Motor primitives are determined in early development and are then robustly conserved into adulthood.

Motor patterns in legged vertebrates show modularity in both young and adult animals, comprising motor synergies or primitives. Are such spinal modules observed in young mammals conserved into adulthood or altered? Conceivably, early circuit modules alter radically through experience and descending pathways' activity. We analyze lumbar motor patterns of intact adult rats and the same rats after spinal transection and compare these with adult rats spinal transected 5 days postnatally, before most motor experience, using only rats that never developed hind limb weight bearing. We use independent component analysis (ICA) to extract synergies from electromyography (EMG). ICA information-based methods identify both weakly active and strongly active synergies. We compare all spatial synergies and their activation/drive strengths as proxies of spinal modules and their underlying circuits. Remarkably, we find that spatial primitives/synergies of adult injured and neonatal injured rats differed insignificantly, despite different developmental histories. However, intact rats possess some synergies that differ significantly, although modestly, in spatial structure. Rats injured as adults were more similar in modularity to rats that had neonatal spinal transection than to themselves before injury. We surmise that spinal circuit modules for spatial synergy patterns may be determined early, before postnatal day 5 (P5), and remain largely unaltered by subsequent development or weight-bearing experience. An alternative explanation but equally important is that, after complete spinal transection, both neonatal and mature adult spinal cords rapidly converge to common synergy sets. This fundamental or convergent synergy circuitry, fully determined by P5, is revealed after spinal cord transection.

Teaching Adult Rats Spinalized as Neonates to Walk Using Trunk Robotic Rehabilitation: Elements of Success, Failure, and Dependence.

UNLABELLED: Robot therapy promotes functional recovery after spinal cord injury (SCI) in animal and clinical studies. Trunk actions are important in adult rats spinalized as neonates (NTX rats) that walk autonomously. Quadrupedal robot rehabilitation was tested using an implanted orthosis at the pelvis. Trunk cortical reorganization follows such rehabilitation. Here, we test the functional outcomes of such training. Robot impedance control at the pelvis allowed hindlimb, trunk, and forelimb mechanical interactions. Rats gradually increased weight support. Rats showed significant improvement in hindlimb stepping ability, quadrupedal weight support, and all measures examined. Function in NTX rats both before and after training showed bimodal distributions, with "poor" and "high weight support" groupings. A total of 35% of rats initially classified as "poor" were able to increase their weight-supported step measures to a level considered "high weight support" after robot training, thus moving between weight support groups. Recovered function in these rats persisted on treadmill with the robot both actuated and nonactuated, but returned to pretraining levels if they were completely disconnected from the robot. Locomotor recovery in robot rehabilitation of NTX rats thus likely included context dependence and/or incorporation of models of robot mechanics that became essential parts of their learned strategy. Such learned dependence is likely a hurdle to autonomy to be overcome for many robot locomotor therapies. Notwithstanding these limitations, trunk-based quadrupedal robot rehabilitation helped the rats to visit mechanical states they would never have achieved alone, to learn novel coordinations, and to achieve major improvements in locomotor function. SIGNIFICANCE STATEMENT: Neonatal spinal transected rats without any weight support can be taught weight support as adults by using robot rehabilitation at trunk. No adult control rats with neonatal spinal transections spontaneously achieve similar changes. The robot rehabilitation system can be inactivated and the skills that were learned persist. Responding rats cannot be detached from the robot altogether, a dependence develops in the skill learned. From data and analysis here, the likelihood of such rats to respond to the robot therapy can also now be predicted. These results are all novel. Understanding trunk roles in voluntary and spinal reflex integration after spinal cord injury and in recovery of function are broadly significant for basic and clinical understanding of motor function.

Trunk robot rehabilitation training with active stepping reorganizes and enriches trunk motor cortex representations in spinal transected rats.

Trunk motor control is crucial for postural stability and propulsion after low thoracic spinal cord injury (SCI) in animals and humans. Robotic rehabilitation aimed at trunk shows promise in SCI animal models and patients. However, little is known about the effect of SCI and robot rehabilitation of trunk on cortical motor representations. We previously showed reorganization of trunk motor cortex after adult SCI. Non-stepping training also exacerbated some SCI-driven plastic changes. Here we examine effects of robot rehabilitation that promotes recovery of hindlimb weight support functions on trunk motor cortex representations. Adult rats spinal transected as neonates (NTX rats) at the T9/10 level significantly improve function with our robot rehabilitation paradigm, whereas treadmill-only trained do not. We used intracortical microstimulation to map motor cortex in two NTX groups: (1) treadmill trained (control group); and (2) robot-assisted treadmill trained (improved function group). We found significant robot rehabilitation-driven changes in motor cortex: (1) caudal trunk motor areas expanded; (2) trunk coactivation at cortex sites increased; (3) richness of trunk cortex motor representations, as examined by cumulative entropy and mutual information for different trunk representations, increased; (4) trunk motor representations in the cortex moved toward more normal topography; and (5) trunk and forelimb motor representations that SCI-driven plasticity and compensations had caused to overlap were segregated. We conclude that effective robot rehabilitation training induces significant reorganization of trunk motor cortex and partially reverses some plastic changes that may be adaptive in non-stepping paraplegia after SCI.

Covert actions of epidural stimulation on spinal locomotor circuits.

Spinal circuitry produces the rhythm and patterning of locomotion. However, both descending and sensory inputs are required to initiate and adapt locomotion to the environment. Spinal cord injury (SCI) disrupts descending controls of the spinal cord, producing paralysis. Epidural stimulation (ES) is a promising clinical therapy for motor control recovery and is capable of reactivating the lumbar spinal locomotor networks, yet little is known about the effects of ES on locomotor neurons. Previously, we found that both sensory afferent pathways and serotonin exert mixed excitatory and inhibitory actions on lumbar interneurons involved in the generation of the locomotor rhythm, identified by the transcription factor Shox2. However, after chronic complete SCI, sensory afferent inputs to Shox2 interneurons become almost exclusively excitatory and Shox2 interneurons are supersensitive to serotonin. Here, we investigated the effects of ES on these SCI-induced changes. Inhibitory input from sensory pathways to Shox2 interneurons was maintained and serotonin supersensitivity was not observed in SCI mice that received daily sub-motor threshold ES. Interestingly, the effects of ES were maintained for at least three weeks after the ES was discontinued. In contrast, the effects of ES were not observed in Shox2 interneurons from mice that received ES after the establishment of the SCI-induced changes. Our results demonstrate mechanistic actions of ES at the level of identified spinal locomotor circuit neurons and the effectiveness of early treatment with ES on preservation of spinal locomotor circuitry after SCI, suggesting possible therapeutic benefits prior to the onset of motor rehabilitation.

Adaptation to a cortex-controlled robot attached at the pelvis and engaged during locomotion in rats.

Brain-machine interfaces (BMIs) should ideally show robust adaptation of the BMI across different tasks and daily activities. Most BMIs have used overpracticed tasks. Little is known about BMIs in dynamic environments. How are mechanically body-coupled BMIs integrated into ongoing rhythmic dynamics, for example, in locomotion? To examine this, we designed a novel BMI using neural discharge in the hindlimb/trunk motor cortex in rats during locomotion to control a robot attached at the pelvis. We tested neural adaptation when rats experienced (1) control locomotion, (2) "simple elastic load" (a robot load on locomotion without any BMI neural control), and (3) "BMI with elastic load" (in which the robot loaded locomotion and a BMI neural control could counter this load). Rats significantly offset applied loads with the BMI while preserving more normal pelvic height compared with load alone. Adaptation occurred over ∼100-200 step cycles in a trial. Firing rates increased in both the loaded conditions compared with baseline. Mean phases of the discharge of cells in the step cycle shifted significantly between BMI and the simple load condition. Over time, more BMI cells became positively correlated with the external force and modulated more deeply, and the network correlations of neurons on a 100 ms timescale increased. Loading alone showed none of these effects. The BMI neural changes of rate and force correlations persisted or increased over repeated trials. Our results show that rats have the capacity to use motor adaptation and motor learning to fairly rapidly engage hindlimb/trunk-coupled BMIs in their locomotion.

Modular organization of locomotor networks in people with severe spinal cord injury.

Introduction: Previous studies support modular organization of locomotor circuitry contributing to the activation of muscles in a spatially and temporally organized manner during locomotion. Human spinal circuitry may reorganize after spinal cord injury; however, it is unclear if reorganization of spinal circuitry post-injury affects the modular organization. Here we characterize the modular synergy organization of locomotor muscle activity expressed during assisted stepping in subjects with complete and incomplete spinal cord injury (SCI) of varying chronicity, before any explicit training regimen. We also investigated whether the synergy characteristics changed in two subjects who achieved independent walking after training with spinal cord epidural stimulation. Methods: To capture synergy structures during stepping, individuals with SCI were stepped on a body-weight supported treadmill with manual facilitation, while electromyography (EMGs) were recorded from bilateral leg muscles. EMGs were analyzed using non-negative matrix factorization (NMF) and independent component analysis (ICA) to identify synergy patterns. Synergy patterns from the SCI subjects were compared across different clinical characteristics and to non-disabled subjects (NDs). Results: Results for both NMF and ICA indicated that the subjects with SCI were similar among themselves, but expressed a greater variability in the number of synergies for criterion variance capture compared to NDs, and weaker correlation to NDs. ICA yielded a greater number of muscle synergies than NMF. Further, the clinical characteristics of SCI subjects and chronicity did not predict any significant differences in the spatial synergy structures despite any neuroplastic changes. Further, post-training synergies did not become closer to ND synergies in two individuals. Discussion: These findings suggest fundamental differences between motor modules expressed in SCIs and NDs, as well as a striking level of spatial and temporal synergy stability in motor modules in the SCI population, absent the application of specific interventions.

A neural basis for motor primitives in the spinal cord.

Motor primitives and modularity may be important in biological movement control. However, their neural basis is not understood. To investigate this, we recorded 302 neurons, making multielectrode recordings in the spinal cord gray of spinalized frogs, at 400, 800, and 1200 mum depth, at the L2/L3 segment border. Simultaneous muscle activity recordings were used with independent components analysis to infer premotor drive patterns. Neurons were divided into groups based on motor pattern modulation and sensory responses, depth recorded, and behavior. The 187 motor pattern modulated neurons recorded comprised 14 cutaneous neurons and 28 proprioceptive neurons at 400 mum in the dorsal horn, 131 intermediate zone interneurons from approximately 800 microm depth without sensory responses, and 14 motoneuron-like neurons at approximately 1200 microm. We examined all such neurons during spinal behaviors. Mutual information measures showed that cutaneous neurons and intermediate zone neurons were related better to premotor drives than to individual muscle activity. In contrast, proprioceptive-related neurons and ventral horn neurons divided evenly. For 46 of the intermediate zone interneurons, we found significant postspike facilitation effects on muscle responses using spike-triggered averages representing short-latency postspike facilitations to multiple motor pools. Furthermore, these postspike facilitations matched significantly in both their patterns and strengths with the weighting parameters of individual primitives extracted statistically, although both were initially obtained without reference to one another. Our data show that sets of dedicated interneurons may organize individual spinal primitives. These may be a key to understanding motor development, motor learning, recovery after CNS injury, and evolution of motor behaviors.

A simple experimentally based model using proprioceptive regulation of motor primitives captures adjusted trajectory formation in spinal frogs.

Spinal circuits may organize trajectories using pattern generators and synergies. In frogs, prior work supports fixed-duration pulses of fixed composition synergies, forming primitives. In wiping behaviors, spinal frogs adjust their motor activity according to the starting limb position and generate fairly straight and accurate isochronous trajectories across the workspace. To test whether a compact description using primitives modulated by proprioceptive feedback could reproduce such trajectory formation, we built a biomechanical model based on physiological data. We recorded from hindlimb muscle spindles to evaluate possible proprioceptive input. As movement was initiated, early skeletofusimotor activity enhanced many muscle spindles firing rates. Before movement began, a rapid estimate of the limb position from simple combinations of spindle rates was possible. Three primitives were used in the model with muscle compositions based on those observed in frogs. Our simulations showed that simple gain and phase shifts of primitives based on published feedback mechanisms could generate accurate isochronous trajectories and motor patterns that matched those observed. Although on-line feedback effects were omitted from the model after movement onset, our primitive-based model reproduced the wiping behavior across a range of starting positions. Without modifications from proprioceptive feedback, the model behaviors missed the target in a manner similar to that in deafferented frogs. These data show how early proprioception might be used to make a simple estimate initial limb state and to implicitly plan a movement using observed spinal motor primitives. Simulations showed that choice of synergy composition played a role in this simplicity. To generate froglike trajectories, a hip flexor synergy without sartorius required motor patterns with more proprioceptive knee flexor control than did patterns built with a more natural synergy including sartorius. Such synergy choices and control strategies may simplify the circuitry required for reflex trajectory construction and adaptation.

Individual premotor drive pulses, not time-varying synergies, are the units of adjustment for limb trajectories constructed in spinal cord.

Complex actions may arise by combining simple motor primitives. Our studies support individual premotor drive pulses or bursts as execution primitives in spinal cord. Alternatively, the fundamental execution primitives at the segmental level could be time-varying synergies. To distinguish these hypotheses, we examined sensory feedback effects during targeted wiping organized in spinal cord. This behavior comprises three bursts. We tested (1) whether feedback altered the structure of individual premotor drive bursts or primitives, and (2) whether feedback differentially modulated different drive bursts or pulses in the three burst sequence. At least two of the three bursts would need to always be comodulated to support a time-varying synergy. We used selective muscle vibration to control spindle feedback from a single muscle (biceps/iliofibularis). The structures of premotor drive bursts were conserved. However, biceps vibration (1) scaled the amplitudes of two bursts coactivated during the initial phase of wiping independently of one another without altering their phase, and (2) independently phase regulated the third burst but preserved its amplitude. Thus, all three bursts were regulated separately. Durations were unaffected. The independent effects depended on (1) time of vibration during wiping, (2) frequency of vibration, and (3) limb configuration. Because each of the three bursts was independently modulated, these data strongly support execution using individual premotor bursts rather than time-varying synergies at the spinal level of motor organization. Our data show that both sensory feedback and central systems of the spinal cord act in concert to adjust the individual premotor bursts in support of the straight and unimodal wiping trajectory.

Precise Tubular Braid Structures of Ultrafine Microwires as Neural Probes: Significantly Reduced Chronic Immune Response and Greater Local Neural Survival in Rat Cortex.

Braided multi-electrode probes (BMEPs) for neural interfaces comprise ultrafine microwire bundles interwoven into tubular braids. BMEPs provide highly flexible probes and tethers, and an open lattice structure with up to 24 recording/stimulating channels in precise geometries, currently all within a [Formula: see text] diameter footprint. This paper compares the long-term tissue effects of BMEPs ( [Formula: see text] wires) versus single conventional 50- [Formula: see text] wires, by testing nearby chronic immune response and neural survival in rat cortex. Four different types of electrodes were implanted in cortex in each of eight rats: 1) BMEP with tether; 2) tethered 50- [Formula: see text] wire; 3) BMEP without a tether; and 4) untethered 50- [Formula: see text] wire. Quantitative immunohistological statistical comparisons after eight weeks using GFAP, ED1, and NeuN staining clearly showed that both BMEP implants had significantly less tissue immune response and more neuronal survival than either of the 50- [Formula: see text] wires ( ) in each of the eight rats. Data strongly indicate that BMEP tissue responses are superior, and that BMEP designs partly alleviate chronic tissue inflammatory responses and neural losses. The flexible body, tether and open braid lattice, and finer wire diameters of BMEP designs may all contribute to reducing the biological long-term response.

Highly Flexible Precisely Braided Multielectrode Probes and Combinatorics for Future Neuroprostheses.

The braided multielectrode probe (BMEP) is an ultrafine microwire bundle interwoven into a precise tubular braided structure, which is designed to be used as an invasive neural probe consisting of multiple microelectrodes for electrophysiological neural recording and stimulation. Significant advantages of BMEPs include highly flexible mechanical properties leading to decreased immune responses after chronic implantation in neural tissue and dense recording/stimulation sites (24 channels) within the 100-200 µm diameter. In addition, because BMEPs can be manufactured using various materials in any size and shape without length limitations, they could be expanded to applications in deep central nervous system (CNS) regions as well as peripheral nervous system (PNS) in larger animals and humans. Finally, the 3D topology of wires supports combinatoric rearrangements of wires within braids, and potential neural yield increases. With the newly developed next generation micro braiding machine, we can manufacture more precise and complex microbraid structures. In this article, we describe the new machine and methods, and tests of simulated combinatoric separation methods. We propose various promising BMEP designs and the potential modifications to these designs to create probes suitable for various applications for future neuroprostheses.

Coordination strategies for limb forces during weight-bearing locomotion in normal rats, and in rats spinalized as neonates.

Some rats spinally transected as neonates (ST rats) achieve weight-supporting independent locomotion. The mechanisms of coordinated hind-limb weight support in such rats are not well understood. To examine these we compared ST rats (with better than 60% of weight supported steps) and normal rats that were trained to cross an instrumented runway. Ground reaction forces, coordination of hind-limb and forelimb forces and the motions of the center of pressure (CoP) were assessed. Normal rats crossed the runway with a diagonal trot. On average hind-limbs bore about 80% of the vertical load carried by forelimbs (45% body weight on hind-limbs 55% on forelimbs), although this varied. Forelimbs and hind-limbs acted synergistically to generate decelerative and propulsive rostrocaudal forces, which averaged 15% of body weight with maximums of 50%. Lateral forces were very small (<8% of body weight). Center of pressure progressed in jumps along a straight line with mean lateral deviations <1 cm. ST rats hind-limbs bore about 60% of the vertical load of forelimbs (37% body weight on hind-limbs, 63% on forelimbs), significantly less compared to intact rats (P < 0.05). ST rats showed similar mean rostrocaudal forces, but with significantly larger maximum fluctuations of up to 80% of body weight (P < 0.05). Joint force-plate recordings showed forelimbs and hind-limb rostrocaudal forces in ST rats were opposing and significantly different from intact rats (P < 0.05). Lateral forces were approximately 20% of body weight and significantly larger than in normal rats (P < 0.05). Center of pressure zig-zagged, with mean lateral deviations of approximately 2 cm and a significantly larger range (P < 0.05). The haunches were also observed to roll more than normal rats. The locomotor strategy of injured rats using limbs in opposition was presumably less efficient but their complex gait was statically stable. Because forelimbs and hind-limbs acted in opposition, the trunk was held compressed. Force coordination was likely managed largely by the voluntary control in forelimbs and trunk.

Modular premotor drives and unit bursts as primitives for frog motor behaviors.

Spinal cord modularity impacts on our understanding of reflexes, development, descending systems in normal motor control, and recovery from injury. We used independent component analysis and best-basis or matching pursuit wavepacket analysis to extract the composition and temporal structure of bursts in hindlimb muscles of frogs. These techniques make minimal a priori assumptions about drive and motor pattern structure. We compared premotor drive and burst structures in spinal frogs with less reduced frogs with a fuller repertoire of locomotory, kicking, and scratching behaviors. Six multimuscle drives explain most of the variance of motor patterns (approximately 80%). Each extracted drive was activated with pulses at a single time scale or common duration (approximately 275 msec) burst structure. The data show that complex behaviors in brainstem frogs arise as a result of focusing drives to smaller core groups of muscles. Brainstem drives were subsets of the muscle groups from spinal frogs. The 275 msec burst duration was preserved across all behaviors and was most precise in brainstem frogs. These data support a modular decomposition of frog behaviors into a small collection of unit burst generators and associated muscle drives in spinal cord. Our data also show that the modular organization of drives seen in isolated spinal cord is fine-tuned by descending controls to enable a fuller movement repertoire. The unit burst generators and their associated muscle synergies extracted here link the biomechanical "primitives," described earlier in the frog, rat, and cat, and to the elements of pattern generation examined in fictive preparations.

Decerebrate mammalian preparations: unalleviated or fully alleviated pain? A review and opinion.

In experimental decerebration of mammals, the cerebral cortex and thalamus are surgically or otherwise inactivated under traditional (pharmacologic) general anesthesia. Once the effects of the pharmacologic anesthesia have dissipated, the animal remains alive, but there is neither pain sensation nor consciousness. Because the Animal Welfare Act and its regulations recognize drugs as the only means to alleviate pain, it is unclear whether a decerebrate animal should be placed in U.S. Department of Agriculture (USDA) pain and distress category D (pain or distress alleviated by drugs) or E (unalleviated pain or distress). We present a rationale for including decerebrate animals in USDA category D. We also provide a general review of decerebration and suggestions for institutional animal care and use committees having to evaluate decerebration protocols.

Spinal primitives and intra-spinal micro-stimulation (ISMS) based prostheses: a neurobiological perspective on the "known unknowns" in ISMS and future prospects.

The current literature on Intra-Spinal Micro-Stimulation (ISMS) for motor prostheses is reviewed in light of neurobiological data on spinal organization, and a neurobiological perspective on output motor modularity, ISMS maps, stimulation combination effects, and stability. By comparing published data in these areas, the review identifies several gaps in current knowledge that are crucial to the development of effective intraspinal neuroprostheses. Gaps can be categorized into a lack of systematic and reproducible details of: (a) Topography and threshold for ISMS across the segmental motor system, the topography of autonomic recruitment by ISMS, and the coupling relations between these two types of outputs in practice. (b) Compositional rules for ISMS motor responses tested across the full range of the target spinal topographies. (c) Rules for ISMS effects' dependence on spinal cord state and neural dynamics during naturally elicited or ISMS triggered behaviors. (d) Plasticity of the compositional rules for ISMS motor responses, and understanding plasticity of ISMS topography in different spinal cord lesion states, disease states, and following rehabilitation. All these knowledge gaps to a greater or lesser extent require novel electrode technology in order to allow high density chronic recording and stimulation. The current lack of this technology may explain why these prominent gaps in the ISMS literature currently exist. It is also argued that given the "known unknowns" in the current ISMS literature, it may be prudent to adopt and develop control schemes that can manage the current results with simple superposition and winner-take-all interactions, but can also incorporate the possible plastic and stochastic dynamic interactions that may emerge in fuller analyses over longer terms, and which have already been noted in some simpler model systems.

Motor primitives--new data and future questions.

Motor primitives allow integration across scales in the motor system and may link movement construction and circuit organization. This review examines support for primitives, and new data relating primitives to concrete circuit elements across species. Both kinematic motor primitives and muscle synergy/kinetic motor primitives are reviewed. Motor primitives allow a modular hierarchy that may be re-used by volitional systems in novel ways. They can provide a developmental bootstrap for ethologically important actions. Collections of primitives somewhat constrain motor acts, but at the same time sets of primitives facilitate the rapid construction of these constrained actions, and can allow use of simpler controls. Novel motor skill likely requires augmentation to transcend the constraints present in initial collections of low level motor primitives. The benefits and limitations of motor primitives and the recognized knowledge gaps and needs for future research are briefly discussed.

Adaptation to elastic loads and BMI robot controls during rat locomotion examined with point-process GLMs.

Currently little is known about how a mechanically coupled BMI system's actions are integrated into ongoing body dynamics. We tested a locomotor task augmented with a BMI system driving a robot mechanically interacting with a rat under three conditions: control locomotion (BL), "simple elastic load" (E) and "BMI with elastic load" (BMI/E). The effect of the BMI was to allow compensation of the elastic load as a function of the neural drive. Neurons recorded here were close to one another in cortex, all within a 200 micron diameter horizontal distance of one another. The interactions of these close assemblies of neurons may differ from those among neurons at longer distances in BMI tasks and thus are important to explore. A point process generalized linear model (GLM), was used to examine connectivity at two different binning timescales (1 ms vs. 10 ms). We used GLM models to fit non-Poisson neural dynamics solely using other neurons' prior neural activity as covariates. Models at different timescales were compared based on Kolmogorov-Smirnov (KS) goodness-of-fit and parsimony. About 15% of cells with non-Poisson firing were well fitted with the neuron-to-neuron models alone. More such cells were fitted at the 1 ms binning than 10 ms. Positive connection parameters ("excitation" ~70%) exceeded negative parameters ("inhibition" ~30%). Significant connectivity changes in the GLM determined networks of well-fitted neurons occurred between the conditions. However, a common core of connections comprising at least ~15% of connections persisted between any two of the three conditions. Significantly almost twice as many connections were in common between the two load conditions (~27%), compared to between either load condition and the baseline. This local point process GLM identified neural correlation structure and the changes seen across task conditions in the rats in this neural subset may be intrinsic to cortex or due to feedback and input reorganization in adaptation.