Neurobiology of Learning and Memory

Congruency of newly learned information with previous knowledge (i.e. a mental schema) leads to facilitated encoding and rapid integration into neocortical memory networks. It is less known whether this is associated with a differential involvement of the hippocampus in consolidation of schema-congruent and -incongruent information. Here, we used the GABAA-ergic anesthetic propofol to transiently modulate hippocampal neural activity shortly after encoding of schema-congruent and -incongruent information in human patients. We found a significant difference in memory of schema-congruent and -incongruent words in patients that was absent in controls. This effect was driven by a benefit for schema-congruent words, thus suggesting that propofol administration facilitated consolidation of previously encoded schema-congruent items. Our results suggest that schema-congruency of newly learned information significantly modulates involvement of hippocampus-dependent networks during memory consolidation. They further support the hypothesis of a competitive interaction between hippocampus and extra-hippocampal networks during early memory consolidation.

This study examined the effect of danger on consolidation of neutral information in two regions of the rat (male and female) medial temporal lobe: the perirhinal cortex (PRh) and basolateral amygdala complex (BLA). The neutral information was the association that forms between an auditory stimulus and a visual stimulus (labelled S2 and S1) across their pairings in sensory preconditioning. We show that, when the sensory preconditioning session is followed by a shocked context exposure, the danger shifts consolidation of the S2-S1 association from the PRh to the BLA; and does so by interacting with processes involved in encoding of the S2-S1 pairings. Specifically, we show that the initial S2-S1 pairing in sensory preconditioning is encoded in the BLA and not the PRh; whereas the later S2-S1 pairings are encoded in the PRh and not the BLA. When the sensory preconditioning session is followed by a context alone exposure, the BLA-dependent trace of the early S2-S1 pairings decays and the PRh-dependent trace of the later S2-S1 pairings is consolidated in memory. However, when the sensory preconditioning session is followed by a shocked context exposure, the PRh-dependent trace of the later S2-S1 pairings is suppressed and the BLA-dependent trace of the initial S2-S1 pairing is consolidated in memory. These findings are discussed with respect to mutually inhibitory interactions between the PRh and BLA, and the way that these regions support memory in other protocols, including recognition memory in people. Significance StatementThe perirhinal cortex (PRh) and basolateral amygdala complex (BLA) process the pairings of neutral auditory and visual stimuli in sensory preconditioning. The involvement of each region in this processing is determined by the novelty/familiarity of the stimuli as well as events that occur immediately after the preconditioning session. Novel stimuli are represented in the BLA; however, as these stimuli are repeatedly presented without consequence, they come to be represented in the PRh. Whether the BLA- or PRh-dependent representation is consolidated in memory depends on what happens next. When nothing of significance occurs, the PRh-dependent representation is consolidated and the BLA-dependent representation decays; but when danger is encountered, the PRh-dependent representation is inhibited and the BLA-dependent representation is selected for consolidation.

The loss of the dorsal hippocampus (DH) results in profound deficits in contextual fear-conditioned (CxFC) memory. Nonetheless, CxFC memories can still form without the DH, specifically with multiple trials at three-day intervals. The infralimbic cortex (IL) is pivotal in initiating this compensatory process post-DH loss, but the precise factors remain elusive. Our study aims to delineate key factors of compensatory CxFC in DH absence by investigating the effects of sleep deprivation (SD) and NMDA receptor subunits NR2A and NR2B. Using a DH-lesioned rat model, we conducted two conditioning trials separated by three days and assessed fear response during the subsequent test. We observed that DH-lesioned animals exhibited to SD (DHL-SD) did not elicit a compensatory CxFC response, displaying significantly impaired freezing during the second test. Conversely, DH-lesioned non-sleep-deprived animals (DHL-NSD) compensated for DH loss and exhibited robust CxFC responses during the second test. Moreover, inhibiting NR2B subunits in the IL during initial CxFC training disrupted the formation of compensatory fear memory in DH-lesioned animals, while NR2A subunit blockade showed no significant effect. These emphasize the adverse impact of SD on compensatory memory and the critical role of NR2B subunits in facilitating compensatory CxFC memory formation following DH loss.

Formation and consolidation of memories for highly stressful (traumatic) events is a complex process that involves interplay between multiple memory systems and has implications for etiology and treatment of stress- and trauma-related disorders. Here we study effects of sleep/wake state and high intra-hippocampal corticosterone on consolidation of aversive contextual memories as well as consolidation of association between simple trauma-related cues and fear response in rats. Animals were implanted with EEG and EMG electrodes for sleep assessment and cannulas for intra-hippocampal corticosterone application. They were familiarized to a "safe box" and then trained in fear conditioning paradigm in a distinct "shock box" with a prominent simple auditory cue serving as a phasic background cue. Immediately after conditioning, animals received bilateral intra-hippocampal saline (1l) or corticosterone (10ng in 1l saline) injection and were either allowed to sleep or were kept awake for a following two-hour consolidation period. Memory test twenty-four hours later revealed that the saline-injected animals with sleep during consolidation had significantly stronger freezing response in the shock box compared to the safe box as well as increased freezing in response to the tone. Lack of post-learning sleep in saline injected animals led to generalization of fear response to the safe context, while association between simple cue and fear response was preserved. High intra-hippocampal corticosterone level during memory consolidation led to generalization of fear response to the safe context, regardless of sleep/wake state, while enhancement of response to single stimulus was not observed. Our results show how manipulation of conditions during consolidation can lead to greatly variable complex memories for a traumatic episode and distinct behavioral outcomes. HighlightsO_LIWe studied effect of sleep and intrahippocampal corticosterone on consolidation of memories surrounding stressful event modeled by fear conditioning in rats. C_LIO_LISleep following traumatic fear conditioning event is important for subsequent manifestation of fear response (freezing) specifically in the context of traumatic event but not in a neutral safe context. C_LIO_LILack of sleep or high intra-hippocampal corticosterone level during memory consolidation leads to generalization of fear response to both the traumatic and safe context. C_LIO_LIIncreased freezing in response to a trauma-related auditory cue was observed in saline injected rats regardless of wake/sleep state during consolidation. C_LIO_LIPost-learning intra-hippocampal corticosterone injection blocked response to a trauma-related auditory cue regardless of wake/sleep state during consolidation. C_LI

The perirhinal and postrhinal cortices reside in the parahippocampal region of the medial temporal lobe. They receive and process sensory information from cortical and subcortical sources and support hippocampal functions via direct connectivity and indirectly via the entorhinal cortex. Previous studies using the sensory preconditioning paradigm have shown that the perirhinal and postrhinal cortices are necessary for associating cues during preconditioning and retrieving the associations during conditioning. However, the question of whether these regions are also required for stimulus association retrieval during re-exposure to the preconditioning cue had not been addressed. Using a chemogenetic approach, we temporarily suppressed the perirhinal or postrhinal cortex in adult male rats during the preconditioning cue test phase. Both suppression groups showed impaired sensory preconditioning compared to sham surgery controls, as indicated by significantly reduced preferential responding. Implications of our findings are discussed in relation to the importance of the PER and POR in context processing and episodic-like memory in animal models. Significance statementAlthough Pavlovian conditioning between two stimuli is widely used in neuroscience, associative learning in the real-world is often too complex to be modeled by first-order conditioning alone. Proper responding may require integrating multiple associations via common elements, i.e. higher-order conditioning, the neural basis of which is not well understood. Whereas existing research indicates the perirhinal and postrhinal cortices in the medial temporal lobe contribute to certain forms of sensory preconditioning (SPC, a type of higher-order conditioning), we suggest their contributions lie in the encoding and retrieving latently acquired associations, processes which are central to all forms of SPC. We argue this interpretation also captures perirhinal and postrhinal contributions to contextual and episodic memory, hence offering a unifying explanation regarding the contributions of these cortices across multiple paradigms.

Across sleep-based and retrieval-mediated consolidation, memories typically become generalised and less dependent on their episodic components for their recollection. However, memory transformations across sleep and retrieval training have not been directly compared. The current study aims to compare how sleep and retrieval training impact the endorsement of semantically similar and different lures, as well as their episodic recollection using the parietal old/new effect on the late positive component (LPC) in subjects EEG. Thirty subjects (27F, 18-34, M=22.17) attended four sessions where they learnt different sets of 104 object-word pairs and completed one of four 120-minute memory interventions: retrieval training (i.e., cued recall practice), restudy (i.e., pair re-exposure), a nap opportunity, or a wakeful rest. EEG was recorded while subjects were tested on their recognition accuracy in an old/new paradigm with similar- and different-object lures. Our results revealed that retrieval training, but not sleep, lead to greater accuracy for identifying old pairs, but worse similar-lure discrimination. Whilst the parietal old/new effect did not differ between conditions, retrieval had lower LPC amplitudes for similar- than different-object false alarms, whilst restudy demonstrated the opposite. Sleep and wake demonstrated no differences in LPC amplitudes between hits and different false alarm types. Together, our study demonstrates evidence for gist-abstraction across retrieval training, and a task-relevant selective maintenance of episodic details across sleep. These results challenge theories that retrieval training replicates sleep-based consolidation mechanisms, instead acting as a fast route to semanticization regardless of the context.

Damage to the hippocampus produces profound retrograde amnesia, but odour and object discrimination memories can be spared in the retrograde direction. Prior lesion studies testing retrograde amnesia for object/odour discriminations are problematic due to sparing of large parts of the hippocampus, which may support memory recall, and/or the presence of uncontrolled, distinctive odours that may support object discrimination. To address these issues, we used a simple object discrimination test to assess memory in male rats. Two visually distinct objects, paired with distinct odour cues, were presented. One object was associated with a reward. Following training, neurotoxic hippocampal lesions were made using N-methyl-D-aspartate (NMDA). The rats were then tested on the preoperatively learned object discrimination problem, with and without the availability of odour or visual cues during testing. The rats were also postoperatively trained on a new object discrimination problem. Lesion sizes ranged from 67-97% of the hippocampus (average of 87%). On the preoperatively learned discrimination problem, the rats with hippocampal lesions showed preserved object discrimination memory when tested in the dark (i.e., without visual cues) but not when the explicit odour cues were removed from the objects. Hippocampal lesions increased the number of trials required to reach criterion but did not prevent rats from solving the postoperatively learned discrimination problem. Our results support the idea that long-term memories for odours, unlike recall of visual properties of objects, does not depend on the hippocampus in rats, consistent with previous observations that hippocampal damage does not cause retrograde amnesia for odour memories.

It is widely accepted that fear memories are consolidated through protein synthesis-dependent changes in the basolateral amygdala complex (BLA). However, recent studies show that protein synthesis is not required to consolidate the memory of a new dangerous experience when it is similar to a prior experience. Here, we examined whether the protein synthesis requirement for consolidation of the new experience varies with its spatial and temporal distance from the prior experience. In each experiment, rats were conditioned to fear a stimulus (S1, e.g., light) across its pairings with shock in stage 1; and a second stimulus (S2, e.g., tone) that preceded additional S1-shock pairings (S2-S1-shock) in stage 2. The latter stage was followed by a BLA infusion of a protein synthesis inhibitor, cycloheximide or vehicle. Finally, rats were tested for fear to S2. Critically, protein synthesis in the BLA was not required to consolidate fear to S2 when the two training stages occurred 48 hours apart and in the same context; was required when the two training stages were separated by a 14-day delay or occurred in different contexts; but was again not required when rats were re-exposed to S1 or shock after the delay or in the different context. Thus, protein synthesis in the BLA is not always required to consolidate a new fear memory. Instead, this requirement is determined by the degree of similarity between present and past experiences, the time and place in which those experiences occur, as well as reminders of the past experience. Significance StatementProtein synthesis in the basolateral amygdala complex (BLA) is not required to consolidate the memory of a new dangerous experience when it is similar to a prior experience. This study is significant in showing that: 1) when the new, similar experience occurs after a delay or in a different context, the protein synthesis requirement for its consolidation is reinstated; and 2) the effects of the delay and context shift are reversed by reminding animals of their prior experience. Thus, the neural mechanisms underlying memory consolidation are dynamically regulated by similarity/dissimilarity between present and past experiences, the time and place in which those experiences occur, as well as reminders of the past experience.

The partial reinforcement extinction effect (PREE) is a paradoxical learning phenomenon in which omission of reinforcement during acquisition results in more persistent conditioned responding in extinction. Here, we report a significant PREE with an inverted-U, entropy-like distribution against reinforcement probability following tone foot shock fear conditioning in rats, which was associated with increased neural activity in hippocampus and amygdala as indexed by p-ERK and c-fos immunolabelling. In vivo electrophysiological recordings of local field potentials (LFPs) showed that 50% reinforcement was associated with increases in the frequency and power of tone-evoked theta oscillations in both the subiculum region of hippocampus and in basolateral amygdala (BLA) during both acquisition (Day 1) and extinction (Day 2) sessions. Tone-evoked LFPs in 50% reinforced animals also showed increases in coherence and bidirectional Granger Causality between hippocampus and amygdala. The results support a Bayesian interpretation of the PREE, in which the phenomenon is driven by increases in the entropy or uncertainty of stimulus contingencies, and indicate a crucial role for hippocampus in mediating this uncertainty-dependent effect.

After encoding, memories go through a labile state followed by a stabilization process known as consolidation. Once consolidated they can enter a new labile state after the presentation of a reminder of the original memory, followed by a period of restabilization (reconsolidation). During these periods of lability the memory traces can be modified. Currently, there are studies that show a rapid stabilization after 30 min, while others show that stabilization occurs after longer periods (e.g. 6 h). Here we investigate the effect of an interference treatment on declarative memory consolidation, comparing distinct time intervals after acquisition. On day 1, participants learned a list of non-syllable pairs (List 1). Immediately after, 30 min, 3 h or 8 h later, they received an interference list (List 2) that acted as an amnesic agent. On day 2 (48 h after training) participants had to recall List 1 first, followed by List 2. We found that the List 1 memory was susceptible to interference when the List 2 was administered immediately or 3 h after learning; however, shortly after acquisition (e.g. 30 min) the List 1 memory becomes transiently protected against interference. We propose the possibility that this rapid memory protection could be induced by a fast and transient neocortical integration (where the memory is transiently protected) becoming partially independent from the hippocampus followed by a hippocampal re-engagement where the memory becomes susceptible to interferences again. Our results open a discussion about the contribution of molecular and systemic aspects to memory consolidation.

Memory consolidation refers to the process by which newly encoded memories are strengthened and retained over time, and ample evidence indicates that sleep supports this process for both procedural and declarative memories. Although sleep spindles during non-rapid eye movement (NREM) sleep have been associated to consolidation, it remains unclear whether all spindle types contribute equally. Spindles vary in frequency and topography-slow spindles ([&le;]12.5Hz) predominating over frontal regions, whereas fast spindles (>12.5Hz) peak parietally - and recent work suggests that procedural memory consolidation during overnight sleep is related to the temporal organization of spindles in trains (i.e., events occurring <6s apart). Here we investigated whether a similar mechanism operates for declarative memory during daytime naps. Participants were assigned to a Nap (N=23) or No-Nap (N=15) group, and completed an object-spatial location task involving 36 item-location associations. Memory was assessed immediately after learning and again following a 90-minute nap or an equivalent wake period. Results showed that the Nap group exhibited significantly better delayed memory, as measured by combined recall-recognition score, and a greater proportion of participants maintained or improved their performance. In the Nap group, memory performance correlated with local spindle density at frontal and parietal sites, and, critically, with the proportion of slow spindles clustered in trains during NREM2. These findings suggest the temporal organization of slow spindles into clusters support declarative memory consolidation, pointing to a shared spindle-based mechanism across domains.

Aversive associative learning paradigms such as inhibitory avoidance (IA) are frequently used to examine episodic-like memories in rodents. In IA, rodents learn to associate a context with a footshock, followed by testing for memory strength in the original training context and for memory precision in a similar yet distinct neutral context. The present work assessed the effects of different contextual exposure procedures on memory strength and precision in IA at both recent and remote time points using male and female Long-Evans rats. An initial experiment found that rats kept in the lit (non-shock) compartment of the IA apparatus for 60 s during training, as opposed to 10 s, displayed enhanced memory strength, with discrimination between both chambers at the recent retention test and generalization at the remote retention test. Subsequent experiments investigated the effects of contextual pre-exposure the day before training. The results indicate that pre-exposure to the neutral context promoted generalization without altering memory strength compared to the first experiment. In contrast, pre-exposure to the aversive chamber promoted discrimination and enhanced memory strength. Notably, the different procedures yielded similar effects in both sexes. However, the results also indicate an overall pattern of greater contextual discrimination in females compared to males. These findings provide evidence for how different contextual exposures influence the degree of encoding at the time of training and a behavioral foundation for future studies examining the neurobiological mechanisms underlying memory strength and precision in IA, while highlighting the importance of using both sexes in initial behavioral work. Significance StatementStrength and precision are two fundamental properties of memory that can be simultaneously measured using inhibitory avoidance (IA), a type of context-footshock association task. However, little is known about how different context exposures alter rats encoding of these memories, thereby influencing subsequent memory strength and precision. Here, we found that pre-exposure to the neutral IA chamber decreased memory precision, whereas pre-exposure to the aversive IA chamber promoted memory strength and precision. Additionally, females demonstrated overall enhanced memory precision compared to males. These results indicate that different types of contextual exposures influence initial IA encoding and add to a limited body of research examining memory strength and precision in IA in both sexes.

Return of fear poses a problem for extinction-based therapies of clinical anxiety. Experimental research has discovered several pathways to return of fear, one of which is known as reinstatement. Here, we evaluated in rats the potential of scopolamine, a non-selective muscarinic receptor antagonist that is also safe for use in humans, to prevent the reinstatement of extinguished fear. We conducted three experiments with a total sample of 96 female rats. All rats went through a fear acquisition session (tone-shock pairings, CS-US), followed by two extinction sessions (CS only) and a post-extinction fear memory test. Twenty-four hours later, rats were placed in the same or a different context from extinction and received two unsignaled foot shock (US) presentations. On the following day, CS-evoked freezing returned when the reinstating USs had occurred in the same context compared to a different context (context-dependent reinstatement, Experiment 1). Systemic administration of scopolamine before or after the reinstating USs blocked the return of CS-evoked freezing on the following day (Experiments 2 and 3). Our findings suggest that administering scopolamine around the time of an aversive experience could prevent relapse of extinguished fears in humans.

This study examined effects of epinephrine injected in learning under anesthesia on awake memory of three fear conditioning tasks. Epinephrine given after training at various doses did not affect conditioned freezing acquired in an awake state or under 50 mg/kg pentobarbital; however, 0.1 mg/kg epinephrine caused saving in re-learning. For conditioned-fear potentiation of startle acquired in an awake state, epinephrine at 0.01 or 0.1 mg/kg enhanced 1-trial learning but at 1.0 mg/kg impaired 5-trial learning. For this task trained under pentobarbital, epinephrine enhanced 1-trial learning at 1.0 mg/kg and 5-trial learning at 0.1 or 1.0 mg/kg. Under infusion of dexmedetomidine (0.1 mg/kg/hr), learning of this task at a 0.63 mA foot shock yielded poor memory, which was improved by 0.1 mg/kg epinephrine; yet epinephrine had no effect on the better memory yielded by 1.25 mA foot shock. In an inhibitory avoidance task, rats in an awake state explored the apparatus and then received foot shocks under anesthesia of dexmedetomidine, injections of 0.1 mg/kg epinephrine before each shock training session enhanced memory. Deleting the awake exploration abolished the epinephrine effect. These results, taken together, suggest that epinephrine could enhance learning in three conditioning tasks under two anesthetics when the fear memory was assessed indirectly through its modification of an innate or learned behavioral tendency.

Extinction is a fundamental form of inhibitory learning that is important for adapting to changing environmental contingencies. While numerous studies have investigated the neural correlates of extinction using Pavlovian fear conditioning and appetitive operant reward-seeking procedures, less is known about the neural circuitry mediating the extinction of appetitive Pavlovian conditioned responding. Here, we aimed to generate an extensive brain activation map of extinction learning in a rat model of appetitive Pavlovian conditioning. Male Long-Evans rats were trained to associate a conditioned stimulus (CS; 20 s white noise) with the delivery of a 10% sucrose unconditioned stimulus (US; 0.3 ml/CS) to a fluid port. Control groups also received CS presentations, but sucrose was delivered either during the inter-trial interval or in the home-cage. After conditioning, 1 or 6 extinction sessions were conducted in which the CS was presented but sucrose was withheld. We performed Fos immunohistochemistry and network connectivity analyses on a set of cortical, striatal, thalamic, and amygdalar brain regions. Neural activity in the prelimbic cortex, ventral orbitofrontal cortex, nucleus accumbens core, and paraventricular nucleus of the thalamus was greater during recall relative to extinction. Conversely, prolonged extinction following 6 sessions induced increased neural activity in the infralimbic cortex, medial orbitofrontal cortex, and nucleus accumbens shell compared to home-cage controls. All these structures were similarly recruited during recall on the first extinction session. These findings provide novel evidence for the contribution of brain areas and neural networks that are differentially involved in the recall versus extinction of appetitive Pavlovian conditioned responding.

Human memory for recent events is believed to undergo reactivation during sleep. This process is thought to be relevant for the consolidation of both individual episodic memories and gist extraction, the formation of generalized memory representations from multiple, related memories. Which kinds of gist are actually enhanced, however, is the subject of less consensus. To address this question, we focused our design on four types of gist: inferential gist (relations extracted across non-contiguous events), statistical learning (regularities extracted from a series), summary gist (a theme abstracted from a temporally contiguous series of items), and category gist (characterization of a stimulus at a higher level in the semantic hierarchy). Sixty-nine participants (30 men, 38 women, and 1 other) completed memory encoding tasks addressing these types of gist and corresponding retrieval tasks the same evening, the morning after, and one week later. Inferential gist was retained over a week, whereas memory for category gist, summary gist, and statistical learning decayed. Higher proportions of REM were associated with worse performance in a statistical learning task controlling for time. Our results support that REM sleep is involved in schema disintegration, which works against participants ability to identify regularities within temporal series.\n\nSIGNIFICANCE STATEMENTTo gain the most from our experiences, we extract from them the most important elements, or \"gist\", with sleep believed to facilitate this process. However, what is referred to as gist varies considerably across studies. We report categorically different mnemonic trajectories of two classes of gist. In particular, we show that gist involving synthesis across relational memories is retained over time, whereas other gists were subject to substantial decay. Moreover, our evidence supports the idea that REM works to discretize, rather than synthesize experiences. Future research should test similar constructs in different tasks to determine whether these findings are generalizable. Our research suggests that patients with reduced REM sleep may experience more interference between similar memories.

Anxiety disorders (ADs) are among the most prevalent mental health conditions, yet first-line treatments often yield only moderate effectiveness. The fear conditioning paradigm is commonly used to investigate fear and extinction learning, revealing deficits in these processes and dysfunctional activity in the ventromedial prefrontal cortex (vmPFC) and amygdala in individuals with ADs. The vmPFC plays a critical role in regulating activity of the amygdala and consolidation of fear extinction memory. Notably, transcranial direct current stimulation (tDCS) has shown promise in enhancing fear extinction by modulating vmPFC activity. Additionally, rapid eye movement (REM) sleep has been suggested to be crucial for fear extinction memory consolidation. This study investigated the role the vmPFC during REM sleep in fear extinction memory consolidation. Thirty-two participants underwent a 3-day differential fear conditioning paradigm, with tDCS or sham stimulation applied during REM sleep over the vmPFC. Outcome measures included skin conductance responses (SCR) and subjective ratings of arousal, fear, and valence. Results indicate that tDCS during REM sleep enhances fear extinction memory consolidation, as measured by SCR. Furthermore, participants reported an increased subjective arousal following tDCS. These findings suggest that tDCS during REM sleep may hold potential for improving exposure-based treatments for ADs by strengthening fear extinction memory.

Adaptive behavior often necessitates that animals learn about events in a manner that is specific to a particular context or environment. These hierarchical organizations allow the animal to decide which action is the most appropriate when faced with ambiguous or conflicting possibilities. This study examined the role of hippocampus in enabling animals to use the context to guide action selection. We used a hierarchical instrumental outcome devaluation task in which male rats learn that the context provides information about the unique action-outcome relations that are in effect. We first confirmed that rats encode and use hierarchical context-(action-outcome) relations. We then show that chemogenetic inhibition of ventral hippocampus (vHPC) impairs both the encoding and retrieval of these associations, while inhibition of dorsal hippocampus (dHPC) impairs only the retrieval. Importantly, neither dHPC or vHPC were required for goal-directed behavior per se as these impairments only emerged when rats were forced to use the context to identify the current action-outcome relationships. These findings are discussed with respect to the role of the hippocampus and its broader circuitry in the contextual modulation of goal-directed behavior and the importance of hierarchical associations in flexible behavior.

Sleep, especially slow-wave sleep (SWS), has been found to facilitate memory consolidation for many types of learning. Mathematical learning, however, has seldom been examined in this context. Solving multiplication problems involves multiple steps before problems can be mastered or answers memorized, and thus it can depend on both skill learning and fact learning. Here we aimed to test the hypothesis that memory reactivation during sleep contributes to multiplication learning. To do so, we used a technique known as targeted memory reactivation (TMR), or the pairing of newly learned information with specific stimuli that are later presented during sleep. With TMR, specific memories can be reactivated over a period of sleep without disrupting ongoing sleep. We applied TMR during an afternoon nap to reactivate half of the multiplication problems that had previously been practiced. Results showed no effect of TMR on response time or accuracy of multiplication problem solving. Because these results were unexpected, we also used a variation of this paradigm to examine results in subjects who remained awake. Comparisons between the wake and sleep groups showed no difference in response time or accuracy in either the initial test or the final test. Although neither TMR nor sleep differentially influenced multiplication performance, correlational analysis provided some clues about mathematical problem solving and sleep. On the basis of these findings, even though they did not provide convincing support for our hypotheses, we suggest future experiments that could help produce a better understanding of the relevance of sleep and memory reactivation for this type of learning.

Associative recognition memory allows us to form representations of items and their environment and to judge the novelty of such representations. This memory is dependent on a brain circuit that includes interactions between medial prefrontal cortex (mPFC) and lateral entorhinal cortex (LEC); however it is unknown whether the interaction of these brain areas is required for memory encoding, retrieval or both processes. Furthermore, little is known as to whether indirect or direct mPFC-LEC connections are critical for associative recognition memory and, if the latter, in which direction information travels. To address these questions, we first performed pharmacological disconnection of mPFC and LEC, finding that mPFC-LEC interaction is required for both memory encoding and retrieval. Next, we optogenetically inhibited projections from mPFC to LEC, showing that this projection was crucial for both encoding and retrieval of both object-in-place and object-in-context recognition memory when a 1 h, but not a 5 min, memory retention delay was used. These data show that a direct connection from mPFC to LEC is critical for associative recognition memory, in a delay-dependent manner.