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Acetylcholine (ACh) is thought to control arousal, attention, and learning by slowly modulating cortical excitability and plasticity. Recent studies, however, discovered that cholinergic neurons emit precisely timed signals about the aversive outcome at millisecond precision. To investigate the functional relevance of such phasic cholinergic signaling, we manipulated and monitored cholinergic terminals in the mPFC while male mice associated a neutral conditioned stimulus (CS) with mildly aversive eyelid shock (US) over a short temporal gap. Optogenetic inhibition of cholinergic terminals during the US promoted the formation of the CS–US association. On the contrary, optogenetic excitation of cholinergic terminals during the US blocked the association formation. The bidirectional behavioral effects paralleled the corresponding change in the expression of an activity-regulated gene, c-Fos in the mPFC. In contrast, optogenetic inhibition of cholinergic terminals during the CS impaired associative learning, whereas their excitation had marginal effects. In parallel, photometric recording from cholinergic terminals in the mPFC revealed strong innate phasic responses to the US. With subsequent CS–US pairings, cholinergic terminals weakened the responses to the US while developing strong responses to the CS. The across-session changes in the CS- and US-evoked terminal responses were correlated with associative memory strength. These findings suggest that phasic cholinergic signaling in the mPFC exerts opposite effects on aversive associative learning depending on whether it is emitted by the outcome or the cue.SIGNIFICANCE STATEMENTDrugs compensating for the decline of acetylcholine (ACh) are used for cognitive impairment, such as Alzheimer's disease. However, their beneficial effects are limited, demanding new strategies based on better understandings of how ACh modulates cognition. Here, we report that by manipulating ACh signals in the mPFC, we can control the strength of aversive associative learning in mice. Specifically, the suppression of ACh signals during an aversive outcome facilitated its association with a preceding cue. In contrast, the suppression of ACh signals during the cue impaired learning. Considering that this paradigm depends on the brain regions affected in Alzheimer's disease, our findings indicate that precisely timed control of ACh signals is essential to refine ACh-based strategies for cognitive enhancement.

Memory retrieval is thought to depend on the reinstatement of cortical memory representations guided by pattern completion processes in the hippocampus. The lateral entorhinal cortex (LEC) is one of the intermediary regions supporting hippocampal-cortical interactions and houses neurons that prospectively signal past events in a familiar environment. To investigate the functional relevance of the LEC’s activity for cortical reinstatement, we pharmacologically inhibited the LEC and examined its impact on the stability of ensemble firing patterns in one of the LEC’s efferent targets, the medial prefrontal cortex (mPFC). When male rats underwent multiple epochs of identical stimulus sequences in the same environment, the mPFC maintained a stable ensemble firing pattern across repetitions, particularly when the sequence included pairings of neutral and aversive stimuli. With LEC inhibition, the mPFC still formed an ensemble pattern that accurately captured stimuli and their associations within each epoch. However, LEC inhibition markedly disrupted its consistency across the epochs by decreasing the proportion of mPFC neurons that stably maintained firing selectivity for stimulus associations. Thus, the LEC stabilizes cortical representations of learned stimulus associations, thereby facilitating the recovery of the original memory trace without generating a new, redundant trace for familiar experiences. Failure of this process might underlie retrieval deficits in conditions associated with degeneration of the LEC, such as normal aging and Alzheimer’s disease.SIGNIFICANCE STATEMENTTo recall past events, the brain needs to reactivate the activity patterns that occurred during those events. However, such reinstatement of memory traces is not trivial because it goes against the brain’s natural tendency to restructure the activity patterns continuously. We found that dysfunction of a brain region called the lateral entorhinal cortex (LEC) worsened the drift of the brain activity when rats repeatedly underwent the same events in the same room and made them behave as if they had never experienced these events before. Thus, the LEC stabilizes the brain activity to facilitate the recovery of the original memory trace. Failure of this process might underlie memory problems in elderly and Alzheimer’s disease patients with the degenerated LEC.

The lateral entorhinal cortex (LEC) is an essential component of the brain circuitry supporting long-term memory by serving as an interface between the hippocampus and neocortex. Dysfunction of the LEC affects sensory coding in the hippocampus, leading to a view that the LEC provides the hippocampus with highly processed sensory information. It remains unclear, however, how the LEC modulates neural processing in the neocortical regions. To address this point, we pharmacologically inactivated the LEC of male rats during a temporal associative learning task and examined its impact on local network activity in one of the LEC's efferent targets, the prelimbic region of the medial prefrontal cortex (mPFC). Rats were exposed to two neutral stimuli, one of which was paired with an aversive eyelid shock over a short temporal delay. The LEC inhibition reduced the expression of anticipatory blinking responses to the reinforced stimuli without increasing responses to nonreinforced stimuli. In control rats, both the reinforced and nonreinforced stimuli evoked a short-lived, wide-band increase in the prelimbic network activity. With learning, the initial increase of gamma-band activity started to extend into the interval between the reinforced neutral stimulus and the eyelid shock. LEC inhibition attenuated the learning-induced sustained activity, without affecting the initial transient activity. These results suggest that the integrity of LEC is necessary for the formation of temporal stimulus associations and its neural correlates in the mPFC. Given the minimal effects on the innate network responses to sensory stimuli, the LEC appears not to be the main source of sensory inputs to the mPFC; rather it may provide a framework that shapes the mPFC network response to behaviorally relevant cues.

Memories of daily experiences contain incidental details unique to each experience as well as common latent patterns shared with others. Neural representations focusing on the latter aspect can be reinstated by similar new experiences even though their perceptual features do not match the original experiences perfectly. Such flexible memory use allows for faster learning and better decision-making in novel situations. Here, I review evidence from rodent and primate electrophysiological studies to discuss how memory flexibility is implemented in the spiking activity of neuronal ensembles. These findings uncovered innate and learned coding properties and their potential refinement during sleep that support flexible integration and application of memories for better future adaptation.

Prevailing theories posit that the hippocampus rapidly learns stimulus conjunctions during novel experiences, whereas the neocortex learns slowly through subsequent, off-line interaction with the hippocampus. Parallel evidence, however, shows that the medial prefrontal cortex (mPFC; a critical node of the neocortical network supporting long-term memory storage) undergoes rapid modifications of gene expression, synaptic structure, and physiology at the time of encoding. These observations, along with impaired learning with disrupted mPFC, suggest that mPFC neurons may exhibit rapid neural plasticity during novel experiences; however, direct empirical evidence is lacking. We extracellularly recorded action potentials of cells in the prelimbic region of the mPFC, while male rats received a sequence of stimulus presentations for the first time in life. Moment-to-moment tracking of neural ensemble firing patterns revealed that the prelimbic network activity exhibited an abrupt transition within 1 min after the first encounter of an aversive but not neutral stimulus. This network-level change was driven by ∼15% of neurons that immediately elevated their spontaneous firing rates (FRs) and developed firing responses to a neutral stimulus preceding the aversive stimulus within a few instances of their pairings. When a new sensory stimulus was paired with the same aversive stimulus, about half of these neurons generalized firing responses to the new stimulus association. Thus, prelimbic neurons are capable of rapidly forming ensemble codes for novel stimulus associations within minutes. This circuit property may enable the mPFC to rapidly detect and selectively encode the central content of novel experiences.SIGNIFICANCE STATEMENTDuring a new experience, a region of the brain, called the hippocampus, rapidly forms its memory and later instructs another region, called the neocortex, that stores its content. Consistent with this dominant view, cells in the neocortex gradually strengthen the selectivity for the memory content over weeks after novel experiences. However, we still do not know precisely when these cells begin to develop the selectivity. We found that neocortical cells were capable of forming the selectivity for ongoing events within a few minutes of new experiences. This finding provides support for an alternative view that the neocortex works with, but not follows, the hippocampus to form new memories.

The prefrontal cortex has been implicated in various cognitive processes, including working memory, executive control, decision making, and relational learning. One core computational requirement underlying all these processes is the integration of information across time. When rodents and rabbits associate two temporally discontiguous stimuli, some neurons in the medial prefrontal cortex (mPFC) change firing rates in response to the preceding stimulus and sustain the firing rate during the subsequent temporal interval. These firing patterns are thought to serve as a mechanism to buffer the previously presented stimuli and signal the upcoming stimuli; however, how these critical properties are distributed across different neuron types remains unknown. We investigated the firing selectivity of regular-firing, burst-firing, and fast-spiking neurons in the prelimbic region of the mPFC while rats associated two neutral conditioned stimuli (CS) with one aversive stimulus (US). Analyses of firing patterns of individual neurons and neuron ensembles revealed that regular-firing neurons maintained rich information about CS identity and CS-US contingency during intervals separating the CS and US. Moreover, they further strengthened the latter selectivity with repeated conditioning sessions over a month. The selectivity of burst-firing neurons for both stimulus features was weaker than that of regular-firing neurons, indicating the difference in task engagement between two subpopulations of putative excitatory neurons. In contrast, putative inhibitory, fast-spiking neurons showed a stronger selectivity for CS identity than for CS-US contingency, suggesting their potential role in sensory discrimination. These results reveal a fine-scaled functional organization in the prefrontal network supporting the formation of temporal stimulus associations. NEW & NOTEWORTHY To associate stimuli that occurred separately in time, the brain needs to bridge the temporal gap by maintaining what was presented and predicting what would follow. We show that in rat medial prefrontal cortex, the former function is associated with a subpopulation of putative inhibitory neurons, whereas the latter is supported by a subpopulation of putative excitatory neurons. Our results reveal a distinct contribution of these microcircuit components to neural representations of temporal stimulus associations.

Takehara-Nishiuchi

Associative learning restructures the activity of numerous neurons distributed across cortical and subcortical regions. Individual neurons change the rate or timing of spiking patterns in response to environmental stimuli as they become associated with salient outcomes. Recent large-scale activity monitoring in rodents has uncovered that these learning-related changes occur concertedly across groups of neurons within and between brain regions. These changes yield neuronal representations of learned associations in three types of ensemble dynamics: ensemble firing rates, multineuron coactivity, and sequential activity. Here, I review some of the most robust demonstrations of these dynamics in the rodent neocortex and hippocampus and discuss their potential function in memory encoding, consolidation, and retrieval.

The hippocampus rapidly forms associations among ongoing events as they unfold and later instructs the gradual stabilisation of their memory traces in the neocortex. Although this two-stage model of memory consolidation has gained substantial empirical support, parallel evidence from rodent studies suggests that the neocortex, in particular the medial prefrontal cortex, might work in concert with the hippocampus during the encoding of new experiences. This opinion article first summarises findings from behavioural, electrophysiological, and molecular studies in rodents that uncovered immediate changes in synaptic connectivity and neural selectivity in the medial prefrontal cortex during and shortly after novel experiences. Based on these findings, I then propose a model positing that the medial prefrontal cortex and hippocampus might use different strategies to encode information during novel experiences, leading to the parallel formation of complementary memory traces in the two regions. The hippocampus captures moment-to-moment changes in incoming inputs with accurate spatial and temporal contexts, whereas the medial prefrontal cortex may sort the inputs based on their similarity and integrates them over time. These processes of pattern recognition and integration enable the medial prefrontal cortex to, in real time, capture the central content of novel experience and emit relevancy signal that helps to enhance the contrast between the relevant and incidental features of the experience. This hypothesis serves as a framework for future investigations on the potential top-down modulation that the medial prefrontal cortex may exert over the hippocampus to enable the selective, perhaps more intelligent encoding of new information.

Psychological theories posit that the hippocampus rapidly forms associations among ongoing events as they unfold. During a subsequent maturation process, so-called systems memory consolidation, these associations are gradually stabilized within distributed neocortical circuits through close interactions between the hippocampus and neocortex. In the past 50 years, a major effort in neurobiological research has been directed towards translating these descriptive accounts into tangible, biological processes in the brain. Until the early 2000s, most studies exclusively focused on examining whether the hippocampus becomes unnecessary for memory retrieval once the memory has been consolidated. With recent methodological advances, however, the field shifted attention to several other theoretical accounts and began to uncover the genetic, physiological and structural underpinnings of systems memory consolidation at an unprecedented level of precision. Here I review these neurobiological findings in the past 15 years within a framework of six essential predictions extracted from the psychological theories. Genetic approaches have made it possible to tag neurons that were activated during memory encoding and investigate their physiological and genetic profiles as well as reactivation patterns during subsequent retrieval. In parallel, electrophysiological and imaging approaches detected signs of the gradual refinement of memory representations and its underlying hippocampal-neocortical dialogue in millisecond-resolved neural firing patterns, the inter-region coupling of neural activity, across-day stability of neural ensemble activity and functional connectome. This summary represents substantial progress in our understanding of neurobiological mechanisms of systems memory consolidation whilst also identifying several essential remaining questions for future investigations.

Many cognitive processes, such as episodic memory and decision making, rely on the ability to form associations between two events that occur separately in time. The formation of such temporal associations depends on neural representations of three types of information: what has been presented (trace holding), what will follow (temporal expectation), and when the following event will occur (explicit timing). The present review seeks to link these representations with firing patterns of single neurons recorded while rodents and non-human primates associate stimuli, outcomes, and motor responses over time intervals. Across these studies, two distinct firing patterns were observed in the hippocampus, neocortex, and striatum: some neurons change firing rates during or shortly after the stimulus presentation and sustain the firing rate stably or sidlingly during the subsequent intervals (tonic firings). Other neurons transiently change firing rates during a specific moment within the time intervals (phasic firings), and as a group, they form a sequential firing pattern that covers the entire interval. Clever task designs used in some of these studies collectively provide evidence that both tonic and phasic firing responses represent trace holding, temporal expectation, and explicit timing. Subsequently, we applied machine-learning based classification approaches to the two firing patterns within the same dataset collected from rat medial prefrontal cortex during trace eyeblink conditioning. This quantitative analysis revealed that phasic-firing patterns showed greater selectivity for stimulus identity and temporal position than tonic-firing patterns. Our summary illuminates distributed neural representations of temporal association in the forebrain and generates several ideas for future investigations.

Trace eyeblink conditioning is a hippocampus-dependent associative learning paradigm which is impaired in patients with Alzheimer's disease (AD) and animal AD models. Learning in this paradigm accompanies changes in oscillatory activity in forebrain regions, some of which are loci of pathogenic changes in prodromal AD stages. These observations motivated us to examine how cortical event-related potentials (ERPs) during this paradigm are affected by two features of the asymptomatic, AD-related brain abnormality, entorhinal tau accumulation and mild cholinergic deficit. Adult rats received viral overexpression of P301L mutant human tau in the entorhinal cortex, low-dose scopolamine treatment, or both. Electroencephalograms were recorded with epidural electrodes on the surface of the frontal, parietal, and temporal cortices during differential and reversal trace eyeblink conditioning. All rats developed conditioned responses to one of two stimuli (auditory or visual) paired with mild eyelid shock (CS+), but not to the other stimulus presented alone (CS-). They were also able to adjust the response when the stimulus contingency was reversed. With learning, the amplitude of several ERP components in the frontal and temporal cortices came to differentiate the CS+ from CS-. Scopolamine affected the learning-related change in temporal P2 and other learning-unrelated components in three regions. Entorhinal tau overexpression primary affected the amplitude of temporal visual ERPs and learning-unrelated frontal and temporal auditory ERP components. The double manipulation only affected two components of temporal auditory ERPs. Thus, cortical ERPs during differential associative learning are sensitive to asymptomatic brain abnormality associated with AD.

A neural signature of asymptomatic preclinical Alzheimer's disease (AD) is disrupted connectivity between brain regions; however, its underlying mechanisms remain unknown. Here, we tested whether a preclinical pathologic feature, tau aggregation in the entorhinal cortex (EC) is sufficient to disrupt the coordination of local field potentials (LFPs) between its efferent regions. P301L-mutant human tau or green fluorescent protein (GFP) was virally overexpressed in the EC of adult rats. LFPs were recorded from the dorsal hippocampus and prelimbic medial prefrontal cortex while the rats underwent trace eyeblink conditioning where they learned to associate 2 stimuli separated by a short time interval. In GFP-expressing rats, the 2 regions strengthened phase-phase and amplitude-amplitude couplings of theta and gamma oscillations during the interval separating the paired stimuli. Despite normal memory acquisition, this learning-related, inter-region oscillatory coupling was attenuated in the tau-expressing rats while prefrontal phase-amplitude theta-gamma cross-frequency coupling was elevated. Thus, EC tau aggregation caused aberrant long-range circuit activity during associative learning, identifying a culprit for the neural signature of preclinical AD stages.

Some animals learn to fear a situation after observing another individual come to harm, and this learning is influenced by the animals’ social relationship and history. An important but sometimes overlooked factor in studies of observational fear learning is that social context not only affects observers, but may also influence the behavior and communications expressed by those being observed. Here we sought to investigate whether observational fear learning in the degu (Octodon degus) is affected by social familiarity, and the degree to which vocal expressions of alarm or distress contribute. ‘Demonstrator’ degus underwent contextual fear conditioning in the presence of a cagemate or stranger observer. Among the 15 male pairs, observers of familiar demonstrators exhibited higher freezing rates than observers of strangers when returned to the conditioning environment one day later. Observer freezing during testing was, however, also related to the proportion of short- versus long- inter-call-intervals (ICIs) in vocalizations recorded during prior conditioning. In a regression model that included both social relationship and ICI patterns, only the latter was significant. Further investigation of vocalizations, including use of a novel, directed k-means clustering approach, suggested that temporal structure rather than tonal variations may have been responsible for communicating danger. These data offer insight into how different expressions of distress or fear may impact an observer, adding to the complexity of social context effects in studies of empathy and social cognition. The experiments also offer new data on degu alarm calls and a potentially novel methodological approach to complex vocalizations.

A critical function of attention is to support a state of readiness to enhance stimulus detection, independent of stimulus modality. The nucleus basalis magnocellularis (NBM) is the major source of the neurochemical acetylcholine (ACh) for frontoparietal cortical networks thought to support attention. We examined a potential supramodal role of ACh in a frontoparietal cortical attentional network supporting target detection. We recorded local field potentials (LFPs) in the prelimbic frontal cortex (PFC) and the posterior parietal cortex (PPC) to assess whether ACh contributed to a state of readiness to alert rats to an impending presentation of visual or olfactory targets in one of five locations. Twenty male Long–Evans rats underwent training and then lesions of the NBM using the selective cholinergic immunotoxin 192 IgG-saporin (0.3 μg/μl; ACh-NBM-lesion) to reduce cholinergic afferentation of the cortical mantle. Postsurgery, ACh-NBM-lesioned rats had less correct responses and more omissions than sham-lesioned rats, which changed parametrically as we increased the attentional demands of the task with decreased target duration. This parametric deficit was found equally for both sensory targets. Accurate detection of visual and olfactory targets was associated specifically with increased LFP coherence, in the beta range, between the PFC and PPC, and with increased beta power in the PPC before the target's appearance in sham-lesioned rats. Readiness-associated changes in brain activity and visual and olfactory target detection were attenuated in the ACh-NBM-lesioned group. Accordingly, ACh may support supramodal attention via modulating activity in a frontoparietal cortical network, orchestrating a state of readiness to enhance target detection.SIGNIFICANCE STATEMENTWe examined whether the neurochemical acetylcholine (ACh) contributes to a state of readiness for target detection, by engaging frontoparietal cortical attentional networks independent of modality. We show that ACh supported alerting attention to an impending presentation of either visual or olfactory targets. Using local field potentials, enhanced stimulus detection was associated with an anticipatory increase in power in the beta oscillation range before the target's appearance within the posterior parietal cortex (PPC) as well as increased synchrony, also in beta, between the prefrontal cortex and PPC. These readiness-associated changes in brain activity and behavior were attenuated in rats with reduced cortical ACh. Thus, ACh may act, in a supramodal manner, to prepare frontoparietal cortical attentional networks for target detection.

The ability to capture the most relevant information from everyday experiences without constantly learning unimportant details is vital to survival and mental health. While decreased activity of the medial prefrontal cortex (mPFC) is associated with failed or inflexible encoding of relevant events in the hippocampus, mechanisms used by the mPFC to discern behavioral relevance of events are not clear. To address this question, we chemogenetically activated excitatory neurons in the mPFC of male rats and examined its impact on local network activity and differential associative learning dependent on the hippocampus. Rats were exposed to two neutral stimuli in two environments whose contingency with an aversive stimulus changed systematically across days. Over 2 weeks of differential and reversal learning, theta band activity began to ramp up toward the expected onset of the aversive stimulus, and this ramping activity tracked the subsequent shift of the set (stimulus modality to environment) predictive of the aversive stimulus. With chemogenetic mPFC activation, the ramping activity emerged within a few sessions of differential learning, which paralleled faster learning and stronger correlations between the ramping activity and conditioned responses. Chemogenetic mPFC activity, however, did not affect the adjustment of ramping activity or behavior during reversal learning or set-shifting, suggesting that the faster learning was not because of a general enhancement of attention, sensory, or motor processing. Thus, the dynamics of the mPFC network activation during events provide a relevance-signaling mechanism through which the mPFC exerts executive control over the encoding of those events in the hippocampus.

Background Near-infrared spectroscopy (NIRS) brain–computer interfaces (BCIs) enable users to interact with their environment using only cognitive activities. This paper presents the results of a comparison of four methodological frameworks used to select a pair of tasks to control a binary NIRS-BCI; specifically, three novel personalized task paradigms and the state-of-the-art prescribed task framework were explored. New methods Three types of personalized task selection approaches were compared, including: user-selected mental tasks using weighted slope scores (WS-scores), user-selected mental tasks using pair-wise accuracy rankings (PWAR), and researcher-selected mental tasks using PWAR. These paradigms, along with the state-of-the-art prescribed mental task framework, where mental tasks are selected based on the most commonly used tasks in literature, were tested by ten able-bodied participants who took part in five NIRS-BCI sessions. Results The frameworks were compared in terms of their accuracy, perceived ease-of-use, computational time, user preference, and length of training. Most notably, researcher-selected personalized tasks resulted in significantly higher accuracies, while user-selected personalized tasks resulted in significantly higher perceived ease-of-use. It was also concluded that PWAR minimized the amount of data that needed to be collected; while, WS-scores maximized user satisfaction and minimized computational time. Comparison with existing method In comparison to the state-of-the-art prescribed mental tasks, our findings show that overall, personalized tasks appear to be superior to prescribed tasks with respect to accuracy and perceived ease-of-use. Conclusions The deployment of personalized rather than prescribed mental tasks ought to be considered and further investigated in future NIRS-BCI studies.

As a noninvasive and safe optical measure of hemodynamic brain activity, near-infrared spectroscopy (NIRS) has emerged as a potential brain-computer interface (BCI) access modality. Currently, to the best of our knowledge, all NIRS BCIs use mental tasks to elicit changes in regional hemodynamic activity. One of the limitations of using mental tasks is that they can be cognitively demanding, and unintuitive. The goal of this work was to explore the development of a neurofeedback-based NIRS BCI that weans users off mental tasks, to instead use voluntary self-regulation. Ten able-bodied participants were recruited for this study. After ten sessions of using two personalized mental tasks to increase and decrease the participant’s hemodynamic activity, the users were asked, for the remaining sessions, to stop performing their tasks and instead use only a desire to modulate their hemodynamic activity. By the final online session, participants were able to exclusively use voluntary self-regulation with an average accuracy of $79 \pm 13\hbox{\%}$ . Additionally, the majority of participants indicated that BCI control via self-regulation was less taxing and more intuitive than BCI operation using mental tasks.

Deep brain stimulation (DBS) of the subgenual cingulate gyrus (SCG) has been used to treat patients with treatment-resistant depression. As in humans, DBS applied to the ventromedial prefrontal cortex of rats induces antidepressant-like responses. Physiological interactions between structures that play a role in depression and antidepressant treatment are still unknown. The present study examined the effect of DBS on inter-region communication by measuring the coherence of local field potentials in the rat infralimbic cortex (IL; homologue of the SCG) and one of its major afferents, the ventral hippocampus (VH). Rats received daily IL DBS treatment (100 μA, 90 μs, 130 Hz; 8 h/day). Recordings were conducted in unrestrained, behaving animals on the day before treatment, after 1 and 10 days of treatment, and 10 days stimulation offset. VH-IL coherence in the 2–4 Hz range was reduced in DBS-treated animals compared with shams after 10 days, but not after only 1 day of treatment. No effect of DBS was observed in the 6–10 Hz (theta) range, where coherence was generally high and could be further evoked with a loud auditory stimulus. Finally, coherence was not affected by fluoxetine (10 mg/kg), suggesting that the effects of DBS were not likely mediated by increased serotonin levels. While these data support the hypothesis that DBS disrupts communication between regions important for expectation-based control of emotion, they also suggest that lasting physiological effects require many days of treatment and, furthermore, may be specific to lower-frequency patterns, the nature and scope of which await further investigation.

Anatomical and electrophysiological studies collectively suggest that the entorhinal cortex consists of several subregions, each of which is involved in the processing of different types of information. Consistent with this idea, we previously reported that the dorsolateral portion of the entorhinal cortex (DLE), but not the caudomedial portion, is necessary for the expression of a memory association between temporally discontiguous stimuli in trace eyeblink conditioning (Morrissey et al. (2012) J Neurosci 32:5356-5361). The present study examined whether memory acquisition depends on the DLE and what types of local neurotransmitter mechanisms are involved in memory acquisition and expression. Male Long-Evans rats experienced trace eyeblink conditioning, in which an auditory conditioned stimulus (CS) was paired with a mildly aversive electric shock to the eyelid (US) with a stimulus-free interval of 500 ms. Immediately before the conditioning, the rats received a microinfusion of neuroreactive substances into the DLE. We found that reversible inactivation of the DLE with GABAA receptor agonist, muscimol impaired memory acquisition. Furthermore, blockade of local muscarinic acetylcholine receptors (mACh) with scopolamine retarded memory acquisition while blockade of local NMDA receptors with APV had no effect. Memory expression was not impaired by either type of receptor blocker. These results suggest that the DLE is necessary for memory acquisition, and that acquisition depends on the integrity of local mACh receptor-dependent firing modulation, but not NMDA receptor-dependent synaptic plasticity.

Following learning, increased coupling between spindle oscillations in the medial prefrontal cortex (mPFC) and ripple oscillations in the hippocampus is thought to underlie memory consolidation. However, whether learning-induced increases in ripple-spindle coupling are necessary for successful memory consolidation has not been tested directly. In order to decouple ripple-spindle oscillations, here we chemogenetically inhibited parvalbumin-positive (PV+) interneurons, since their activity is important for regulating the timing of spiking activity during oscillations. We found that contextual fear conditioning increased ripple-spindle coupling in mice. However, inhibition of PV+ cells in either CA1 or mPFC eliminated this learning-induced increase in ripple-spindle coupling without affecting ripple or spindle incidence. Consistent with the hypothesized importance of ripple-spindle coupling in memory consolidation, post-training inhibition of PV+ cells disrupted contextual fear memory consolidation. These results indicate that successful memory consolidation requires coherent hippocampal-neocortical communication mediated by PV+ cells., eLife digest Sleep contributes to the strengthening of memories. During non-dreaming sleep, neurons in regions of the brain that form and store memories – such as the hippocampus and prefrontal cortex – fire in rhythmic waves. The neurons in the hippocampus tend to fire during a wave that repeats up to 250 times per second, called sharp-wave ripples. Meanwhile, in the prefrontal cortex, the neurons tend to fire during a lower frequency wave that repeats 12 to 15 times per second, called spindles. During sleep and quiet wakefulness, hippocampal ripples often synchronize with prefrontal spindles; that is, both waves tend to occur at approximately the same time. Many neuroscientists think this allows the brain regions to better communicate with one another, which in turn should help the brain to strengthen memories. Consistent with this possibility, rodents that learn a new task show more synchrony between ripples and spindles afterwards. But no one had actually tested whether this increase in ripple-spindle synchrony does strengthen the rodent’s memory of the task. It was also unclear how the brain achieves such an increase. Xia et al. suspected that this process involved a group of inhibitory brain cells called parvalbumin-positive interneurons. These cells act like timekeepers, and help to synchronize the firing of groups of neurons. Xia et al. now show that training mice to associate an environment with a mild electric shock made it more likely that the animals would show ripple-spindle synchrony. Yet, inhibiting the activity of parvalbumin-positive interneurons in either the hippocampus or prefrontal cortex blocked this effect. It also prevented sleep from strengthening the animals’ memory of the link between the environment and the shock. Patients with Alzheimer’s disease have fewer parvalbumin-positive interneurons. By showing that these neurons help strengthen new memories, these findings may explain why losing them can impair memory. Restoring or replacing interneuron activity could be a promising therapeutic avenue to explore.

The lateral entorhinal cortex (LEC) is thought to bind sensory events with the environment where they took place. To compare the relative influence of transient events and temporally stable environmental stimuli on the firing of LEC cells, we recorded neuron spiking patterns in the region during blocks of a trace eyeblink conditioning paradigm performed in two environments and with different conditioning stimuli. Firing rates of some neurons were phasically selective for conditioned stimuli in a way that depended on which room the rat was in; nearly all neurons were tonically selective for environments in a way that depended on which stimuli had been presented in those environments. As rats moved from one environment to another, tonic neuron ensemble activity exhibited prospective information about the conditioned stimulus associated with the environment. Thus, the LEC formed phasic and tonic codes for event-environment associations, thereby accurately differentiating multiple experiences with overlapping features. DOI: http://dx.doi.org/10.7554/eLife.28611.001, eLife digest The context in which an event occurs plays a large role in how the brain understands and responds to the event. While a key part of context is where we are, contexts can also change within the same space: different meetings are held at different times and with different people in the same room, and a grassy field can be a place of intense competition or a place to relax and gaze at clouds. However, we have little understanding of how the brain sets up and maintains a sense of context. A region of the brain called the lateral entorhinal cortex (LEC) responds to events as they happen, but may also maintain a record of past experiences, and helps us to learn new associations between events. To find out how LEC neurons might represent context, Pilkiw et al. measured the activity of individual LEC neurons in rats as they experienced different combinations of events and environments. In each trial, the rats were placed in one of two different rooms and exposed to one of two sensory cues (sound or light) six times, either alone or, to test learning, paired moments later with a mild stimulation to the eyelid. The gaps between the cues lasted from 20 to 40 seconds. As expected, some LEC neurons responded to the sensory cues, and varied their responses to cues depending on whether or not they were paired with eyelid stimulation. What was much more striking is that almost all cells in the LEC behaved very differently in different contexts, both in response to the cues and also during the long gaps between the cues. This suggests that the LEC provides the brain with information about the circumstances of an event, and may be the reason we expect different events under different circumstances – even if we are in the same place. We tend to underestimate how much we rely on context to remember events and to guide our behavior. Many disabling health conditions, including addiction, post-traumatic stress disorder and obsessive-compulsive disorder, are affected by context. For example, people who are trying to overcome drug addiction can often reduce their cravings by avoiding places and situations in which they previously used the drug in question. Understanding how the LEC represents context may therefore help us to develop treatments that target this brain region in order to alter harmful behaviors. DOI: http://dx.doi.org/10.7554/eLife.28611.002

The entorhinal cortex is thought to support rapid encoding of new associations by serving as an interface between the hippocampus and neocortical regions. Although the entorhinal-hippocampal interaction is undoubtedly essential for initial memory acquisition, the entorhinal cortex contributes to memory retrieval even after the hippocampus is no longer necessary. This suggests that during memory consolidation additional synaptic reinforcement may take place within the cortical network, which may change the connectivity of entorhinal cortex with cortical regions other than the hippocampus. Here, I outline behavioral and physiological findings which collectively suggest that memory consolidation involves the gradual strengthening of connection between the entorhinal cortex and the medial prefrontal/anterior cingulate cortex (mPFC/ACC), a region that may permanently store the learned association. This newly formed connection allows for close interaction between the entorhinal cortex and the mPFC/ACC, through which the mPFC/ACC gains access to neocortical regions that store the content of memory. Thus, the entorhinal cortex may serve as a gatekeeper of cortical memory network by selectively interacting either with the hippocampus or mPFC/ACC depending on the age of memory. This model provides a new framework for a modification of cortical memory network during systems consolidation, thereby adding a fresh dimension to future studies on its biological mechanism.

Daily experiences are represented by networks of neurons distributed across the neocortex, bound together for rapid storage and later retrieval by the hippocampus. While the hippocampus is necessary for retrieving recent episode-based memory associations, over time, consolidation processes take place that enable many of these associations to be expressed independent of the hippocampus. It is generally thought that mechanisms of consolidation involve synaptic weight changes between cortical regions; or, in other words, the formation of "horizontal" cortico-cortical connections. Here, we review anatomical, behavioral, and physiological data which suggest that the connections in and between the entorhinal and cingulate cortices may be uniquely important for the long-term storage of memories that initially depend on the hippocampus. We propose that current theories of consolidation that divide memory into dual systems of hippocampus and neocortex might be improved by introducing a third, middle layer of entorhinal and cingulate allocortex, the synaptic weights within which are necessary and potentially sufficient for maintaining initially hippocampus-dependent associations over long time periods. This hypothesis makes a number of still untested predictions, and future experiments designed to address these will help to fill gaps in the current understanding of the cortical structure of consolidated memory.

Trace eyeblink conditioning requires animals to associate a neutral stimulus (CS) and an aversive periorbital shock (US) that occurred moment later. Acquisition in this conditioning depends on several forebrain regions including the hippocampus, medial prefrontal, and entorhinal cortices in addition to the cerebellum. Activities of single and population neurons in these regions show several patterns of change with the conditioning. For example, the power and synchronization of theta oscillations are correlated with the rate of acquisition. Yet, little is known about how neuronal oscillations at other frequency bands change with the conditioning. The present study examined changes in gamma oscillations, which are typically associated with spiking activity of individual cells. We found that after CS offset the power of gamma oscillations at 35-45 Hz fluctuated at about 7 Hz. This rhythmic fluctuation of gamma power was observed in all three regions and locked to local theta oscillations at 4-8 Hz. Furthermore, over the course of 10 days of acquisition sessions, the coupling of gamma power and theta phase became stronger in the medial prefrontal cortex while it did not change in the hippocampus or lateral entorhinal cortex. Neither theta nor gamma power in any of the regions significantly changed across sessions, rejecting a possibility that the observed increase in prefrontal theta-gamma coupling was secondary to an increase in theta or gamma power. The theta-gamma coupling between different regions did not significantly change across sessions. These results suggest that prefrontal gamma oscillations become more effectively coordinated with concurrent theta oscillations in trace eyeblink conditioning. This may result in a stronger impact of prefrontal neuronal firing responses to the CS on the processing in down-stream regions, such as the lateral entorhinal cortex and/or pontine nuclei.

This chapter reviews the past research toward identifying the brain circuit and its computation underlying the associative memory in eyeblink classical conditioning. In the standard delay eyeblink conditioning paradigm, the conditioned stimulus (CS) and eyeblink-eliciting unconditioned stimulus (US) converge in the cerebellar cortex and interpositus nucleus (IPN) through the pontine nuclei and inferior olivary nucleus. Repeated pairings of CS and US modify synaptic weights in the cerebellar cortex and IPN, enabling IPN neurons to activate the red nucleus and generate the conditioned response (CR). In a variant of the standard paradigm, trace eyeblink conditioning, the CS and US are separated by a brief stimulus-free trace interval. Acquisition in trace eyeblink conditioning depends on several forebrain regions, including the hippocampus and medial prefrontal cortex as well as the cerebellar–brainstem circuit. Details of computations taking place in these regions remain unclear; however, recent evidence supports a view that the forebrain encodes a temporal sequence of the CS, trace interval, and US in a specific environmental context and signals the cerebellar–brainstem circuit to execute the CR when the US is likely to occur. Together, delay eyeblink conditioning represents one of the most successful cases of understanding the neural substrates of long-term memory in mammals, while trace eyeblink conditioning demonstrates its utility for uncovering detailed computations in the whole brain network underlying long-term memory.

Previous work suggests that activation patterns of neurons in superficial layers of the neocortex are more sensitive to spatial context than activation patterns in deep cortical layers. A possible source of this laminar difference is the distribution of contextual information to the superficial cortical layers carried by hippocampal efferents that travel through the entorhinal cortex and subiculum. To evaluate the role that the hippocampus plays in determining context sensitivity in superficial cortical layers, behavior-induced expression of the immediate early gene Arc was examined in hippocampus-lesioned and control rats after exposing them to 2 distinct contexts. Contrary to expectations, hippocampal lesions had no observable effect on Arc expression in any neocortical layer relative to controls. Furthermore, another group of intact animals was exposed to the same environment twice, to determine the reliability of Arc-expression patterns across identical contextual and behavioral episodes. Although this condition included no difference in external input between 2 epochs, the significant layer differences in Arc expression still remained. Thus, laminar differences in activation or plasticity patterns are not likely to arise from hippocampal sources or differences in external inputs, but are more likely to be an intrinsic property of the neocortex.

Anatomical connectivity and single neuron coding suggest a segregation of information representation within lateral (LEC) and medial (MEC) portions of the entorhinal cortex, a brain region serving as the primary input/output of the hippocampus and maintaining widespread connections to many association cortices. The present study aimed to expand this idea by examining whether these two subregions differentially contribute to memory retrieval for an association between temporally discontiguous stimuli. We found that reversible inactivation of the LEC, but not the MEC, severely impaired the retrieval of the recently and remotely acquired memory in rat trace eyeblink conditioning, in which a stimulus-free interval was interposed between the conditioned and unconditioned stimulus. Conversely, inactivation of the LEC had no effect on retrieval in delay eyeblink conditioning, where two stimuli were presented without an interval. Therefore, the LEC, but not the MEC, plays a long-lasting role in the retrieval of a memory for an association between temporally discontiguous stimuli.

Anatomical and electrophysiological studies collectively suggest that the entorhinal cortex consists of several subregions, each of which is involved in the processing of different types of information. Consistent with this idea, we previously reported that the dorsolateral portion of the entorhinal cortex (DLE), but not the caudomedial portion, is necessary for the expression of a memory association between temporally discontiguous stimuli in trace eyeblink conditioning (Morrissey et al. (2012) J Neurosci 32:5356-5361). The present study examined whether memory acquisition depends on the DLE and what types of local neurotransmitter mechanisms are involved in memory acquisition and expression. Male Long-Evans rats experienced trace eyeblink conditioning, in which an auditory conditioned stimulus (CS) was paired with a mildly aversive electric shock to the eyelid (US) with a stimulus-free interval of 500 ms. Immediately before the conditioning, the rats received a microinfusion of neuroreactive substances into the DLE. We found that reversible inactivation of the DLE with GABAA receptor agonist, muscimol impaired memory acquisition. Furthermore, blockade of local muscarinic acetylcholine receptors (mACh) with scopolamine retarded memory acquisition while blockade of local NMDA receptors with APV had no effect. Memory expression was not impaired by either type of receptor blocker. These results suggest that the DLE is necessary for memory acquisition, and that acquisition depends on the integrity of local mACh receptor-dependent firing modulation, but not NMDA receptor-dependent synaptic plasticity.

Brain-computer interfaces (BCIs) allow individuals to use only cognitive activities to interact with their environment. The widespread use of BCIs is limited, due in part to their lack of user-friendliness. The main goal of this work was to develop a more user-centered BCI and determine if: (1) individuals can acquire control of an online near-infrared spectroscopy BCI via usability and performance-informed selection of mental tasks without compromising classification accuracy and (2) the combination of usability and performance-informed selection of mental tasks yields subjective ease-of-use ratings that exceed those attainable with prescribed mental tasks. Twenty able-bodied participants were recruited. Half of the participants served as a control group, using the state-of-the-art prescribed mental strategies. The other half of the participants comprised the study group, choosing their own personalized mental strategies out of eleven possible tasks. It was con- cluded that users were, in fact, able to acquire control of the more user-centered BCI without a significant change in accuracy compared to the prescribed task BCI. Furthermore, the personalized BCI yielded higher subjective ease-of-use ratings than the prescribed BCI. Average online accuracies of 77 � 12.9% and 73 � 12.9% were achieved by the personalized and prescribed mental task groups, respectively. © 2015 Society of Photo-Optical

The importance of the hippocampus in declarative memory is limited to recently acquired memory, and remotely acquired memory is believed to be stored somewhere in the neocortex. However, it remains unknown how the memory network is reorganized from a hippocampus-dependent form into a neocortex-dependent one. We reported previously that the medial prefrontal cortex (mPFC) is important for this neocortex-dependent remote memory in rat trace eyeblink conditioning. Here, we investigate the involvement of NMDA receptors in the mPFC in this reorganization and determine the time window of their contribution using chronic infusion of an antagonist into the mPFC, specifically during the postlearning consolidation period. The rats with blockade of the mPFC NMDA receptors during the first 1 or 2 weeks after learning showed a marked impairment in memory retention measured 6 weeks after learning, but relearned normally with subsequent conditioning. In contrast, the same treatment had no effect if it was performed during the third to fourth weeks or during the first day just after learning. The specificity of NMDA receptor blockade was confirmed by the reduced long-term potentiation in the hippocampal-prefrontal pathway in these rats. These results suggest that successful establishment of remotely acquired memory requires activation of NMDA receptors in the mPFC during at least the initial week of the postlearning period. Such NMDA receptor-dependent processes may mediate the maturation of neocortical networks that underlies permanent memory storage and serve as a way to reorganize memory circuitry to the neocortex-dependent form.

The prefrontal cortex is implicated in many cognitive functions, including working memory, the temporal organization of behavior, and emotion (Kolb 1984; Goldman-Rakic 1990; Fuster 1997), and also is presumed to have the most dynamic and richest interaction with other brain regions (Fuster 2001). Recent reports have expanded the importance of the rabbit and rodent medial prefrontal cortex (mPFC) to hippocampus-dependent motor learning, e.g., trace classical eyeblink conditioning. Since the essential neuronal circuitry for eyeblink conditioning is relatively well characterized, investigating the role of the mPFC in this task may contribute to our understanding of the nature of the mPFC's interaction with other brain areas. Trace eyeblink conditioning is a hippocampus-dependent variation of classical eyeblink conditioning, a process that depends critically on the cerebellum and brainstem (for review, see Thompson et al. 1997). In this paradigm, the conditioned stimulus (CS) and the unconditioned stimulus (US) are separated by a stimulus-free trace interval. Previous studies using permanent lesion methods have revealed that pre-conditioning lesioning of the hippocampus (Solomon et al. 1986; Moyer et al. 1990; McGlinchey-Berroth et al. 1997; Beylin et al. 2001), the medial prefrontal cortex (mPFC) (Kronforst-Collins and Disterhoft 1998; Weible et al. 2000, 2003; McLaughlin et al. 2002), the entorhinal cortex (Ryou et al. 2001), and the mediodorsal thalamus (Powell and Churchwell 2002) all impair acquisition of the trace eyeblink-conditioned response (CR), suggesting that the circuitry for this learning covers multiple regions of the brain, as others have previously proposed (Weiss and Disterhoft 1996; Green and Woodruff-Pak 2000). Furthermore, this circuitry is reorganized after the CR has been completely acquired. Retention of a recently acquired CR is mediated mainly by the hippocampus (Kim et al. 1995; Takehara et al. 2002, 2003a) and the cerebellum (Woodruff-Pak et al. 1985; Takehara et al. 2003a), but only marginally by the mPFC (Powell et al. 2001; Takehara et al. 2003a). Retention of a remotely acquired CR is mediated by the mPFC and the cerebellum (Takehara et al. 2003a). Although these previous studies using permanent lesion methods have indicated an involvement of the mPFC in trace eyeblink conditioning, this method has only limited utility when we seek a detailed understanding of the participation of the mPFC, because permanent lesions affect not only acquisition, but also consolidation and retrieval. Therefore, further studies that can specify the stage of memory and also elucidate the underlying molecular process are required for a more detailed understanding of the mPFC's precise role. Here we investigated the involvement of the mPFC in mnemonic processes during and after daily conditioning using microinfusion of the γ-aminobutyric acid type A (GABAA) receptor agonist muscimol. In addition, we examined the involvement of N-Methyl-d-aspartate (NMDA) receptors in this process using microinfusion of the NMDA receptor antagonist 2-amino-5-phosphonovaleric acid (APV), because synaptic plasticity in the prelimbic area of the mPFC depends on NMDA receptors (Hirsch and Crepel 1991; Jay et al. 1995). A preliminary report of some of these data has been presented previously in abstract form (Takehara et al. 2003b).