Localized chemogenetic silencing of inhibitory neurons: a novel mouse model of focal cortical epileptic activity.

Focal cortical epilepsies are frequently refractory to available anticonvulsant drug therapies. One key factor contributing to this state is the limited availability of animal models that allow to reliably study focal cortical seizures and how they recruit surrounding brain areas in vivo. In this study, we selectively expressed the inhibitory chemogenetic receptor, hM4D, in GABAergic neurons in focal cortical areas using viral gene transfer. GABAergic silencing using Clozapine-N-Oxide (CNO) demonstrated reliable induction of local epileptiform events in the electroencephalogram signal of awake freely moving mice. Anesthetized mice experiments showed consistent induction of focal epileptiform-events in both the barrel cortex (BC) and the medial prefrontal cortex (mPFC), accompanied by high-frequency oscillations, a known characteristic of human seizures. Epileptiform-events showed propagation indication with favored propagation pathways: from the BC on 1 hemisphere to its counterpart and from the BC to the mPFC, but not vice-versa. Lastly, sensory whisker-pad stimulation evoked BC epileptiform events post-CNO, highlighting the potential use of this model in studying sensory-evoked seizures. Combined, our results show that targeted chemogenetic inhibition of GABAergic neurons using hM4D can serve as a novel, versatile, and reliable model of focal cortical epileptic activity suitable for systematically studying cortical ictogenesis in different cortical areas.

Area-specific thalamocortical synchronization underlies the transition from motor planning to execution.

We studied correlated firing between motor thalamic and cortical cells in monkeys performing a delayed-response reaching task. Simultaneous recording of thalamocortical activity revealed that around movement onset, thalamic cells were positively correlated with cell activity in the primary motor cortex but negatively correlated with the activity of the premotor cortex. The differences in the correlation contrasted with the average neural responses, which were similar in all three areas. Neuronal correlations reveal functional cooperation and opposition between the motor thalamus and distinct motor cortical areas with specific roles in planning vs. performing movements. Thus, by enhancing and suppressing motor and premotor firing, the motor thalamus can facilitate the transition from a motor plan to execution.

Ketamine induced converged synchronous gamma oscillations in the cortico-basal ganglia network of nonhuman primates.

N-methyl-d-aspartate (NMDA) antagonists are widely used in anesthesia, pain management, and schizophrenia animal model studies, and recently as potential antidepressants. However, the mechanisms underlying their anesthetic, psychotic, cognitive, and emotional effects are still elusive. The basal ganglia (BG) integrate input from different cortical domains through their dopamine-modulated connections to achieve optimal behavior control. NMDA antagonists have been shown to induce gamma oscillations in human EEG recordings and in rodent cortical and BG networks. However, network relations and implications to the primate brain are still unclear. We recorded local field potentials (LFPs) simultaneously from the primary motor cortex (M1) and the external globus pallidus (GPe) of four vervet monkeys (26 sessions, 97 and 76 cortical and pallidal LFPs, respectively) before and after administration of ketamine (NMDA antagonist, 10 mg/kg im). Ketamine induced robust, spontaneous gamma (30-50 Hz) oscillations in M1 and GPe. These oscillations were initially modulated by ultraslow oscillations (~0.3 Hz) and were highly synchronized within and between M1 and the GPe (mean coherence magnitude = 0.76, 0.88, and 0.41 for M1-M1, GPe-GPe, and M1-GPe pairs). Phase differences were distributed evenly around zero with broad and very narrow distribution for the M1-M1 and GPe-GPe pairs (-3.5 ± 31.8° and -0.4 ± 6.0°), respectively. The distribution of M1-GPe phase shift was skewed to the left with a mean of -18.4 ± 20.9°. The increased gamma coherence between M1 and GPe, two central stages in the cortico-BG loops, suggests a global abnormal network phenomenon with a unique spectral signature, which is enabled by the BG funneling architecture.NEW & NOTEWORTHY This study is the first to show spontaneous gamma oscillations under NMDA antagonist in nonhuman primates. These oscillations appear in synchrony in the cortex and the basal ganglia. Phase analysis refutes the confounding effects of volume conduction and supports the funneling and amplifying architecture of the cortico-basal ganglia loops. These results suggest an abnormal network phenomenon with a unique spectral signature that could account for pathological mental and neurological states.

Neurons in both pallidal segments change their firing properties similarly prior to closure of the eyes.

Current anatomical models of the cortico-basal ganglia (BG) network predict reciprocal discharge patterns between the external and internal segments of the globus pallidus (GPe and GPi, respectively), as well as cortical driving of BG activity. However, physiological studies revealing similarity in the transient responses of GPe and GPi neurons cast doubts on these predictions. Here, we studied the discharge properties of GPe, GPi, and primary motor cortex neurons of two monkeys in two distinct states: when eyes are open versus when they are closed. Both pallidal populations exhibited decreased discharge rates in the "eye closed" state accompanied by elevated values of the coefficient of variation (CV) of their interspike interval (ISI) distributions. The pallidal modulations in discharge patterns were partially attributable to larger fractions of longer ISIs in the "eye closed" state. In addition, the pallidal discharge modulations were gradual, starting prior to closing of the eyes. Cortical neurons, as opposed to pallidal neurons, increased their discharge rates steeply on closure of the eyes. Surprisingly, the cortical rate modulations occurred after pallidal modulations. However, as in the pallidum, the CV values of cortical ISI distributions increased in the "eye closed" state, indicating a more bursty discharge pattern in that state. Thus changes in GPe and GPi discharge properties were positively correlated, suggesting that the subthalamic nucleus and/or the striatum constitute the main common driving force for both pallidal segments. Furthermore, the early, unexpected changes in the pallidum are better explained by a subcortical rather than a cortical loop through the BG.

Midbrain dopaminergic neurons and striatal cholinergic interneurons encode the difference between reward and aversive events at different epochs of probabilistic classical conditioning trials.

Midbrain dopaminergic neurons (DANs) typically increase their discharge rate in response to appetitive predictive cues and outcomes, whereas striatal cholinergic tonically active interneurons (TANs) decrease their rate. This may indicate that the activity of TANs and DANs is negatively correlated and that TANs can broaden the basal ganglia reinforcement teaching signal, for instance by encoding worse than predicted events. We studied the activity of 106 DANs and 180 TANs of two monkeys recorded during the performance of a classical conditioning task with cues predicting the probability of food, neutral, and air puff outcomes. DANs responded to all cues with elevations of discharge rate, whereas TANs depressed their discharge rate. Nevertheless, although dopaminergic responses to appetitive cues were larger than their responses to neutral or aversive cues, the TAN responses were more similar. Both TANs and DANs responded faster to an air puff than to a food outcome; however, DANs responded with a discharge elevation, whereas the TAN responses included major negative and positive deflections. Finally, food versus air puff omission was better encoded by TANs. In terms of the activity of single neurons with distinct responses to the different behavioral events, both DANs and TANs were more strongly modulated by reward than by aversive related events and better reflected the probability of reward than aversive outcome. Thus, TANs and DANs encode the task episodes differentially. The DANs encode mainly the cue and outcome delivery, whereas the TANs mainly encode outcome delivery and omission at termination of the behavioral trial episode.

An assumption-free quantification of neural responses to electrical stimulations.

BACKGROUND: Connectivity between brain regions provides the fundamental infrastructure for information processing. The standard way to characterize these interactions is to stimulate one site while recording the evoked response from a second site. The average stimulus-triggered response is usually compared to the pre-stimulus activity. This requires a set of prior assumptions regarding the amplitude and duration of the evoked response. NEW METHOD: We introduce an assumption-free method for detecting and clustering evoked responses. We used Independent Component Analysis to reduce the dimensions of the response vectors, and then clustered them according to a Gaussian mixture model. This enables both the detection and categorization of responsive sites into different subtypes. RESULTS: Our method is demonstrated on recordings obtained from the sensory-motor cortex of behaving primates in response to stimulation of the cerebello-thalamo-cortical tract. We detected and classified the evoked responses of local field potential (LFP) and local spiking activity (multiunit activity-MUA). We found a strong association between specific input (LFP) and output (MUA) patterns across cortical sites, further supporting the physiological relevance of the proposed method. COMPARISON WITH EXISTING METHODS: Our method detected the vast majority of sites found in the conventional, significant threshold-crossing method. However, we found a subgroup of sites with a robust response that were missed when using the conventional method. CONCLUSION: Our method provides a useful, assumption-free tool for detecting and classifying neural evoked responses in a physiologically-relevant manner.

Closed-loop deep brain stimulation is superior in ameliorating parkinsonism.

Continuous high-frequency deep brain stimulation (DBS) is a widely used therapy for advanced Parkinson's disease (PD) management. However, the mechanisms underlying DBS effects remain enigmatic and are the subject of an ongoing debate. Here, we present and test a closed-loop stimulation strategy for PD in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) primate model of PD. Application of pallidal closed-loop stimulation leads to dissociation between changes in basal ganglia (BG) discharge rates and patterns, providing insights into PD pathophysiology. Furthermore, cortico-pallidal closed-loop stimulation has a significantly greater effect on akinesia and on cortical and pallidal discharge patterns than standard open-loop DBS and matched control stimulation paradigms. Thus, closed-loop DBS paradigms, by modulating pathological oscillatory activity rather than the discharge rate of the BG-cortical networks, may afford more effective management of advanced PD. Such strategies have the potential to be effective in additional brain disorders in which a pathological neuronal discharge pattern can be recognized.

Neighboring Pallidal Neurons Do Not Exhibit more Synchronous Oscillations than Remote Ones in the MPTP Primate Model of Parkinson's Disease.

In the healthy primate, neurons of the external and internal segments of the globus pallidus (GP) present a primarily irregular firing pattern, and a negligible level of synchrony is observed between pairs of neurons. This holds even for neighboring cells, despite their higher probability to receive common inputs and to innervate each other via lateral connectivity. In the Parkinsonian primate, this changes drastically, and many pairs of GP cells show synchronous oscillations. To address the relation between distance and synchrony in the Parkinsonian state, we compared the synchrony of discharge of close pairs of neurons, recorded by the same electrode, with remote pairs, recorded by different ones. However, spike trains of neighboring cells recorded by the same extracellular electrode exhibit the shadowing effect; i.e., lack of detection of spikes that occur within a few milliseconds of each other. Here, we demonstrate that the shadowing artifact can both induce apparent correlations between non-correlated neurons, as well as conceal existing correlations between neighboring ones. We therefore introduced artificial shadowing in the remote pairs, similar to the effect we observed in the close ones. After the artificial shadowing, neighboring cells did not show a higher tendency to oscillate synchronously than remote ones. On the contrary, the average percentage (over all sessions) of artificially shadowed remote pairs exhibiting synchronous oscillations was 35.4% compared to 17.2% in the close ones. Similar trend was found when the unshadowed remote pairs were separated according to the estimated distance between electrode tips: 29.9% of pairs at approximate distance of less than 750 µm were significantly synchronized, in comparison with 28.5% of the pairs whose distance was more than 750 µm. We conclude that the synchronous oscillations in the GP of MPTP treated primates are homogenously distributed.

A noninvasive, fast and inexpensive tool for the detection of eye open/closed state in primates.

Accurate detection of the eye state (i.e., open or closed) of animals during electrophysiological recordings is often crucial for analyzing physiological data. This requires a system which is reliable, and preferably noninvasive and inexpensive. Here we present such a tool incorporating a standard digital camera and a semi-automatic eye state detection (ESD) algorithm that can be used easily in typical primate electrophysiological setups. The ESD algorithm is based on the high light absorbance of the iris and pupil relative to the eyelid and takes advantage of the unique conditions found in primate physiological recordings (minimal area of sclera and head fixation). The ESD algorithm is as accurate as a human observer, and is not vulnerable to variance inherent to human decisions that it requires (i.e., eye location setting, training set classification and threshold setting). The temporal resolution with standard interlaced digital cameras is 17-20 ms. This is sufficient for the detection of eye state changes during electrophysiological recordings including spontaneous blinking and eye blink conditioning, as demonstrated here. Furthermore, the ESD tool can be applied to other physiological areas of research in which changes in eye state are critical to analyzing neuronal activity.

Analyzing octopus movements using three-dimensional reconstruction.

Octopus arms, as well as other muscular hydrostats, are characterized by a very large number of degrees of freedom and a rich motion repertoire. Over the years, several attempts have been made to elucidate the interplay between the biomechanics of these organs and their control systems. Recent developments in electrophysiological recordings from both the arms and brains of behaving octopuses mark significant progress in this direction. The next stage is relating these recordings to the octopus arm movements, which requires an accurate and reliable method of movement description and analysis. Here we describe a semiautomatic computerized system for 3D reconstruction of an octopus arm during motion. It consists of two digital video cameras and a PC computer running custom-made software. The system overcomes the difficulty of extracting the motion of smooth, nonrigid objects in poor viewing conditions. Some of the trouble is explained by the problem of light refraction in recording underwater motion. Here we use both experiments and simulations to analyze the refraction problem and show that accurate reconstruction is possible. We have used this system successfully to reconstruct different types of octopus arm movements, such as reaching and bend initiation movements. Our system is noninvasive and does not require attaching any artificial markers to the octopus arm. It may therefore be of more general use in reconstructing other nonrigid, elongated objects in motion.