Validation of a rodent model of episodic memory replay.

Vivid episodic memories in humans have been described as the replay of the flow of past events in sequential order. Recently, Panoz-Brown et al. Current Biology, 28, 1628-1634, (2018) developed an olfactory memory task in which rats were presented with a list of trial-unique odors in an encoding context; next, in a distinctive memory assessment context, the rats were rewarded for choosing the second to last item from the list while avoiding other items from the list. In a different memory assessment context, the fourth to last item was rewarded. According to the episodic memory replay hypothesis, the rat remembers the list items and searches these items to find the item at the targeted locations in the list. However, events presented sequentially differ in memory trace strength, allowing a rat to use the relative familiarity of the memory traces, instead of episodic memory replay, to solve the task. Here, we directly manipulated memory trace strength by manipulating the odor intensity of target odors in both the list presentation and memory assessment. The rats relied on episodic memory replay to solve the memory assessment in conditions in which reliance on memory trace strength is ruled out. We conclude that rats are able to replay episodic memories.

Replay of incidentally encoded episodic memories in the rat.

Although events are not always known to be important when they occur, people can remember details about such incidentally encoded information using episodic memory. Importantly, when information is explicitly encoded for use in an expected test of retention (as in most assessments in animals), it is possible that it is used to generate a planned action1,2,3; thus, the remembered action can occur without remembering the earlier episode. By contrast, when a test is unexpected, transforming information into an action plan is unlikely because the importance of the information and the nature of the test are not yet known. Thus, accurate performance in an unexpected test after incidental encoding documents episodic memory.1,2,3,4,5,6,7,8 Here, we present evidence that rats replay episodic memories of incidentally encoded information in an unexpected assessment of memory. In one task,9 rats reported the third-last item in an explicitly encoded list of trial-unique odors. In a second task,10 rats foraged in a radial maze in the absence of odors. On a critical test, rats foraged in the radial maze, but scented lids covered the food. Next, memory of the third-last odor was assessed. All participating rats correctly answered the unexpected question. These results suggest that rats encoded multiple pieces of putatively unimportant information, and later they replayed a stream of episodic memories when that information was needed to solve an unexpected problem. We propose that rats replay episodic memories of incidentally encoded information, which documents a critical aspect of human episodic memory in a non-human animal.

Effects of NMDA antagonist dizocilpine (MK-801) are modulated by the number of distractor stimuli in the rodent odor span task of working memory.

The rodent odor span task (OST) uses an incrementing non-matching to sample procedure in which a series of odors is presented and selection of the session-novel odor is reinforced. An OST is frequently used to test the effects of neurobiological variables on memory capacity as the number of odors to remember increases during the course of the session. In this regard, one important finding has been that NMDA receptor antagonists selectively impair OST performance at doses that spare accuracy on control tasks. However, in many versions of the odor span task the number of stimuli to remember is confounded with the number of distractor odors presented to the rat on each trial. The present study compared the effects of the NMDA antagonist dizocilpine when the number of choices was held constant at two (one novel odor-S+ and one previously presented distractor odor-S-) and when the number of choice stimuli was permitted to increase up to 10 (one S+ and 9 S-). Dizocilpine impaired OST accuracy at doses that had no effect on a reference memory control task in both 2-choice and 10-choice conditions; however, the dose-response function was shifted to the left in the 10-choice tests. The impairments produced by dizocilpine were exacerbated as the memory load increased in both 2- and 10-choice conditions. These findings support the hypothesis that NMDA antagonism reduces the number of stimuli that rats can remember accurately, but the interaction between the effective DZP dose and the number of distractors shows that drug effects on OST performances may involve attentional factors in addition to memory capacity. The findings also demonstrate that variations in number of OST distractors can be used to alter sensitivity of the task.

Replay of Episodic Memories in the Rat.

Vivid episodic memories in people have been characterized as the replay of multiple unique events in sequential order [1-3]. The hippocampus plays a critical role in episodic memories in both people and rodents [2, 4-6]. Although rats remember multiple unique episodes [7, 8], it is currently unknown if animals "replay" episodic memories. Therefore, we developed an animal model of episodic memory replay. Here, we show that rats can remember a trial-unique stream of multiple episodes and the order in which these events occurred by engaging hippocampal-dependent episodic memory replay. We document that rats rely on episodic memory replay to remember the order of events rather than relying on non-episodic memories. Replay of episodic memories survives a long retention-interval challenge and interference from the memory of other events, which documents that replay is part of long-term episodic memory. The chemogenetic activating drug clozapine N-oxide (CNO), but not vehicle, reversibly impairs episodic memory replay in rats previously injected bilaterally in the hippocampus with a recombinant viral vector containing an inhibitory designer receptor exclusively activated by a designer drug (DREADD; AAV8-hSyn-hM4Di-mCherry). By contrast, two non-episodic memory assessments are unaffected by CNO, showing selectivity of this hippocampal-dependent impairment. Our approach provides an animal model of episodic memory replay, a process by which the rat searches its representations in episodic memory in sequential order to find information. Our findings using rats suggest that the ability to replay a stream of episodic memories is quite old in the evolutionary timescale.

Incidental spatial memory in the domestic dog (Canis familiaris).

We built upon previous work by Fujita et al. (2012, Animal Cognition, 15(6), 1055-1063) to create an experiment that investigated the presence of incidental memory for the spatial location of uneaten food in the domestic dog. Here, we dissociated potentially incidental spatial memory from the incidental memory for the characteristics of objects, in this case, food bowls. Eighteen household domestic dogs of various breeds and age were presented with four bowls. Each bowl contained either a novel object, treats the dog could consume, treats it could not consume, or it was left empty. Following a delay, the dogs returned to the laboratory and were presented with empty bowls in the same spatial orientation as the initial exposure and could move freely between bowls. This experiment required no previous training outside of basic obedience and so avoids the possibility that performance on the test was a conditioned response. We hypothesized that domestic dogs would be able to remember the location of uneaten food when presented with an unexpected memory test. We found that dogs in this study showed no evidence that they encoded spatial location in the absence of other cues that could be used to distinguish food bowls at specific locations. This suggests that dogs in previous experiments were more dependent on incidentally encoding the "what" and "in what" of this task than the "where," in the absence of features making each location distinct.

A test of the reward-contrast hypothesis.

Source memory, a facet of episodic memory, is the memory of the origin of information. Whereas source memory in rats is sustained for at least a week, spatial memory degraded after approximately a day. Different forgetting functions may suggest that two memory systems (source memory and spatial memory) are dissociated. However, in previous work, the two tasks used baiting conditions consisting of chocolate and chow flavors; notably, the source memory task used the relatively better flavor. Thus, according to the reward-contrast hypothesis, when chocolate and chow were presented within the same context (i.e., within a single radial maze trial), the chocolate location was more memorable than the chow location because of contrast. We tested the reward-contrast hypothesis using baiting configurations designed to produce reward-contrast. The reward-contrast hypothesis predicts that under these conditions, spatial memory will survive a 24-h retention interval. We documented elimination of spatial memory performance after a 24-h retention interval using a reward-contrast baiting pattern. These data suggest that reward contrast does not explain our earlier findings that source memory survives unusually long retention intervals.

Amnestic drugs in the odor span task: Effects of flunitrazepam, zolpidem and scopolamine.

The odor span task is an incrementing non-matching-to-sample procedure designed to provide an analysis of working memory capacity in rodents. The procedure takes place in an arena apparatus and rats are exposed to a series of odor stimuli in the form of scented lids with the selection of new stimuli reinforced. This procedure makes it possible to study drug effects as a function of the number of stimuli to remember. In the present study, the non-selective positive allosteric GABAA receptor modulator flunitrazepam impaired odor span performance at doses that did not affect a control odor discrimination. In contrast, the alpha-1 selective positive GABAA receptor modulator zolpidem and the cholinergic receptor antagonist scopolamine only impaired odor span at doses that produced more global impairment, including decreased accuracy in the control discrimination and increased response omissions in the both the odor span and control discrimination procedures. Even though the effects of flunitrazepam were selective to odor span performance, they did not depend on the number of stimuli to remember-the same degree of impairment occurred regardless of the memory load. These findings suggest that flunitrazepam interfered selectively with conditional discrimination performance rather than working memory and tentatively suggest that flunitrazepam's selective effects in the odor span task relative to the control odor discrimination are mediated by one or more non-alpha1 GABAA receptor subtypes.

The chemotherapeutic agent paclitaxel selectively impairs reversal learning while sparing prior learning, new learning and episodic memory.

Chemotherapy is widely used to treat patients with systemic cancer. The efficacy of cancer therapies is frequently undermined by adverse side effects that have a negative impact on the quality of life of cancer survivors. Cancer patients who receive chemotherapy often experience chemotherapy-induced cognitive impairment across a variety of domains including memory, learning, and attention. In the current study, the impact of paclitaxel, a taxane derived chemotherapeutic agent, on episodic memory, prior learning, new learning, and reversal learning were evaluated in rats. Neurogenesis was quantified post-treatment in the dentate gyrus of the same rats using immunostaining for 5-Bromo-2'-deoxyuridine (BrdU) and Ki67. Paclitaxel treatment selectively impaired reversal learning while sparing episodic memory, prior learning, and new learning. Furthermore, paclitaxel-treated rats showed decreases in markers of hippocampal cell proliferation, as measured by markers of cell proliferation assessed using immunostaining for Ki67 and BrdU. This work highlights the importance of using multiple measures of learning and memory to identify the pattern of impaired and spared aspects of chemotherapy-induced cognitive impairment.

Behavioral pharmacology of the odor span task: Effects of flunitrazepam, ketamine, methamphetamine and methylphenidate.

The Odor Span Task is an incrementing non-matching-to-sample procedure that permits the study of behavior under the control of multiple stimuli. Rats are exposed to a series of odor stimuli and selection of new stimuli is reinforced. Successful performance thus requires remembering which stimuli have previously been presented during a given session. This procedure has been frequently used in neurobiological studies as a rodent model of working memory; however, only a few studies have examined the effects of drugs on performance in this task. The present experiments explored the behavioral pharmacology of a modified version of the Odor Span Task by determining the effects of stimulant drugs methylphenidate and methamphetamine, NMDA antagonist ketamine, and positive GABAA modulator flunitrazepam. All four drugs produced dose-dependent impairment of performances on the Odor Span Task, but for methylphenidate and methamphetamine, these occurred only at doses that had similar effects on performance of a simple odor discrimination. Generally, these disruptions were based on omission of responding at the effective doses. The effects of ketamine and flunitrazepam were more selective in some rats. That is, some rats tested under flunitrazepam and ketamine showed decreases in accuracy on the Odor Span Task at doses that did not affect simple discrimination performance. These selective effects indicate disruption of within-session stimulus control. Overall, these findings support the potential of the Odor Span Task as a baseline for the behavioral pharmacological analysis of remembering.

Rats Remember Items in Context Using Episodic Memory.

Vivid episodic memories in people have been characterized as the replay of unique events in sequential order [1-3]. Animal models of episodic memory have successfully documented episodic memory of a single event (e.g., [4-8]). However, a fundamental feature of episodic memory in people is that it involves multiple events, and notably, episodic memory impairments in human diseases are not limited to a single event. Critically, it is not known whether animals remember many unique events using episodic memory. Here, we show that rats remember many unique events and the contexts in which the events occurred using episodic memory. We used an olfactory memory assessment in which new (but not old) odors were rewarded using 32 items. Rats were presented with 16 odors in one context and the same odors in a second context. To attain high accuracy, the rats needed to remember item in context because each odor was rewarded as a new item in each context. The demands on item-in-context memory were varied by assessing memory with 2, 3, 5, or 15 unpredictable transitions between contexts, and item-in-context memory survived a 45 min retention interval challenge. When the memory of item in context was put in conflict with non-episodic familiarity cues, rats relied on item in context using episodic memory. Our findings suggest that rats remember multiple unique events and the contexts in which these events occurred using episodic memory and support the view that rats may be used to model fundamental aspects of human cognition.

Emergent identity but not symmetry following successive olfactory discrimination training in rats.

The search for symmetry in nonhuman subjects has been successful in recent studies in pigeons (e.g., Urcuioli, 2008). The key to these successes has been the use of successive discrimination procedures and combined training on identity, as well as arbitrary, baseline relations. The present study was an effort to extend the findings and theoretical analysis developed by Urcuioli and his colleagues to rats using olfactory rather than visual stimuli. Experiment 1 was a systematic replication of Urcuioli's (2008) demonstration of symmetry in pigeons. Rats were exposed to unreinforced symmetry probes following training with two arbitrary and four identity conditional discriminations. Response rates on symmetry probe trials were low and provided little evidence for emergent symmetry in any of the seven rats tested. In Experiment 2, a separate group of six rats was trained on four identity relations and was then exposed to probe trials with four novel odor stimuli. Response rates were high on identity probe trials, and low on nonmatching probe trials. The similar patterns of responding on baseline and probe trials that were shown by most rats provided a demonstration of generalized identity matching. These findings suggest that the development of stimulus control topographies in rats with olfactory stimuli may differ from those that emerge in pigeons with visual stimuli. Urcuioli's (2008) theory has been highly successful in predicting conditions necessary for stimulus class formation in pigeons, but may not be sufficient to fully understand determinants of emergent behaviors in other nonhuman species.