Striatal Dopamine Contributions to Skilled Motor Learning.

Coordinated multijoint limb and digit movements-"manual dexterity"-underlie both specialized skills (e.g., playing the piano) and more mundane tasks (e.g., tying shoelaces). Impairments in dexterous skill cause significant disability, as occurs with motor cortical injury, Parkinson's disease, and a range of other pathologies. Clinical observations, as well as basic investigations, suggest that corticostriatal circuits play a critical role in learning and performing dexterous skills. Furthermore, dopaminergic signaling in these regions is implicated in synaptic plasticity and motor learning. Nonetheless, the role of striatal dopamine signaling in skilled motor learning remains poorly understood. Here, we use fiber photometry paired with a genetically encoded dopamine sensor to investigate striatal dopamine release in both male and female mice as they learn and perform a skilled reaching task. Dopamine rapidly increases during a skilled reach and peaks near pellet consumption. In the dorsolateral striatum, dopamine dynamics are faster than in the dorsomedial and ventral striatum. Across training, as reaching performance improves, dopamine signaling shifts from pellet consumption to cues that predict pellet availability, particularly in medial and ventral areas of the striatum. Furthermore, performance prediction errors are present across the striatum, with reduced dopamine release after an unsuccessful reach. These findings show that dopamine dynamics during skilled motor behaviors change with learning and are differentially regulated across striatal subregions.

Sudden Unexpected Death in Epilepsy and Respiratory Defects in a Mouse Model of DEPDC5-Related Epilepsy.

OBJECTIVES: DEPDC5 is a common causative gene in familial focal epilepsy with or without malformations of cortical development. Its pathogenic variants also confer a significantly higher risk for sudden unexpected death in epilepsy (SUDEP), providing opportunities to investigate the pathophysiology intersecting neurodevelopment, epilepsy, and cardiorespiratory function. There is an urgent need to gain a mechanistic understanding of DEPDC5-related epilepsy and SUDEP, identify biomarkers for patients at high risk, and develop preventive interventions. METHODS: Depdc5 was specifically deleted in excitatory or inhibitory neurons in the mouse brain to determine neuronal subtypes that drive epileptogenesis and SUDEP. Electroencephalogram (EEG), cardiac, and respiratory recordings were performed to determine cardiorespiratory phenotypes associated with SUDEP. Baseline respiratory function and the response to hypoxia challenge were also studied in these mice. RESULTS: Depdc5 deletion in excitatory neurons in cortical layer 5 and dentate gyrus caused frequent generalized tonic-clonic seizures and SUDEP in young adult mice, but Depdc5 deletion in cortical interneurons did not. EEG suppression immediately following ictal offset was observed in fatal and non-fatal seizures, but low amplitude rhythmic theta frequency activity was lost only in fatal seizures. In addition, these mice developed baseline respiratory dysfunction prior to SUDEP, during which ictal apnea occurred long before terminal cardiac asystole. INTERPRETATION: Depdc5 deletion in excitatory neurons is sufficient to cause DEPDC5-related epilepsy and SUDEP. Ictal apnea and respiratory dysregulation play critical roles in SUDEP. Our study also provides a novel mouse model to investigate the underlying mechanisms of DEPDC5-related epilepsy and SUDEP. ANN NEUROL 2023;94:812-824.

Distinct Populations of Motor Thalamic Neurons Encode Action Initiation, Action Selection, and Movement Vigor.

Motor thalamus (Mthal) comprises the ventral anterior, ventral lateral, and ventral medial thalamic nuclei in rodents. This subcortical hub receives input from the basal ganglia (BG), cerebellum, and reticular thalamus in addition to connecting reciprocally with motor cortical regions. Despite the central location of Mthal, the mechanisms by which it influences movement remain unclear. To determine its role in generating ballistic, goal-directed movement, we recorded single-unit Mthal activity as male rats performed a two-alternative forced-choice task. A large population of Mthal neurons increased their firing briefly near movement initiation and could be segregated into functional groups based on their behavioral correlates. The activity of "initiation" units was more tightly locked to instructional cues than movement onset, did not predict which direction the rat would move, and was anticorrelated with reaction time (RT). Conversely, the activity of "execution" units was more tightly locked to movement onset than instructional cues, predicted which direction the rat would move, and was anticorrelated with both RT and movement time. These results suggest that Mthal influences choice RT performance in two stages: short latency, nonspecific action initiation followed by action selection/invigoration. We discuss the implications of these results for models of motor control incorporating BG and cerebellar circuits.SIGNIFICANCE STATEMENT Motor thalamus (Mthal) is a central node linking subcortical and cortical motor circuits, though its precise role in motor control is unclear. Here, we define distinct populations of Mthal neurons that either encode movement initiation, or both action selection and movement vigor. These results have important implications for understanding how basal ganglia, cerebellar, and motor cortical signals are integrated. Such an understanding is critical to defining the pathophysiology of a range of BG- and cerebellum-linked movement disorders, as well as refining pharmacologic and neuromodulatory approaches to their treatment.

Striatal dopamine contributions to skilled motor learning.

Coordinated multi-joint limb and digit movements - "manual dexterity" - underlie both specialized skills (e.g., playing the piano) and more mundane tasks (e.g., tying shoelaces). Impairments in dexterous skill cause significant disability, as occurs with motor cortical injury, Parkinson's Disease, and a range of other pathologies. Clinical observations, as well as basic investigations, suggest that cortico-striatal circuits play a critical role in learning and performing dexterous skills. Furthermore, dopaminergic signaling in these regions is implicated in synaptic plasticity and motor learning. Nonetheless, the role of striatal dopamine signaling in skilled motor learning remains poorly understood. Here, we use fiber photometry paired with a genetically encoded dopamine sensor to investigate striatal dopamine release as mice learn and perform a skilled reaching task. Dopamine rapidly increases during a skilled reach and peaks near pellet consumption. In dorsolateral striatum, dopamine dynamics are faster than in dorsomedial and ventral striatum. Across training, as reaching performance improves, dopamine signaling shifts from pellet consumption to cues that predict pellet availability, particularly in medial and ventral areas of striatum. Furthermore, performance prediction errors are present across the striatum, with reduced dopamine release after an unsuccessful reach. These findings show that dopamine dynamics during skilled motor behaviors change with learning and are differentially regulated across striatal subregions.

Selective inhibition of striatal fast-spiking interneurons causes dyskinesias.

Fast-spiking interneurons (FSIs) can exert powerful control over striatal output, and deficits in this cell population have been observed in human patients with Tourette syndrome and rodent models of dystonia. However, a direct experimental test of striatal FSI involvement in motor control has never been performed. We applied a novel pharmacological approach to examine the behavioral consequences of selective FSI suppression in mouse striatum. IEM-1460, an inhibitor of GluA2-lacking AMPARs, selectively blocked synaptic excitation of FSIs but not striatal projection neurons. Infusion of IEM-1460 into the sensorimotor striatum reduced the firing rate of FSIs but not other cell populations, and elicited robust dystonia-like impairments. These results provide direct evidence that hypofunction of striatal FSIs can produce movement abnormalities, and suggest that they may represent a novel therapeutic target for the treatment of hyperkinetic movement disorders.

Dopamine neuron stimulation induces context-dependent abnormal involuntary movements in healthy rats.

With continued levodopa treatment, most patients with Parkinson disease (PD) develop levodopa-induced dyskinesias (LIDs)-abnormal involuntary movements (AIMs) characterized primarily by chorea. Clinically, LIDs depend on nigrostriatal degeneration and sensitization to repeated levodopa doses. However, the degree of dopamine denervation is correlated with levodopa-induced changes in striatal dopamine. Therefore, pulsatile dopamine release may induce AIMs independently of nigrostriatal degeneration. We optogenetically stimulated dopamine neurons in healthy rats as they engaged in skilled reaching. Repeated stimulation induced progressive AIMs whose severity was modified by behavioral context. AIMs were milder with stimulation during reaches, and more severe if stimulation occurred between reaches. Despite gradual induction, AIMs recurred immediately with subsequent dopamine neuron stimulation. Thus, nigrostriatal denervation is not necessary for fluctuating striatal dopamine to induce AIMs, and behavioral context modulates AIM expression. Furthermore, pulsatile dopamine release induces persistent changes in motor circuits that are revealed by subsequent dopamine neuron activation in appropriate contexts.

A dystonia mouse model with motor and sequencing deficits paralleling human disease.

The dystonias are a group of movement disorders characterized by involuntary twisting movements and postures. A lack of well characterized behavioral models of dystonia has impeded identification of circuit abnormalities giving rise to the disease. Most mouse behavioral assays are implemented independently of cortex, but cortical dysfunction is implicated in human dystonia. It is therefore important to identify dystonia models in which motor cortex-dependent behaviors are altered in ways relevant to human disease. The goal of this study was to characterize a cortically-dependent behavior in the recently-developed Dlx-CKO mouse model of DYT1 dystonia. Mice performed two tasks: skilled reaching and water-elicited grooming. These tests assess motor learning, dexterous skill, and innate motor sequencing. Furthermore, skilled reaching depends strongly on motor cortex, while dorsal striatum is critical for normal grooming. Dlx-CKO mice exhibited significantly lower success rates and pellet contacts compared to control mice during skilled reaching. Despite the skilled reaching impairments, Dlx-CKO mice adapt their reaching strategies. With training, they more consistently contacted the target. Grooming patterns of Dlx-CKO mice are more disorganized than in control mice, as evidenced by a higher proportion of non-chain grooming. However, when Dlx-CKO mice engage in syntactic chains, they execute them similarly to control mice. These abnormalities may provide targets for preclinical intervention trials, as well as facilitate determination of the physiologic path from torsinA dysfunction to motor phenotype.

Evolution of Gross Forelimb and Fine Digit Kinematics during Skilled Reaching Acquisition in Rats.

The ability to learn dexterous motor skills is a fundamental aspect of human behavior. However, the underlying neural circuit mechanisms for dexterous skill learning are unclear. Advancing our understanding of motor skill learning requires the integration of modern neuroscientific techniques with a rigorously characterized dexterous task. The development of automated rodent skilled reaching with paw tracking allows detailed analysis of how reach-to-grasp kinematics evolve during learning. We assessed how both "gross" forelimb and "fine" digit kinematics changed as rats learned skilled reaching. Rats whose success rates increased (learners) consistently reduced the variability in their reach trajectories. Refinement of fine digit control generally continued after consistency in gross hand transport to the pellet plateaued. Interestingly, most rats whose success rates did not increase (non-learners) also converged on consistent reach kinematics. Some non-learners, however, maintained substantial variability in hand and digit trajectories throughout training. These results suggest that gross and fine motor components of dexterous skill are, on average, learned over different timescales. Nonetheless, there is significant intersubject variability in learning rates as assessed by both reaching success and consistency of reach kinematics.

Revisiting the "Paradox of Stereotaxic Surgery": Insights Into Basal Ganglia-Thalamic Interactions.

Basal ganglia dysfunction is implicated in movement disorders including Parkinson Disease, dystonia, and choreiform disorders. Contradicting standard "rate models" of basal ganglia-thalamic interactions, internal pallidotomy improves both hypo- and hyper-kinetic movement disorders. This "paradox of stereotaxic surgery" was recognized shortly after rate models were developed, and is underscored by the outcomes of deep brain stimulation (DBS) for movement disorders. Despite strong evidence that DBS activates local axons, the clinical effects of lesions and DBS are nearly identical. These observations argue against standard models in which GABAergic basal ganglia output gates thalamic activity, and raise the question of how lesions and stimulation can have similar effects. These paradoxes may be resolved by considering thalamocortical loops as primary drivers of motor output. Rather than suppressing or releasing cortex via motor thalamus, the basal ganglia may modulate the timing of thalamic perturbations to cortical activity. Motor cortex exhibits rotational dynamics during movement, allowing the same thalamocortical perturbation to affect motor output differently depending on its timing with respect to the rotational cycle. We review classic and recent studies of basal ganglia, thalamic, and cortical physiology to propose a revised model of basal ganglia-thalamocortical function with implications for basic physiology and neuromodulation.

Interactions Between Motor Thalamic Field Potentials and Single-Unit Spiking Are Correlated With Behavior in Rats.

Field potential (FP) oscillations are believed to coordinate brain activity over large spatiotemporal scales, with specific features (e.g., phase and power) in discrete frequency bands correlated with motor output. Furthermore, complex correlations between oscillations in distinct frequency bands (phase-amplitude, amplitude-amplitude, and phase-phase coupling) are commonly observed. However, the mechanisms underlying FP-behavior correlations and cross-frequency coupling remain unknown. The thalamus plays a central role in generating many circuit-level neural oscillations, and single-unit activity in motor thalamus (Mthal) is correlated with behavioral output. We, therefore, hypothesized that motor thalamic spiking coordinates motor system FPs and underlies FP-behavior correlations. To investigate this possibility, we recorded wideband motor thalamic (Mthal) electrophysiology as healthy rats performed a two-alternative forced-choice task. Delta (1-4 Hz), beta (13-30 Hz), low gamma (30-70 Hz), and high gamma (70-200 Hz) power were strongly modulated by task performance. As in the cortex, the delta phase was correlated with beta/low gamma power and reaction time. Most interestingly, subpopulations of Mthal neurons defined by their relationship to the behavior exhibited distinct relationships with FP features. Specifically, neurons whose activity was correlated with action selection and movement speed were entrained to delta oscillations. Furthermore, changes in their activity anticipated power fluctuations in beta/low gamma bands. These complex relationships suggest mechanisms for commonly observed FP-FP and spike-FP correlations, as well as subcortical influences on motor output.

Precisely timed dopamine signals establish distinct kinematic representations of skilled movements.

Brain dopamine is critical for normal motor control, as evidenced by its importance in Parkinson Disease and related disorders. Current hypotheses are that dopamine influences motor control by 'invigorating' movements and regulating motor learning. Most evidence for these aspects of dopamine function comes from simple tasks (e.g. lever pressing). Therefore, the influence of dopamine on motor skills requiring multi-joint coordination is unknown. To determine the effects of precisely timed dopamine manipulations on the performance of a complex, finely coordinated dexterous skill, we optogenetically stimulated or inhibited midbrain dopamine neurons as rats performed a skilled reaching task. We found that reach kinematics and coordination between gross and fine movements progressively changed with repeated manipulations. However, once established, rats transitioned abruptly between aberrant and baseline reach kinematics in a dopamine-dependent manner. These results suggest that precisely timed dopamine signals have immediate and long-term influences on motor skill performance, distinct from simply 'invigorating' movement.

Chronic histological effects of the flat interface nerve electrode.

The flat interface nerve electrode (FINE) is designed to reshape peripheral nerves into favorable geometries for selective stimulation. Compared to cylindrical geometries, the ovoid geometries created by the FINE allow more space for contact placement. Furthermore, the amount of electrically excitable tissue between stimulating contacts and target axons is reduced. In this study, the nerve response to the presence of the FINE is examined histologically. Three different FINEs were designed to reshape peripheral nerves to different opening heights designated as 'wide' (1.3 mm), 'medium' (0.5 mm) and 'narrow' (0.1 mm) cuffs. Twelve adult cats were implanted with one cuff each (four in total of each design) on their right sciatic nerves. At least 3 months later, the animals were sacrificed and their sciatic nerves were harvested for histological evaluation. Cross-sectional areas and eccentricities (defined as the major axis divided by the minor axis of the closest fit ellipse to a region) of the nerves were measured to assess the degree of reshaping. The wide and medium cuff designs significantly reshaped the nerves compared to control nerves, though there was no significant difference in eccentricity between nerves implanted with wide and medium cuffs. There was extensive deposition of connective tissue in the epineurium of all nerves implanted with cuffs. No significant difference in fiber counts was measured in any of the groups studied. Only nerves implanted with narrow cuffs showed evidence of axonal injury and/or demyelination. These results, coupled with stimulation selectivity measurements made on the same animals, suggest that safe, selective electrodes can be designed with ovoid geometries. Moderate reshaping caused no damage, while extreme reshaping generated mild-to-moderate nerve damage. It might be possible, however, to redesign the cuffs to slowly reshape the nerves.

An automated rat single pellet reaching system with high-speed video capture.

BACKGROUND: Single pellet reaching is an established task for studying fine motor control in which rats reach for, grasp, and eat food pellets in a stereotyped sequence. Most incarnations of this task require constant attention, limiting the number of animals that can be tested and the number of trials per session. Automated versions allow more interventions in more animals, but must be robust and reproducible. NEW METHOD: Our system automatically delivers single reward pellets for rats to grasp with their forepaw. Reaches are detected using real-time computer vision, which triggers video acquisition from multiple angles using mirrors. This allows us to record high-speed (>300 frames per second) video, and trigger interventions (e.g., optogenetics) with high temporal precision. Individual video frames are triggered by digital pulses that can be synchronized with behavior, experimental interventions, or recording devices (e.g., electrophysiology). The system is housed within a soundproof chamber with integrated lighting and ventilation, allowing multiple skilled reaching systems in one room. RESULTS: We show that rats acquire the automated task similarly to manual versions, that the task is robust, and can be synchronized with optogenetic interventions. COMPARISON WITH EXISTING METHODS: Existing skilled reaching protocols require high levels of investigator involvement, or, if ad libitum, do not allow for integration of high-speed, synchronized data collection. CONCLUSION: This task will facilitate the study of motor learning and control by efficiently recording large numbers of skilled movements. It can be adapted for use with modern neurophysiology, which demands high temporal precision.

Chronic measurement of the stimulation selectivity of the flat interface nerve electrode.

The flat interface nerve electrode (FINE) is an attempt to improve the stimulation selectivity of extraneural electrodes. By reshaping peripheral nerves into elliptical cylinders, central fibers are moved closer to the nerve-electrode interface, and additional surface area is created for contact placement. The goals of this study were to test the hypothesis that greater nerve reshaping leads to improved selectivity and to examine the chronic recruitment properties of the FINE. Three FINEs were developed to reshape peripheral nerves to different degrees. Four electrodes of each type were implanted on the sciatic nerves of 12 cats and tested for selectivity over at least three months. There was physiologic evidence of nerve injury in two cats with the tightest cuffs, but the other animals behaved normally. All cuff types were capable of selectively activating branches of the sciatic nerve, as well as groups of fibers within branches. The electrodes that moderately reshaped the nerves demonstrated the most selectivity. Both the selectivity measurements and the recruitment curve characteristics were stable throughout the implant period. From an electrophysiological standpoint, the FINE is a viable alternative for neuroprosthetic devices. A histological analysis of the nerves is under way to evaluate the safety of the FINE.

Dissociable effects of dopamine on learning and performance within sensorimotor striatum.

Striatal dopamine is an important modulator of current behavior, as seen in the rapid and dramatic effects of dopamine replacement therapy in Parkinson Disease (PD). Yet there is also extensive evidence that dopamine acts as a learning signal, modulating synaptic plasticity within striatum to affect future behavior. Disentangling these "performance" and "learning" functions is important for designing effective, long-term PD treatments. We conducted a series of unilateral drug manipulations and dopamine terminal lesions in the dorsolateral striatum of rats highly-trained to perform brief instructed head/neck movements (two-alternative forced choice task). Reaction times and accuracy were measured longitudinally to determine if task behavior changed immediately, progressed over time, and/or persisted after drug withdrawal. Enhanced dopamine signaling with amphetamine caused an immediate, nonprogressive, and bilateral decrease in reaction times (RT). The altered RT distributions were consistent with reduced distance to threshold in the linear approach to threshold with ergodic rate (LATER) model of decision-making. Conversely, the dopamine antagonist flupenthixol caused experience-dependent, persistent changes in RT and accuracy indicative of a "learning" effect. These RT distributions were consistent with a slowed rate of approach to decision threshold. Our results show that dopaminergic signaling makes dissociable contributions to current and future behavior even within a single striatal subregion, and provide important clues for both models of normal decision-making and the design of novel drug therapies in PD.

Mouse models of neurodevelopmental disease of the basal ganglia and associated circuits.

This chapter focuses on neurodevelopmental diseases that are tightly linked to abnormal function of the striatum and connected structures. We begin with an overview of three representative diseases in which striatal dysfunction plays a key role--Tourette syndrome and obsessive-compulsive disorder, Rett's syndrome, and primary dystonia. These diseases highlight distinct etiologies that disrupt striatal integrity and function during development, and showcase the varied clinical manifestations of striatal dysfunction. We then review striatal organization and function, including evidence for striatal roles in online motor control/action selection, reinforcement learning, habit formation, and action sequencing. A key barrier to progress has been the relative lack of animal models of these diseases, though recently there has been considerable progress. We review these efforts, including their relative merits providing insight into disease pathogenesis, disease symptomatology, and basal ganglia function.