Roles of the cerebellar vermis in predictive postural controls against external disturbances.

The central nervous system predictively controls posture against external disturbances; however, the detailed mechanisms remain unclear. We tested the hypothesis that the cerebellar vermis plays a substantial role in acquiring predictive postural control by using a standing task with floor disturbances in rats. The intact, lesioned, and sham groups of rats sequentially underwent 70 conditioned floor-tilting trials, and kinematics were recorded. Six days before these recordings, only the lesion group underwent focal suction surgery targeting vermal lobules IV-VIII. In the naïve stage of the sequential trials, the upright postures and fluctuations due to the disturbance were mostly consistent among the groups. Although the pattern of decrease in postural fluctuation due to learning corresponded among the groups, the learning rate estimated from the lumbar displacement was significantly lower in the lesion group than in the intact and sham groups. These results suggest that the cerebellar vermis contributes to predictive postural controls.

Optical Myography-Based Sensing Methodology of Application of Random Loads to Muscles during Hand-Gripping Training.

Hand-gripping training is important for improving the fundamental functions of human physical activity. Bernstein's idea of "repetition without repetition" suggests that motor control function should be trained under changing states. The randomness level of load should be visualized for self-administered screening when repeating various training tasks under changing states. This study aims to develop a sensing methodology of random loads applied to both the agonist and antagonist skeletal muscles when performing physical tasks. We assumed that the time-variability and periodicity of the applied load appear in the time-series feature of muscle deformation data. In the experiment, 14 participants conducted the gripping tasks with a gripper, ball, balloon, Palm clenching, and paper. Crumpling pieces of paper (paper exercise) involves randomness because the resistance force of the paper changes depending on the shape and layers of the paper. Optical myography during gripping tasks was measured, and time-series features were analyzed. As a result, our system could detect the random movement of muscles during training.

Rats adaptively seek information to accommodate a lack of information.

Metacognition is the ability to adaptively control one's behavior by referring to one's own cognitive processes. It is thought to contribute to learning in situations where there is insufficient information available from the environment. Information-seeking behavior is a type of metacognition in which one confirms the necessary information only when one does not have the necessary and sufficient information to accomplish a task. The rats were required to respond to a nose poke hole on one wall of the experimental box for a certain period of time and then move to the opposite side at a specific time. Unfortunately, they were unable to match the timing when responding to the hole on one side. Therefore, they had to look back and confirm that now was the right time. The results obtained by analyzing these looking-back movements using a motion capture system showed that this behavior occurred frequently and rapidly in situations of insufficient information, such as in the early stages of learning, but was hardly observed and became slower as learning progressed. These results suggest that rats can adjust their behavior in response to a lack of information more flexibly than previously assumed.

Gait disorders induced by photothrombotic cerebellar stroke in mice.

Patients with cerebellar stroke display relatively mild ataxic gaits. These motor deficits often improve dramatically; however, the neural mechanisms of this improvement have yet to be elucidated. Previous studies in mouse models of gait ataxia, such as ho15J mice and cbln1-null mice, have shown that they have a dysfunction of parallel fiber-Purkinje cell synapses in the cerebellum. However, the effects of cerebellar stroke on the locomotor kinematics of wild-type mice are currently unknown. Here, we performed a kinematic analysis of gait ataxia caused by a photothrombotic stroke in the medial, vermal, and intermediate regions of the cerebellum of wild-type mice. We used the data and observations from this analysis to develop a model that will allow locomotive prognosis and indicate potential treatment regimens following a cerebellar stroke. Our analysis showed that mice performed poorly in a ladder rung test after a stroke. During walking on a treadmill, the mice with induced cerebellar stroke had an increased duty ratio of the hindlimb caused by shortened duration of the swing phase. Overall, our findings suggest that photothrombotic cerebellar infarction and kinematic gait analyses will provide a useful model for quantification of different types of acute management of cerebellar stroke in rodents.

Contribution of Afferent Feedback to Adaptive Hindlimb Walking in Cats: A Neuromusculoskeletal Modeling Study.

Mammalian locomotion is generated by central pattern generators (CPGs) in the spinal cord, which produce alternating flexor and extensor activities controlling the locomotor movements of each limb. Afferent feedback signals from the limbs are integrated by the CPGs to provide adaptive control of locomotion. Responses of CPG-generated neural activity to afferent feedback stimulation have been previously studied during fictive locomotion in immobilized cats. Yet, locomotion in awake, behaving animals involves dynamic interactions between central neuronal circuits, afferent feedback, musculoskeletal system, and environment. To study these complex interactions, we developed a model simulating interactions between a half-center CPG and the musculoskeletal system of a cat hindlimb. Then, we analyzed the role of afferent feedback in the locomotor adaptation from a dynamic viewpoint using the methods of dynamical systems theory and nullcline analysis. Our model reproduced limb movements during regular cat walking as well as adaptive changes of these movements when the foot steps into a hole. The model generates important insights into the mechanism for adaptive locomotion resulting from dynamic interactions between the CPG-based neural circuits, the musculoskeletal system, and the environment.

A Model of Predictive Postural Control Against Floor Tilting in Rats.

Humans and animals learn the internal model of bodies and environments from their experience and stabilize posture against disturbances based on the predicted future states according to the internal model. We evaluated the mechanism of predictive control during standing, by using rats to construct a novel experimental system and comparing their behaviors with a mathematical model. In the experiments, rats (n = 6) that were standing upright using their hindlimbs were given a sensory input of light, after a certain period, the floor under them tilted backward. Initially, this disturbance induced a large postural response, including backward rotation of the center-of-mass angle and hindlimb segments. However, the rats gradually adjusted to the disturbance after experiencing 70 sequential trials, and a reduction in the amplitude of postural response was noted. We simulated the postural control of the rats under disturbance using an inverted pendulum model and model predictive control (MPC). MPC is a control method for predicting the future state using an internal model of the control target. It provides control inputs that optimize the predicted future states. Identification of the predictive and physiological parameters so that the simulation corresponds to the experiment, resulted in a value of predictive horizon (0.96 s) close to the interval time in the experiment (0.9-1.15 s). These results suggest that the rats predict posture dynamics under disturbance based on the timing of the sensory input and that the central nervous system provides plasticity mechanisms to acquire the internal model for MPC.

Quantitative evaluation of posture control in rats with inferior olive lesions.

Impairment of inferior olivary neurons (IONs) affects whole-body movements and results in abnormal gait and posture. Because IONs are activated by unpredicted motion rather than regular body movements, the postural dysfunction caused by ION lesions is expected to involve factors other than simple loss of feedback control. In this study, we measured the postural movements of rats with pharmacological ION lesions (IO rats) trained to stand on their hindlimbs. The coordination of body segments as well as the distribution and frequency characteristics of center of mass (COM) motion were analyzed. We determined that the lesion altered the peak properties of the power spectrum density of the COM, whereas changes in coordination and COM distribution were minor. To investigate how the observed properties reflected changes in the control system, we constructed a mathematical model of the standing rats and quantitatively identified the control system. We found an increase in linear proportional control and a decrease in differential and nonlinear control in IO rats compared with intact rats. The dystonia-like changes in body stiffness explain the nature of the linear proportional and differential control, and a disorder in the internal model is one possible cause of the decrease in nonlinear control.

Phase-Dependent Response to Afferent Stimulation During Fictive Locomotion: A Computational Modeling Study.

Central pattern generators (CPGs) in the spinal cord generate rhythmic neural activity and control locomotion in vertebrates. These CPGs operate under the control of sensory feedback that affects the generated locomotor pattern and adapt it to the animal's biomechanics and environment. Studies of the effects of afferent stimulation on fictive locomotion in immobilized cats have shown that brief stimulation of peripheral nerves can reset the ongoing locomotor rhythm. Depending on the phase of stimulation and the stimulated nerve, the applied stimulation can either shorten or prolong the current locomotor phase and the locomotor cycle. Here, we used a mathematical model of a half-center CPG to investigate the phase-dependent effects of brief stimulation applied to CPG on the CPG-generated locomotor oscillations. The CPG in the model consisted of two half-centers mutually inhibiting each other. The rhythmic activity in each half-center was based on a slowly inactivating, persistent sodium current. Brief stimulation was applied to CPG half-centers in different phases of the locomotor cycle to produce phase-dependent changes in CPG activity. The model reproduced several results from experiments on the effect of afferent stimulation of fictive locomotion in cats. The mechanisms of locomotor rhythm resetting under different conditions were analyzed using dynamic systems theory methods.

Gait Generation and Its Energy Efficiency Based on Rat Neuromusculoskeletal Model.

Changing gait is crucial for adaptive and smooth animal locomotion. Although it remains unclear what makes animals decide on a specific gait, energy efficiency is an important factor. It has been reported that the relationship of oxygen consumption with speed is U-shaped for each horse gait and that different gaits have different speeds at which oxygen consumption is minimized. This allows the horse to produce energy-efficient locomotion in a wide speed range by changing gait. However, the underlying mechanisms causing oxygen consumption to be U-shaped and the speeds for the minimum consumption to be different between different gaits are unclear. In the present study, we used a neuromusculoskeletal model of the rat to examine the mechanism from a dynamic viewpoint. Specifically, we constructed the musculoskeletal part of the model based on empirical anatomical data on rats and the motor control model based on the physiological concepts of the spinal central pattern generator and muscle synergy. We also incorporated the posture and speed regulation models at the levels of the brainstem and cerebellum. Our model achieved walking through forward dynamic simulation, and the simulated joint kinematics and muscle activities were compared with animal data. Our model also achieved trotting by changing only the phase difference of the muscle-synergy-based motor commands between the forelimb and hindlimb. Furthermore, the speed of each gait varied by changing only the extension phase duration and amplitude of the muscle synergy-based motor commands and the reference values for the regulation models. The relationship between cost of transport (CoT) and speed was U-shaped for both the generated walking and trotting, and the speeds for the minimum CoT were different for the two gaits, as observed in the oxygen consumption of horses. We found that the resonance property and the posture and speed regulations contributed to the CoT shape and difference in speeds for the minimum CoT. We further discussed the energy efficiency of gait based on the simulation results.

Adaptive hindlimb split-belt treadmill walking in rats by controlling basic muscle activation patterns via phase resetting.

To investigate the adaptive locomotion mechanism in animals, a split-belt treadmill has been used, which has two parallel belts to produce left-right symmetric and asymmetric environments for walking. Spinal cats walking on the treadmill have suggested the contribution of the spinal cord and associated peripheral nervous system to the adaptive locomotion. Physiological studies have shown that phase resetting of locomotor commands involving a phase shift occurs depending on the types of sensory nerves and stimulation timing, and that muscle activation patterns during walking are represented by a linear combination of a few numbers of basic temporal patterns despite the complexity of the activation patterns. Our working hypothesis was that resetting the onset timings of basic temporal patterns based on the sensory information from the leg, especially extension of hip flexors, contributes to adaptive locomotion on the split-belt treadmill. Our hypothesis was examined by conducting forward dynamic simulations using a neuromusculoskeletal model of a rat walking on a split-belt treadmill with its hindlimbs and by comparing the simulated motions with the measured motions of rats.

Alteration of gait parameters in a mouse model of surgically induced knee osteoarthritis.

PURPOSE: Joint pain is the most common symptom of osteoarthritis (OA); however, its mechanism remains unclarified. The present study investigated hindlimb motion during locomotion on the treadmill using a three-dimensional (3D) motion analysis system with high-speed cameras to evaluate whether this method can be used as an indication of joint pain in a mouse model of surgically induced OA. METHODS: We resected the medial meniscus and medial collateral ligament in 8-week old C57BL/6 male mice and performed locomotion recording 6 months post-operatively. Additionally, we performed the same recording after oral administration of the selective cyclooxygenase-2 inhibitor to determine whether alteration of the parameters were associated with joint pain. RESULTS: OA development, characterized by cartilage degeneration and osteophyte formation, was markedly enhanced in the OA group. There was no significant difference between the sham and OA groups in basic gait parameters, including stance duration, swing duration and gait cycle. However, when we divided the gait cycle into four phases and calculated the joint ranges of motion in each phase, the range of motion of the knee joint during the stepping-in phase and the swing duration were significantly decreased in the OA group. These significant differences between the sham and OA groups were diminished by the oral administration of a selective cyclooxygenase-2 inhibitor to the OA group. CONCLUSION: The present method may be useful to evaluate joint pain in experimental mice and contribute to elucidating the molecular mechanisms of pain in the OA knee joint in combination with genetically modified mice.

Improvement of cerebellar ataxic gait by injecting Cbln1 into the cerebellum of cbln1-null mice.

Patients and rodents with cerebellar damage display ataxic gaits characterized by impaired coordination of limb movements. Here, gait ataxia in mice with a null mutation of the gene for the cerebellin 1 precursor protein (cbln1-null mice) was investigated by kinematic analysis of hindlimb movements during locomotion. The Cbln1 protein is predominately produced and secreted from cerebellar granule cells. The cerebellum of cbln1-null mice is characterized by an 80% reduction in the number of parallel fiber-Purkinje cell synapses compared with wild-type mice. Our analyses identified prominent differences in the temporal parameters of locomotion between cbln1-null and wild-type mice. The cbln1-null mice displayed abnormal hindlimb movements that were characterized by excessive toe elevation during the swing phase, and by severe hyperflexion of the ankles and knees. When recombinant Cbln1 protein was injected into the cerebellum of cbln1-null mice, the step cycle and stance phase durations increased toward those of wild-type mice, and the angular excursions of the knee during a cycle period showed a much closer agreement with those of wild-type mice. These findings suggest that dysfunction of the parallel fiber-Purkinje cell synapses might underlie the impairment of hindlimb movements during locomotion in cbln1-null mice.

Postural control during quiet bipedal standing in rats.

The control of bipedal posture in humans is subject to non-ideal conditions such as delayed sensation and heartbeat noise. However, the controller achieves a high level of functionality by utilizing body dynamics dexterously. In order to elucidate the neural mechanism responsible for postural control, the present study made use of an experimental setup involving rats because they have more accessible neural structures. The experimental design requires rats to stand bipedally in order to obtain a water reward placed in a water supplier above them. Their motions can be measured in detail using a motion capture system and a force plate. Rats have the ability to stand bipedally for long durations (over 200 s), allowing for the construction of an experimental environment in which the steady standing motion of rats could be measured. The characteristics of the measured motion were evaluated based on aspects of the rats' intersegmental coordination and power spectrum density (PSD). These characteristics were compared with those of the human bipedal posture. The intersegmental coordination of the standing rats included two components that were similar to that of standing humans: center of mass and trunk motion. The rats' PSD showed a peak at approximately 1.8 Hz and the pattern of the PSD under the peak frequency was similar to that of the human PSD. However, the frequencies were five times higher in rats than in humans. Based on the analysis of the rats' bipedal standing motion, there were some common characteristics between rat and human standing motions. Thus, using standing rats is expected to be a powerful tool to reveal the neural basis of postural control.

Progressive impairment of cerebellar mGluR signalling and its therapeutic potential for cerebellar ataxia in spinocerebellar ataxia type 1 model mice.

KEY POINTS: Spinocerebellar ataxia type 1 (SCA1) is a progressive neurodegenerative disease caused by a gene defect, leading to movement disorder such as cerebellar ataxia. It remains largely unknown which functional defect contributes to the cerebellar ataxic phenotype in SCA1. In this study, we report progressive dysfunction of metabotropic glutamate receptor (mGluR) signalling, which leads to smaller slow synaptic responses, reduced dendritic Ca2+ signals and impaired synaptic plasticity at cerebellar synapses, in the early disease stage of SCA1 model mice. We also show that enhancement of mGluR signalling by a clinically available drug, baclofen, leads to improvement of motor performance in SCA1 mice. SCA1 is an incurable disease with no effective treatment, and our results may provide mechanistic grounds for targeting mGluRs and a novel drug therapy with baclofen to treat SCA1 patients in the future. ABSTRACT: Spinocerebellar ataxia type 1 (SCA1) is a progressive neurodegenerative disease that presents with cerebellar ataxia and motor learning defects. Previous studies have indicated that the pathology of SCA1, as well as other ataxic diseases, is related to signalling pathways mediated by the metabotropic glutamate receptor type 1 (mGluR1), which is indispensable for proper motor coordination and learning. However, the functional contribution of mGluR signalling to SCA1 pathology is unclear. In the present study, we show that SCA1 model mice develop a functional impairment of mGluR signalling which mediates slow synaptic responses, dendritic Ca2+ signals, and short- and long-term synaptic plasticity at parallel fibre (PF)-Purkinje cell (PC) synapses in a progressive manner from the early disease stage (5 postnatal weeks) prior to PC death. Notably, impairment of mGluR-mediated dendritic Ca2+ signals linearly correlated with a reduction of PC capacitance (cell surface area) in disease progression. Enhancement of mGluR signalling by baclofen, a clinically available GABAB receptor agonist, led to an improvement of motor performance in SCA1 mice and the improvement lasted ∼1 week after a single application of baclofen. Moreover, the restoration of motor performance in baclofen-treated SCA1 mice matched the functional recovery of mGluR-mediated slow synaptic currents and mGluR-dependent short- and long-term synaptic plasticity. These results suggest that impairment of synaptic mGluR cascades is one of the important contributing factors to cerebellar ataxia in early and middle stages of SCA1 pathology, and that modulation of mGluR signalling by baclofen or other clinical interventions may be therapeutic targets to treat SCA1.

Involvement of GluD2 in Fear-Conditioned Bradycardia in Mice.

Lesions in the cerebellar vermis abolish acquisition of fear-conditioned bradycardia in animals and human patients. The δ2 glutamate receptor (GluD2) is predominantly expressed in cerebellar Purkinje cells. The mouse mutant ho15J carries a spontaneous mutation in GluD2 and these mice show a primary deficiency in parallel fiber-Purkinje cell synapses, multiple innervations of Purkinje cells by climbing fibers, and impairment of long-term depression. In the present study, we used ho15J mice to investigate the role of the cerebellum in fear-conditioned bradycardia. We recorded changes in heart rate of ho15J mice induced by repeated pairing of an acoustic (conditioned) stimulus (CS) with an aversive (unconditioned) stimulus (US). The mice acquired conditioned bradycardia on Day 1 of the CS-US phase, similarly to wild-type mice. However, the magnitude of the conditioned bradycardia was not stable in the mutant mice, but rather was exaggerated on Days 2-5 of the CS-US phase. We examined the effects of reversibly inactivating the cerebellum by injection of an antagonist against the α-amino-3-hydroxy-5-methyl-4-isoxazole propionate receptor (AMPAR). The antagonist abolished expression of conditioned responses in both wild-type and ho15J mice. We conclude that the GluD2 mutation in the ho15J mice affects stable retention of the acquired conditioned bradycardia.

Exercise-induced expression of monocarboxylate transporter 2 in the cerebellum and its contribution to motor performance.

Monocarboxylate transporter 2 (MCT2) is an important component of the lactate transport system in neurons of the adult brain. Purkinje cells in the cerebellum have been shown to have high levels of MCT2, suggesting that this protein has a key function in energy metabolism and neuronal activities in these cells. However, it is not known whether inhibition of lactate transport via MCT2 in the cerebellum affects motor performance. To address this question, we examined motor performance in mice following the inhibition of lactate transport via MCT2 in the cerebellum using α-cyano-4-hydroxycinnamate (4-CIN). 4-CIN or saline was injected into the subarachnoidal space of the cerebellum of mice and motor performance was analyzed by a rotarod test both before and after injection. 4-CIN injection reduced retention time in the rotarod test by approximately 80% at 1h post-injection compared with pre-injection. No effect was observed at 2h post-injection or in mice treated with the vehicle control. Because we observed that MCT2 plays an important role in motor performance, we next investigated the effects of acute exercise on MCT2 transcription and protein levels in mice sampled pre-exercise and at 0 and 5h after 2h of treadmill running. We found a significant increase in MCT2 mRNA levels, but not of protein levels, in the cerebellum at 5h after exercise. Our results indicate that lactate transport via MCT2 in the cerebellum may play an important role in motor performance and that exercise can increase MCT2 expression at the transcriptional level.

Aß induces oxidative stress in senescence-accelerated (SAMP8) mice.

According to the amyloid hypothesis, amyloid ß accumulates in brains with Alzheimer's disease (AD) and triggers cell death and memory deficit. Previously, we developed a rice Aß vaccine expressing Aß, which reduced brain Aß levels in the Tg2576 mouse model of familial AD. We used senescence-accelerated SAMP8 mice as a model of sporadic AD and investigated the relationship between Aß and oxidative stress. Insoluble Aß and 4-hydroxynonenal (4-HNE) levels tended to be reduced in SAMP8 mice-fed the rice Aß vaccine. We attempted to clarify the relationship between oxidative stress and Aß in vitro. Addition of Aß peptide to the culture medium resulted in an increase in 4-HNE levels in SH-SY5Y cells. Tg2576 mice, which express large amounts of Aß in their brain, also exhibited increased 4-HNE levels; this increase was inhibited by the Aß vaccine. These results indicate that Aß induces oxidative stress in cultured cells and in the mouse brain.

Measuring body sway of bipedally standing rat and quantitative evaluation of its postural control.

Human generates very slow (<1 Hz) body sway during standing, and the behavior of this sway is known to be changed characteristically depending on the neural ataxia. In order to investigate the sway mechanism and mechanism of neural ataxia through this sway behavior, the present research proposes an experimental environment of rats under bipedal standing. By the experiment, we succeeded the measurement of six intact rats standing for over 200 seconds without postural supports. Moreover, by comparing measured center of pressure (COP) and that of system model with nonlinear PID control model which is proposed as human standing model, control parameters of rats were numerically evaluated. Evaluated control parameters of rats were close to those of human, i.e., control strategy was considered to be comparable between rats and human.

Investigation of adaptive split-belt treadmill walking by the hindlimbs of rats.

In this study, we investigated the adaptive behavior during hindlimb locomotion of rats on a split-belt treadmill. We measured and analyzed the movement of intact rats walking by the hindlimbs on the splitbelt treadmill with two conditions: symmetric and asymmetric belt speed. In addition, we conducted the dynamic simulation of a neuromusculoskeletal model of rat's hindlimb walking on a split-belt treadmill. We investigated the immediate modulations of the duty factors and relative phase between the right and left limbs depending on the conditions of the treadmill. The results of the simulation were qualitatively similar to those of the measurement experiment. Furthermore, these results were qualitatively similar to the measurement data of the humans and cats in the previous studies. This suggests that our model have the essential aspects to produce the adaptive split-belt treadmill walking in dynamics viewpoints.

Involvement of cholinergic system in hyperactivity in dopamine-deficient mice.

Dopaminergic systems have been known to be involved in the regulation of locomotor activity and development of psychosis. However, the observations that some Parkinson's disease patients can move effectively under appropriate conditions despite low dopamine levels (eg, kinesia paradoxia) and that several psychotic symptoms are typical antipsychotic resistant and atypical antipsychotic sensitive indicate that other systems beyond the dopaminergic system may also affect locomotor activity and psychosis. The present study showed that dopamine-deficient (DD) mice, which had received daily L-DOPA injections, could move effectively and even be hyperactive 72 h after the last L-DOPA injection when dopamine was almost completely depleted. Such hyperactivity was ameliorated by clozapine but not haloperidol or ziprasidone. Among multiple actions of clozapine, muscarinic acetylcholine (ACh) activation markedly reduced locomotor activity in DD mice. Furthermore, the expression of choline acetyltransferase, an ACh synthase, was reduced and extracellular ACh levels were significantly reduced in DD mice. These results suggest that the cholinergic system, in addition to the dopaminergic system, may be involved in motor control, including hyperactivity and psychosis. The present findings provide additional evidence that the cholinergic system may be targeted for the treatment of Parkinson's disease and psychosis.