dendrites of cortical neurons, since the majority of incertocortical terminals were found in the outer part of layer I of the neocortex.

Low-dose amphetamine elevates movement-related firing of rat striatal neurons.

To study the striatal role in amphetamine's stimulant effects on motor behavior, single neurons were recorded in the dorsolateral striatum of unrestrained rats before and after amphetamine injection (0.5 or 1.0 mg/kg, i.p.). Comparisons of firing were made between similar motor behaviors before and after injection. Mean locomotor firing rates increased 5% to 276% within 30 min after injection and reversed within 2 h. Firing related to specific head- or forelimb-movements, which were similar in all measured parameters before and after injection, was elevated several hundred percent after injection and then reversed, the time course paralleling that of the stimulant effect on these movements. Elevation of movement-related striatal firing rates by low doses of the psychomotor stimulant is in line with established increases in firing rate normally observed for striatal neurons related to motor behavior.

The estrous cycle and the olivo-cerebellar circuit. I. Contrast enhancement of sensorimotor-correlated cerebellar discharge.

Neuromodulation of Purkinje (Pnj) cell responses by monoamines and estrous hormones is well characterized in the cerebellum at the cellular level, but not at the level of neuronal circuits in the awake behaving animal. In the present study, simultaneous recordings of up to 16 single neurons from within the olivo-cerebellar circuit were obtained through chronically implanted microwire electrode bundles: arrays of Pnj cell like neurons (Pnj cln) in the paravermal cerebellum and neurons within the afferent source of its climbing fiber input, the rostral dorsal accessory olive (rDAO), were recorded simultaneously across 3-20 consecutive estrous cycles during constant or variable speed treadmill locomotion performance tasks. Over 90% of Pnj cln recorded during treadmill locomotion exhibited significant increases (80%) or decreases (10%) in activity correlated with the stance phase of locomotion. In contrast, cells from the rDAO increased activity during speed changes or when the rat failed to maintain the treadmill speed (position slip). On the night of behavioral estrus, which is triggered by elevations in circulating levels of 17 beta-estradiol and progesterone, the magnitude of both increases and decreases in stance-correlated Pnj cln activity increased by 85-115%. These results are consistent with our previous findings that 17 beta-estradiol and progesterone enhance excitatory and inhibitory responses of single Pnj cells to locally applied glutamate and GABA, respectively. This dual enhancement of both excitatory and inhibitory effects, apparently paradoxical at the cellular level, produced a marked heightening of the contrast of the neural population "signal" at the neuronal ensemble level. Furthermore, the stance-correlated discharge of Pnj cln during estrus preceded that during diestrus by approximately 120 ms. Frame-by-frame video analysis also suggested that the swing phase of the step cycle was shortened on estrus compared with diestrus (low hormone state). In addition, rDAO discharge correlated with speed change or position slip was also significantly increased (P < 0.05) on the night of behavioral estrus versus diestrus. Thus, estrus was associated with changes in both the amplitude and the timing of Pnj cln and rDAO discharge correlated with specific behavioral events. These estrous-associated changes in Pnj cell activity were well correlated (r = 0.84) with faster responses to random changes in treadmill speed, a motor performance task. Together, these findings suggest that the increases in the contrast of stance-correlated Phj cln discharge observed following peak circulating levels of sex steroid hormones are associated with improved motor performance on a randomly moving treadmill.

The estrous cycle and the olivo-cerebellar circuit. II. Enhanced selective sensory gating of responses from the rostral dorsal accessory olive.

This study demonstrates that gating of responses of the rostral dorsal accessory olive (rDAO) to somatosensory stimulation varies across the estrous hormone cycle of the rat. The rDAO has been suggested as an "error" or event signal generator for the cerebellar cortex. Selective sensory gating of input to this structure may underlie this error signalling function. In the present study, as many as 23 single neurons were recorded simultaneously from either the forepaw or the snout areas of the rDAO. Responses of these neurons to electrical stimulation of peripheral afferents were determined during active movement or non-movement conditions. These results were then compared across the estrous cycle or after administration of the estrous hormones 17 beta-estradiol (E2) and/or progesterone (P) to rats on diestrus or following E2 priming. Elevations in circulating estrous hormones produced greater excitatory responses of rDAO neurons to stimulation during non-movement, and, conversely, enhanced inhibition of rDAO activity during active movement of the stimulated peripheral area compared with control diestrous conditions, suggesting that selective gating processes to the rDAO are enhanced by estrous hormones. The results of this study suggest that the night of behavioral estrus is associated with enhanced selective sensory gating processes associated with improved detection and processing of error signals.

Active tactile exploration influences the functional maturation of the somatosensory system.

1. The hypothesis that active exploration of objects is required for the functional maturation of neuronal circuits subserving tactile perception was tested by subjecting 8- to 11-day old rats to a complete unilateral section of the facial nerve. This procedure selectively abolished whisker protraction movements without affecting the sensory innervation of the facial vibrissae, the tactile organs used by rats to discriminate object texture and shape. 2. Six to 14 mo after the facial nerve section, simultaneous recordings of neuronal ensembles located in the ventral posterior medial nucleus (VPM) of the thalamus revealed a marked reduction in receptive field (RF) size (in terms of number of whiskers), and the formation of abnormal RF surrounds, spanning the face and contiguous body regions. In addition, the directional organization of VPM RFs, represented by caudal to rostral shifts in RF centers over 30 ms following whisker stimulation, was greatly reduced in these animals. 3. These results suggest that neonatal active tactile exploration is required to establish normal spatiotemporal patterning of neuronal RFs within the somatosensory system, and consequently, to develop normal tactile perception.

Calbindin-containing non-specific thalamocortical projecting neurons in the rat.

Immunoreactivity for calcium binding proteins was used to demonstrate the neurochemical profiles of non-specific thalamocortical neurons located in the ventromedial nucleus, the centrolateral nucleus, and the nucleus reuniens that project to the somatosensory cortex in the adult rat. Cortical injections of fluorescent tracers combined with immunohistochemistry for calcium binding proteins revealed that retrogradely labeled neurons in these three thalamic nuclei are immunoreactive for calbindin. The present results suggest the presence of a chemically distinct non-specific thalamocortical system which terminates in the neocortex.

Effects of systemic and local ethanol on responses of rat cerebellar Purkinje neurons to iontophoretically applied gamma-aminobutyric acid.

The goals of this study were: (1) to determine the effects of acute systemic or local application of ethanol (ETOH) on the response of cerebellar Purkinje cells (P-cells) to iontophoretically applied gamma-aminobutyric acid (GABA) and (2) to characterize the effects of Ro15-4513, a putative antagonist of ETOH-GABA interactions, on ETOH-induced changes in GABA responsiveness. Male Sprague-Dawley rats (230-370 g) were anesthetized with halothane and implanted with intraperitoneal catheters for administration of ETOH (1.0-2.0 g/kg), before the recording session. Extracellular activity of single P-cells was recorded with the central barrel of a five-barrel micropipette, the other barrels of which were used for microiontophoresis of GABA and electro-osmosis of ETOH at the recording site. Spontaneous discharge and response of P-cells to GABA were monitored during a pre-ETOH control and for 1-1.5 h after systemic or electro-osmotic administration of ETOH. Transient suppression of spontaneous P-cell discharge was usually observed within 4-8 min of systemic ETOH injection. This effect lasted 2-4 min in 10 out of 19 rats tested. GABA-mediated inhibitory responses of cerebellar P-cells were increased by 45-50% relative to pre-ETOH values at 10 and 90 min post-ETOH injection. Prior administration of the imidazobenzodiazepine Ro15-4513 (4-6 mg/kg) failed to antagonize either the ETOH-induced enhancement of GABA-mediated inhibition or the transient inhibition of spontaneous P-cell activity rat cerebellar P-cell produced by ETOH. In these studies, electro-osmotically applied ETOH produced a potent suppression of spontaneous P-cell activity which precluded further augmentation of unit responses to GABA. These results show that doses of systemically administered ETOH which are mildly intoxicating in the awake, behaving animal, enhance the inhibitory action of GABA on cerebellar P-cell discharge.

Effects of systemic and local ethanol on responses of rat cerebellar Purkinje neurons to iontophoretically applied norepinephrine and gamma-aminobutyric acid.

The goal of the present study was to determine the effect of acute ethanol (ETOH), administered intraperitoneally or electro-osmotically, on norepinephrine (NE) induced increases in gamma-aminobutyric acid (GABA) mediated inhibition of single cerebellar Purkinje neurons (P-cells). Male Sprague-Dawley rats (230-370g) were anesthetized with halothane and implanted with an intraperitoneal catheter for systemic administration of ETOH (1.0-1.5 g/kg) prior to the recording session. Extracellular activity of single P-cells was recorded before and after iontophoresis of GABA and NE using five-barrel glass micropipettes. GABA was administered at the recording site by microiontophoretic pulses before, during and after continuous iontophoretic application of NE. Spontaneous discharge, GABA responses and NE-GABA interactions in P-cells were monitored for each experiment before and 1-1.5 h following systemic administration of ETOH. As in our previous reports administration of NE, at low ejection currents (10-60 nA), augmented GABA mediated suppression of P-cell spontaneous discharge. Between 10 and 60 min after injection of ETOH, this NE induced augmentation of GABA inhibition was further potentiated. This potentiation involved increases in both the magnitude and the duration of the GABA inhibition observed after NE alone. NE-induced augmentation of GABA inhibition persisted for 2-13 min longer after ETOH administration than in the pre-ETOH control period. Local electro-osmotic application of ETOH, which resulted in strong depression of spontaneous activity and caused small increases in GABA-mediated inhibition, did not directly potentiate NE-induced augmentation of GABA action. These results indicate that NE-mediated augmentation of GABA inhibition of P-cell activity is potentiated following systemic, but not local, ETOH administration.

Sensorimotor encoding by synchronous neural ensemble activity at multiple levels of the somatosensory system.

Neural ensemble processing of sensorimotor information during behavior was investigated by simultaneously recording up to 48 single neurons at multiple relays of the rat trigeminal somatosensory system. Cortical, thalamic, and brainstem neurons exhibited widespread 7- to 12-hertz synchronous oscillations, which began during attentive immobility and reliably predicted the imminent onset of rhythmic whisker twitching. Each oscillatory cycle began as a traveling wave of neural activity in the cortex that then spread to the thalamus. Just before the onset of rhythmic whisker twitching, the oscillations spread to the spinal trigeminal brainstem complex. Thereafter, the oscillations at all levels were synchronous with whisker protraction. Neural structures manifesting these rhythms also exhibited distributed spatiotemporal patterns of neuronal ensemble activity in response to tactile stimulation. Thus, multilevel synchronous activity in this system may encode not only sensory information but also the onset and temporal domain of tactile exploratory movements.

Development of direct GABAergic projections from the zona incerta to the somatosensory cortex of the rat.

The postnatal development of direct thalamocortical projections from the zona incerta of the ventral thalamus to the whisker representation area of the rat primary somatosensory cortex was investigated. Cytoarchitectonic analysis based on Nissl staining, cytochrome oxidase histochemistry and immunohistochemistry for glutamic acid decarboxylase, GABA, parvalbumin and calbindin D28K revealed that the zona incerta can be clearly distinguished from surrounding diencephalic structures from the day of birth. Moreover, four distinct anatomical subdivisions of this nucleus were identified: the rostral, dorsal, ventral and caudal. Of these, the ventral subdivision is by far the most conspicuous, containing the highest density of neurons, and the highest levels of cytochrome oxidase, glutamate decarboxylase, GABA, parvalbumin and calbindin D28K. In contrast, the dorsal, rostral and caudal subdivisions contain fewer cells, lower levels of glutamic acid decarboxylase and GABA and very few parvalbumin-positive and calbindin-positive neurons. Small injections of rhodamine coated microspheres or Fluoro-gold in the primary somatosensory cortex of animals at different stages of development revealed the existence of retrogradely labeled neurons in the rostral and dorsal subdivisions of the zona incerta from postnatal day 1. At this age, retrogradely labeled cells were also found in the ventral lateral, ventral posterior medial, posterior medial, centrolateral, ventral medial and magnocellular subdivision of the medial geniculate nuclei of the dorsal thalamus. The density of the incertocortical projection reaches its maximum between the first and second postnatal weeks, decreasing subsequently, until an adult pattern of labeling is achieved. Tracer injections combined with immunohistochemistry revealed that the majority of the incertocortical projection derives from GABAergic neurons, implying a potentially inhibitory role for the incertocortical projection. These results demonstrate that the rat trigeminal system contains parallel thalamocortical pathways of opposite polarity, emerging from both the dorsal (glutamatergic, excitatory) and ventral (GABAergic, inhibitory) thalamus since the day of birth. As such, these findings suggest that, contrary to the classical notion, not only the dorsal but also the ventral thalamus may play a special role in both cortical maturation and function.