Neurons in the Nonhuman Primate Amygdala and Dorsal Anterior Cingulate Cortex Signal Aversive Memory Formation under Sedation.

BACKGROUND: Anesthetics aim to prevent memory of unpleasant experiences. The amygdala and dorsal anterior cingulate cortex participate in forging emotional and valence-driven memory formation. It was hypothesized that this circuitry maintains its role under sedation. METHODS: Two nonhuman primates underwent aversive tone-odor conditioning under sedative states induced by ketamine or midazolam (1 to 8 and 0.1 to 0.8 mg/kg, respectively). The primary outcome was behavioral and neural evidence suggesting memory formation. This study simultaneously measured conditioned inspiratory changes and changes in firing rate of single neurons in the amygdala and the dorsal anterior cingulate cortex in response to an expected aversive olfactory stimulus appearing during acquisition and tested their retention after recovery. RESULTS: Aversive memory formation occurred in 26 of 59 sessions under anesthetics (16 of 29 and 10 of 30, 5 of 30 and 21 of 29 for midazolam and ketamine at low and high doses, respectively). Single-neuron responses in the amygdala and dorsal anterior cingulate cortex were positively correlated between acquisition and retention (amygdala, n = 101, r = 0.51, P < 0.001; dorsal anterior cingulate cortex, n = 121, r = 0.32, P < 0.001). Neural responses during acquisition under anesthetics were stronger in sessions exhibiting memory formation than those that did not (amygdala median response ratio, 0.52 versus 0.33, n = 101, P = 0.021; dorsal anterior cingulate cortex median response ratio, 0.48 versus 0.32, n = 121, P = 0.012). The change in firing rate of amygdala neurons during acquisition was correlated with the size of stimuli-conditioned inspiratory response during retention (n = 101, r = 0.22 P = 0.026). Thus, amygdala and dorsal anterior cingulate cortex responses during acquisition under anesthetics predicted retention. Respiratory unconditioned responses to the aversive odor anesthetics did not differ from saline controls. CONCLUSIONS: These results suggest that the amygdala-dorsal anterior cingulate cortex circuit maintains its role in acquisition and maintenance of aversive memories in nonhuman primates under sedation with ketamine and midazolam and that the stimulus valence is sufficient to drive memory formation.

Shared yet dissociable neural codes across eye gaze, valence and expectation.

The direction of the eye gaze of others is a prominent social cue in primates and is important for communication1-11. Although gaze can signal threat and elicit anxiety6,12,13, it remains unclear whether it shares neural circuitry with stimulus value. Notably, gaze not only has valence, but can also serve as a predictor of the outcome of a social encounter, which can be either negative or positive2,8,12,13. Here we show that the neural codes for gaze and valence overlap in primates and that they involve two different mechanisms: one for the outcome and another for its expectation. Monkeys participated in the human intruder test13,14, in which a human participant had either a direct or averted gaze, interleaved with blocks of aversive and appetitive conditioning. We find that single neurons in the amygdala encode gaze15, whereas neurons in the anterior cingulate cortex encode the social context16, but not gaze. We identify a shared population in the amygdala for which the neural responses to direct and averted gaze parallel the responses to aversive and appetitive stimulus, respectively. Furthermore, we distinguish between two neural mechanisms-an overall-activity scheme that is used for gaze and the unconditioned stimulus, and a correlated-selectivity scheme that is used for gaze and the conditioned stimulus. These findings provide insights into the origins of the neural mechanisms that underlie the computations of both social interactions and valence, and could help to shed light on mechanisms that underlie social anxiety and the comorbidity between anxiety and impaired social interactions.

Long time-scales in primate amygdala neurons support aversive learning.

Associative learning forms when there is temporal relationship between a stimulus and a reinforcer, yet the inter-trial-interval (ITI), which is usually much longer than the stimulus-reinforcer-interval, contributes to learning-rate and memory strength. The neural mechanisms that enable maintenance of time between trials remain unknown, and it is unclear if the amygdala can support time scales at the order of dozens of seconds. We show that the ITI indeed modulates rate and strength of aversive-learning, and that single-units in the primate amygdala and dorsal-anterior-cingulate-cortex signal confined periods within the ITI, strengthen this coding during acquisition of aversive-associations, and diminish during extinction. Additionally, pairs of amygdala-cingulate neurons synchronize during specific periods suggesting a shared circuit that maintains the long temporal gap. The results extend the known roles of this circuit and suggest a mechanism that maintains trial-structure and temporal-contingencies for learning.

Oscillations Synchronize Amygdala-to-Prefrontal Primate Circuits during Aversive Learning.

The contribution of oscillatory synchrony in the primate amygdala-prefrontal pathway to aversive learning remains largely unknown. We found increased power and phase synchrony in the theta range during aversive conditioning. The synchrony was linked to single-unit spiking and exhibited specific directionality between input and output measures in each region. Although it was correlated with the magnitude of conditioned responses, it declined once the association stabilized. The results suggest that amygdala spikes help to synchronize ACC activity and transfer error signal information to support memory formation.

A neuro-inspired model-based closed-loop neuroprosthesis for the substitution of a cerebellar learning function in anesthetized rats.

Neuroprostheses could potentially recover functions lost due to neural damage. Typical neuroprostheses connect an intact brain with the external environment, thus replacing damaged sensory or motor pathways. Recently, closed-loop neuroprostheses, bidirectionally interfaced with the brain, have begun to emerge, offering an opportunity to substitute malfunctioning brain structures. In this proof-of-concept study, we demonstrate a neuro-inspired model-based approach to neuroprostheses. A VLSI chip was designed to implement essential cerebellar synaptic plasticity rules, and was interfaced with cerebellar input and output nuclei in real time, thus reproducing cerebellum-dependent learning in anesthetized rats. Such a model-based approach does not require prior system identification, allowing for de novo experience-based learning in the brain-chip hybrid, with potential clinical advantages and limitations when compared to existing parametric "black box" models.

A Cerebellar Neuroprosthetic System: Computational Architecture and in vivo Test.

Emulating the input-output functions performed by a brain structure opens the possibility for developing neuroprosthetic systems that replace damaged neuronal circuits. Here, we demonstrate the feasibility of this approach by replacing the cerebellar circuit responsible for the acquisition and extinction of motor memories. Specifically, we show that a rat can undergo acquisition, retention, and extinction of the eye-blink reflex even though the biological circuit responsible for this task has been chemically inactivated via anesthesia. This is achieved by first developing a computational model of the cerebellar microcircuit involved in the acquisition of conditioned reflexes and training it with synthetic data generated based on physiological recordings. Secondly, the cerebellar model is interfaced with the brain of an anesthetized rat, connecting the model's inputs and outputs to afferent and efferent cerebellar structures. As a result, we show that the anesthetized rat, equipped with our neuroprosthetic system, can be classically conditioned to the acquisition of an eye-blink response. However, non-stationarities in the recorded biological signals limit the performance of the cerebellar model. Thus, we introduce an updated cerebellar model and validate it with physiological recordings showing that learning becomes stable and reliable. The resulting system represents an important step toward replacing lost functions of the central nervous system via neuroprosthetics, obtained by integrating a synthetic circuit with the afferent and efferent pathways of a damaged brain region. These results also embody an early example of science-based medicine, where on the one hand the neuroprosthetic system directly validates a theory of cerebellar learning that informed the design of the system, and on the other one it takes a step toward the development of neuro-prostheses that could recover lost learning functions in animals and, in the longer term, humans.

Cortical balance of excitation and inhibition is regulated by the rate of synaptic activity.

Cortical activity is determined by the balance between excitation and inhibition. To examine how shifts in brain activity affect this balance, we recorded spontaneous excitatory and inhibitory synaptic inputs into layer 4 neurons from rat somatosensory cortex while altering the depth of anesthesia. The rate of excitatory and inhibitory events was reduced by ∼50% when anesthesia was deepened. However, whereas both the amplitude and width of inhibitory synaptic events profoundly increased under deep anesthesia, those of excitatory events were unaffected. These effects were found using three different types of anesthetics, suggesting that they are caused by the network state and not by local specific action of the anesthetics. To test our hypothesis that the size of inhibitory events increased because of the decreased rate of synaptic activity under deep anesthesia, we blocked cortical excitation and replayed the slow and fast patterns of inhibitory inputs using intracortical electrical stimulation. Evoked inhibition was larger under low-frequency stimulation, and, importantly, this change occurred regardless of the depth of anesthesia. Hence, shifts in the balance between excitation and inhibition across distinct states of cortical activity can be explained by the rate of inhibitory inputs combined with their short-term plasticity properties, regardless of the actual global brain activity.

A VLSI field-programmable mixed-signal array to perform neural signal processing and neural modeling in a prosthetic system.

A very-large-scale integration field-programmable mixed-signal array specialized for neural signal processing and neural modeling has been designed. This has been fabricated as a core on a chip prototype intended for use in an implantable closed-loop prosthetic system aimed at rehabilitation of the learning of a discrete motor response. The chosen experimental context is cerebellar classical conditioning of the eye-blink response. The programmable system is based on the intimate mixing of switched capacitor analog techniques with low speed digital computation; power saving innovations within this framework are presented. The utility of the system is demonstrated by the implementation of a motor classical conditioning model applied to eye-blink conditioning in real time with associated neural signal processing. Paired conditioned and unconditioned stimuli were repeatedly presented to an anesthetized rat and recordings were taken simultaneously from two precerebellar nuclei. These paired stimuli were detected in real time from this multichannel data. This resulted in the acquisition of a trigger for a well-timed conditioned eye-blink response, and repetition of unpaired trials constructed from the same data led to the extinction of the conditioned response trigger, compatible with natural cerebellar learning in awake animals.

Bioactive anti-inflammatory coating for chronic neural electrodes.

Chronic electrodes are widely used for brain degenerative and psychiatric diseases such as Parkinson's disease,major depression, and obsessive-compulsive disorder, and for neuronal prosthesis. Brain immune reaction to electrodes in the form of glial scar encapsulates the electrode and reduces the efficacy of deep brain stimulation and neuronal prosthesis.State-of-the-art strategies for improving brain­electrode interface use passive protein coating to "camouflage" the electrode from the immune system. In this study, we actively reduced the brain immune reaction to the chronic electrodes using immune suppressing protein, that is, interleukin (IL)-1 receptor antagonist. IL-1 receptor antagonist-coated electrodes and non coated electrodes were chronically implanted in rats. An additional group of rats was chronically implanted with IL-1 receptor antagonist- and laminin-coated electrodes (as passive protein). Examination of glial scaring 1 and 4 weeks after implantation indicated a significant reduction in the amount of glial scar in the vicinity of the IL-1 receptor antagonist-coated electrode in comparison to both non coated electrode and laminin-coated electrodes. The results strongly suggest that active immune suppressing protein reduces the level of immune reaction to chronic electrodes already after 1 week after implantation and generates less immune reaction than passive protein coating [corrected].

Amygdala conditioning modulates sensory input to the cerebellum.

Localization of emotional learning in the amygdala and discrete motor learning in the cerebellum provides empirical means to study the mechanisms mediating the interaction between fast emotional and slow motor learning. Behavioral studies have demonstrated that fear conditioning facilitates the motor conditioning. The present study tests the hypothesis that the amygdala output induces this facilitation by increasing the salience of the conditioned stimulus (CS) representation in the pontine nucleus (PN) input to the cerebellum. Paired trials of CS-US (unconditioned stimulus) were applied to anesthetized rats, a condition that allows for amygdala-based fear conditioning but not cerebellar-based motor conditioning. Multiple unit recordings in the PN served to assess the salience of the CS. Results showed that CS-US conditioning increased the PN-reactivity to the CS. Lidocaine-induced reversible inactivation of the amygdala prevented the facilitatory effect of conditioning on the PN-reactivity to the CS. These findings suggest that the amygdala-based conditioned responses reach the PN and increase the salience of the CS signal there, perhaps facilitating cerebellar conditioning. This facilitatory effect of the amygdala may be conceptualized under the 'two-stage theory of learning', which predicts that emotional learning in the first stage accelerates the motor learning in the second stage. We hereby demonstrate the physiological mechanism through which fast emotional learning in the first stage facilitates slow cerebellar learning in the second stage.