Place cell firing cannot support navigation without intact septal circuits.

Though it has been known for over half a century that interference with the normal activity of septohippocampal neurons can abolish hippocampal theta rhythmicity, a definitive answer to the question of its function has remained elusive. To clarify the role of septal circuits and theta in location-specific activity of place cells and spatial behavior, three drugs were delivered to the medial septum of rats: Tetracaine, a local anesthetic; muscimol, a GABA-A agonist; and gabazine, a GABA-A antagonist. All three drugs disrupted normal oscillatory activity in the hippocampus. However, tetracaine and muscimol both reduced spatial firing and interfered with the rat's ability to navigate to a hidden goal. After gabazine, location-specific firing was preserved in the absence of theta, but rats were unable to accurately locate the hidden goal. These results indicate that theta is unnecessary for location-specific firing of hippocampal cells, and that place cell activity cannot support accurate navigation when septal circuits are disrupted.

Finding and Not Finding Rat Perirhinal Neuronal Responses to Novelty.

There is much evidence that the perirhinal cortex of both rats and monkeys is important for judging the relative familiarity of visual stimuli. In monkeys many studies have found that a proportion of perirhinal neurons respond more to novel than familiar stimuli. There are fewer studies of perirhinal neuronal responses in rats, and those studies based on exploration of objects, have raised into question the encoding of stimulus familiarity by rat perirhinal neurons. For this reason, recordings of single neuronal activity were made from the perirhinal cortex of rats so as to compare responsiveness to novel and familiar stimuli in two different behavioral situations. The first situation was based upon that used in "paired viewing" experiments that have established rat perirhinal differences in immediate early gene expression for novel and familiar visual stimuli displayed on computer monitors. The second situation was similar to that used in the spontaneous object recognition test that has been widely used to establish the involvement of rat perirhinal cortex in familiarity discrimination. In the first condition 30 (25%) of 120 perirhinal neurons were visually responsive; of these responsive neurons 19 (63%) responded significantly differently to novel and familiar stimuli. In the second condition eight (53%) of 15 perirhinal neurons changed activity significantly in the vicinity of objects (had "object fields"); however, for none (0%) of these was there a significant activity change related to the familiarity of an object, an incidence significantly lower than for the first condition. Possible reasons for the difference are discussed. It is argued that the failure to find recognition-related neuronal responses while exploring objects is related to its detectability by the measures used, rather than the absence of all such signals in perirhinal cortex. Indeed, as shown by the results, such signals are found when a different methodology is used. © 2016 The Authors Hippocampus Published by Wiley Periodicals, Inc.

Repetitive convulsant-induced seizures reduce the number but not precision of hippocampal place cells.

Repetitive one-per-day seizures induced in otherwise normal rats by the volatile convulsant flurothyl decrease the accuracy of locating a hidden goal without changing the mean location of goal selection. We now show that an 8-d series of such seizures degrades the spatial signal carried by the firing of hippocampal pyramidal cells and specifically reduces the information conveyed by the place cell subset of pyramidal cells. This degradation and a concomitant slowing of the hippocampal theta rhythm occur over time courses parallel to the development of the behavioral deficit and plausibly account for the impairment. The details of how pyramidal cell discharge weakens are, however, unexpected. Rather than a reduction in the precision of location-specific firing distributed evenly over all place cells, the number of place cells decreases with seizure number, although the remaining place cells remain quite intact. Thus, with serial seizures there is a cell-specific conversion of robust place cells to sporadically firing (<0.1 spike/s) "low-rate" cells as opposed to gradual loss of place cell resolution. This transformation occurs in the absence of significant changes in the discharge rate of hippocampal interneurons, suggesting that the decline in the number of place cells is not a simple matter of increased inhibitory tone. The cumulative transformation of place cells to low-rate cells by repetitive seizures may reflect a homeostatic, negative-feedback process.

Inhibition of protein kinase Mζ disrupts the stable spatial discharge of hippocampal place cells in a familiar environment.

It is widely held that spatial computations in the rodent hippocampus require the location-specific discharge of place cells that together form a stable cognitive map used to solve and perform spatial tasks. It is not known, however, if map stability requires persistent hippocampal synaptic strength changes that are vulnerable to blockade of protein kinase Mζ (PKMζ) phosphorylation activity, a manipulation that reverses hippocampal LTP and disrupts multiple forms of long-term memory. Here we report that acute intrahippocampal inhibition of PKMζ disrupts place cell activity in a familiar environment, where the map is expected to be stable. After this disruption, new, stable spatial firing patterns can later form, but the new and original maps are unrelated even though the rat is exposed to a constant environment. We therefore propose that the previously demonstrated erasure of stored spatial memory and the disruption of place cell firing are parallel effects of PKMζ blockade. We similarly propose that the known sparing of new spatial memory formation depends on the sparing of new map formation. On these bases, we argue that the loss of the map used to perform a practiced spatial task leads to behavioral performance deficits, and that synaptic plasticity maintained by PKMζ, which stabilizes the map, is essential for the proper expression of spatial memory.

Theta phase classification of interneurons in the hippocampal formation of freely moving rats.

Earlier work on freely moving rats classified neurons in Ammon's horn as pyramidal cells (including place cells) or interneurons (previously called "theta cells") based on temporal discharge correlates and waveform configurations, but the anatomical and biochemical diversity of interneurons suggests they may have other distinguishing characteristics. To explore this possibility, we made extracellular recordings as rats foraged for food in an open space, used accepted criteria to identify interneurons, and found two additional categorization methods. First, interneurons were separated into theta-modulated and theta-independent groups using spike autocorrelograms. Second, theta-modulated interneurons were further separated into four groups by the phase of the ∼8 Hz theta rhythm at which firing was most rapid. These phase groups resemble the four phase peak groups of five anatomically identified interneuron types (two with the same preferred phase) recorded during the slow (∼4 Hz) theta rhythm in urethane-anesthetized rats. We suggest that the similar number of peak phase groups in walking rats and urethane-anesthetized rats and the partial agreement between peak phase values reflect a similar organization of theta rhythm in both states, so that the discharge properties of anatomically identified interneurons can be described in freely moving rats. Interestingly, the average spatial firing precision of the interneuron classes does not differ significantly, suggesting that the strong location-specific firing of place cells may be due to segregated high- and low-precision interneuron ensembles rather than to one or more dedicated high-precision classes.

The presence of a second rat has only subtle effects on the location-specific firing of hippocampal place cells.

We compared the spatial firing properties of hippocampal place cells as a hungry rat foraged for randomly scattered food pellets in a familiar environment while it was by itself and while it shared the arena with a second rat that also was trained in the same task. Our goal was to determine if the hippocampal mapping system remained functional in the presence of the second rat, despite a strong initial tendency of the two animals to stay close together and despite the increased complexity of the sensory surroundings. We found that almost all place cell firing fields were only marginally changed by introducing the second rat. In particular, there was no evidence of the remapping characteristic of place cells in a sufficiently different novel environment. Instead, firing fields became somewhat less well organized and slightly weaker in the presence of the second rat. These second order changes were found to be distance dependent; the degradation of firing properties was maximal when the two rats were near each other. We conclude that signals in the hippocampal mapping system are affected to a small enough extent that accurate navigational is still possible when the environment is enriched in this realistic fashion.

Attention-like modulation of hippocampus place cell discharge.

Hippocampus place cell discharge is an important model system for understanding cognition, but evidence is missing that the place code is under the kind of dynamic attentional control characterized in primates as selective activation of one neural representation and suppression of another, competing representation. We investigated the apparent noise ("overdispersion") in the CA1 place code, hypothesizing that overdispersion results from discharge fluctuations as spatial attention alternates between distal cues and local/self-motion cues. The hypothesis predicts that: (1) preferential use of distal cues will decrease overdispersion; (2) global, attention-like states can be decoded from ensemble discharge such that both the discharge rates and the spatial firing patterns of individual cells will be distinct in the two states; (3) identifying attention-like states improves reconstructions of the rat's path from ensemble discharge. These predictions were confirmed, implying that a covert, dynamic attention-like process modulates discharge on a approximately 1 s time scale. We conclude the hippocampus place code is a dynamic representation of the spatial information in the immediate focus of attention.

Hippocampal place cell firing patterns can induce long-term synaptic plasticity in vitro.

In the hippocampus, synaptic strength between pyramidal cells is modifiable by NMDA receptor (NMDAR)-dependent long-term potentiation (LTP) and long-term depression (LTD), both of which require coincident presynaptic and postsynaptic activity. In vivo, many pyramidal cells exhibit location-specific activity patterns and are known as "place cells." The combination of these factors suggests that synaptic plasticity will be induced at synapses connecting place cells with overlapping firing fields, because such cells fire coincidentally when the rat is in a specific part of the environment. However, this prediction, which is important for models of how long-term synaptic plasticity can be used to encode space in the hippocampal network, has not been tested. To investigate this, action potential time series recorded simultaneously from place cells in freely moving rats were replayed concurrently into postsynaptic CA1 pyramidal cells and presynaptic inputs during perforated patch-clamp recordings from adult hippocampal slices. Place cell firing patterns induced large, pathway-specific, NMDAR-dependent LTP that was rapidly expressed within a few minutes. However, place-cell LTP was induced only if the two place cells had overlapping firing fields and if the cholinergic tone present in the hippocampus during exploration was restored by bath application of the cholinergic agonist carbachol. LTD was never observed in response to place cell firing patterns. Our findings demonstrate that spike patterns from hippocampal place cells can robustly induce NMDAR-dependent LTP, providing important evidence in support of a model in which spatial distance is encoded as the strength of synaptic connections between place cells.

Complementary spatial firing in place cell-interneuron pairs.

The hippocampal formation plays a pivotal role in representing the physical environment. While CA1 pyramidal cells display sharply tuned location-specific firing, the activity of many interneurons show weaker but significant spatial modulation. Although hippocampal interneurons were proposed to participate in the representation of space, their interplay with pyramidal cells in formation of spatial maps is not well understood. In this study, we investigated the spatial correlation between CA1 pyramidal cells and putative interneurons recorded concurrently in awake rats. Positively and negatively correlated pairs were found to be simultaneously present in the CA1 region. While pyramidal cell-interneuron pairs with positive spatial correlation showed similar firing maps, negative spatial correlation was often accompanied by complementary place maps, which could occur even in the presence of a monosynaptic excitation between the cells. Thus, location-specific firing increase of hippocampal interneurons is not necessarily a simple product of excitation by a pyramidal cell with a similarly positioned firing field. Based on our observation of pyramidal cells firing selectively in the low firing regions of interneurons, we speculate that the location-specific firing of place cells is partly determined by the location-specific decrease of interneuron activity that can release place cells from inhibition.

Discharge properties of hippocampal neurons during performance of a jump avoidance task.

We recorded single hippocampal cells while rats performed a jump avoidance task. In this task, a rat was dropped onto the metal floor of a 33 cm gray wooden cube and was given a mild electric shock if it did not jump up onto the box rim in <15 s. We found that many hippocampal pyramidal cells and most interneurons discharged preferentially at the drop, the jump, or on both events. By simultaneously recording the hippocampal EEG, we found that the discharge of most of the event-related pyramidal cells was modulated by the theta rhythm and moreover that discharge precessed with theta cycles in the same manner seen for pyramidal cells in their role as place cells. The elevations of firing rate at drop and jump were accompanied by increases in theta frequency. We conclude that many of the features of event-related discharge can be interpreted as being equivalent to the activity of place cells with firing fields above the box floor. Nevertheless, there are sufficient differences between expectations from place cells and observed activity to indicate that pyramidal cells may be able to signal events as well as location.

Recurrent seizures induce a reversible impairment in a spatial hidden goal task.

A major question concerning the learning and memory deficits characteristic of epilepsy is the relative importance of the initial insult that leads to recurrent, unprovoked seizures versus the seizures themselves. A related issue is whether seizure-induced cognitive decline is permanent or reversible when convulsions cease. To address these problems, adult rats were extensively trained in the "spatial accuracy task," a dry-land analog of the Morris water maze. This task allows the rat's estimate of the location of a hidden goal zone to be repeatedly measured within each behavioral session. One aim was to measure, in well-trained animals, the time course of any cognitive impairment caused by a daily flurothyl-induced generalized seizure over 11 days. A second aim was to look for possible recovery during 9 subsequent days with no seizures. We saw a cumulative degradation in spatial performance during the seizure days and reversal of the deficit after seizures were stopped such that performance returned to baseline. Interestingly, the rate of learning to an asymptote, the rate of performance decline during one-per-day seizures and the rate of relearning during the recovery period were all similar. Given that the hippocampus plays an important role in spatial memory and that it is the brain structure most vulnerable to abnormal excitation the implication is that the hippocampus remains essential for precise spatial navigation even after prolonged training in locating a fixed goal zone. Clinically, this finding questions the assumption that patients who experience seizures should return to a baseline cognitive level within hours.

Goal-related activity in hippocampal place cells.

Place cells are hippocampal neurons whose discharge is strongly related to a rat's location in its environment. The existence of place cells has led to the proposal that they are part of an integrated neural system dedicated to spatial navigation, an idea supported by the discovery of strong relationships between place cell activity and spatial problem solving. To further understand such relationships, we examined the discharge of place cells recorded while rats solved a place navigation task. We report that, in addition to having widely distributed firing fields, place cells also discharge selectively while the hungry rat waits in an unmarked goal location to release a food pellet. Such firing is not duplicated in other locations outside the main firing field even when the rat's behavior is constrained to be extremely similar to the behavior at the goal. We therefore propose that place cells provide both a geometric representation of the current environment and a reflection of the rat's expectancy that it is located correctly at the goal. This on-line feedback about a critical aspect of navigational performance is proposed to be signaled by the synchronous activity of the large fraction of place cells active at the goal. In combination with other (prefrontal) cells that provide coarse encoding of goal location, hippocampal place cells may therefore participate in a neural network allowing the rat to plan accurate trajectories in space.

Inhibition of kainate receptors reduces the frequency of hippocampal theta oscillations.

We investigated the role of kainate receptors in the generation of theta oscillations using (S)-1-(2-amino-2-carboxyethyl)-3-(2-carboxythiophene-3-yl-methyl)pyrimidine-2,4-dione (UBP304), a novel, potent and highly selective antagonist of GLU(K5)-containing kainate receptors. EEG and single-unit recordings were made from the dorsal hippocampus of awake, freely moving rats trained to forage for food. Bilateral intracerebroventricular injections of UBP304 (2.0 microl, two times; 2.08 mM) caused a clear (approximately 25%) reduction in theta frequency that was dissociable from behavioral effects of the drug. The locations of firing fields of principal cells in the hippocampal formation were generally preserved, but both field firing rates and the precision of field organization decreased. UBP304 lowered the frequency of the theta modulation of hippocampal interneuron discharge, accurately matching the reduced frequency of the theta field oscillation. UBP308 [(R)-1-(2-amino-2-carboxyethyl)-3-(2-carboxythiophene-3-yl-methyl)pyrimidine-2,4-dione], the inactive enantiomer of UBP304, caused none of these effects. Our results suggest that GLU(K5) receptors have an important role in modulating theta activity. In addition, the effects on cellular responses provide both insight into the mechanisms of theta pacing, and useful information for models of temporal coding.

Changes in goal selection induced by cue conflicts are in register with predictions from changes in place cell field locations.

In the cognitive mapping theory of hippocampal function, currently active place cells represent a rat's spatial location (J. O'Keefe & L. Nadel, 1978). A systematic shift of firing field locations should therefore produce a similar shift in a rat's judgment of its location. A. A. Fenton, G. Csizmadia, and R. U. Muller (2000a) recorded place cells in cylinders with 2 cue cards separated by 135 degrees . When the separation was changed, firing fields moved systematically, as described by a vector-field equation (A. A. Fenton, G. Csizmadia, & R. U. Muller, 2000b). Given this cohesive movement of firing fields, the mapping theory predicts that a rat's decisions about the location of an unmarked goal should move after card separation changes, as described by the vector-field equation. The authors tested this reasoning with a task in which the rat earned a food reward by pausing in a small, unmarked goal zone. When cues were shifted in the absence of reward, goal choice shifts were accurately predicted by the vector-field equation, providing strong support for the notion that a rat's judgment of its spatial location is intimately related to the across-cell discharge pattern of simultaneously active place cells.

Relationships between place cell firing fields and navigational decisions by rats.

This study examined the performance of spatial problems by rats when purely behavioral manipulations disturb the relationship between the place cell representation and the cues used to solve the problems. Place cells were recorded while rats performed a task in which they had to locate a goal in a gray cylinder. In the "far" task, the unmarked goal was displaced by a large fixed distance from a white card on the cylinder wall. In the "near" task, the unmarked goal was directly in front of the card. Finally, in the "cue" task the goal was marked by a black disk on the cylinder floor. Relationships between visible stimuli and place cell activity were manipulated by conducting either "hidden" (with the rat in its home cage) or "visible" (with the rat in the recording apparatus) rotations of the wall card and, when present, independent rotations of the black disk. Hidden card rotations generally caused equal firing field rotations, whereas visible card rotations often did not cause fields to move. In the far task, visible card rotations were associated with a strong decrease of correct responses in the card-referred goal area. Most rats tended to search the goal in the field-referred area. In the near task, visible card rotations were associated with a moderate decrease of performance, with rats searching the goal at the wall card. Finally, field placements had no effect on performance in the cue task. Thus, visible rotations tended to disrupt the relationship between firing fields and cues in all tasks but impaired performance only in the task that required map-based navigation. These results provide strong new evidence in favor of the spatial mapping theory of hippocampal function.

The effects on place cells of local scopolamine dialysis are mimicked by a mixture of two specific muscarinic antagonists.

Using a dialysis probe near CA1 hippocampal recording electrodes, we infused nonspecific (scopolamine) and specific (methoctramine, pirenzepine) antagonists of muscarinic cholinergic transmission to determine their effects on the positional firing properties of place cells. Both low (0.5 mM) and high (2.0 or 3.0 mM) scopolamine significantly decreased in-field firing rate, increased the ratio of out-of-field to in-field rate, and reduced the smoothness of rate maps, while tending to increase out-of-field rate. Thus, local nonspecific muscarinic blockade mimicked the effects seen with intracerebroventricular application, suggesting that blockade of receptors local to the recorded cells plays an essential role. Unexpectedly, dialysis of scopolamine reduced locomotor activity, again duplicating the effects of intracerebroventricular administration. Most effects of methoctramine (1.0 mM), which blocks presynaptic m2 and m4 receptors, were initially strong but then diminished over hours. Methoctramine produced a significant increase only in out/in ratio and out-of-field rate, whereas it tended to increase in-field rate and monotonically decrease smoothness. Pirenzepine (3.0 mM), which blocks postsynaptic m1 receptors, produced a significant increase only in out/in ratio, whereas it tended to increase out-of-field rate and decrease in-field rate; all these effects were monotonic with respect to time. A mixture of methoctramine plus pirenzepine recapitulated the place-cell effects of scopolamine, although neither the mixture nor its separate components affected behavior. We conclude that the effects of scopolamine on place cells likely result from a combination of blockade of postsynaptic m1 receptors, leading to reduced excitability, with blockade of presynaptic m2 and m4 receptors, leading to increased out-of-field firing.

Seizure-induced changes in place cell physiology: relationship to spatial memory.

Status epilepticus (SE) is a frequent neurological emergency associated with a significant risk of morbidity in survivors. Impairment of hippocampal-specific memory is a common and serious deficit occurring in many of the survivors. However, the pathophysiological basis of cognitive deficits after SE is not clear. To directly address the cellular concomitants of spatial memory impairment, we recorded the activity of place cells from CA1 in freely moving rats subjected to SE during early development and compared this activity to that in control rats. Place cells discharge rapidly only when the rat's head is in a cell-specific part of the environment called the "firing field." This firing field remains stable over time. Normal place cell function seems to be essential for stable spatial memory for the environment. We, therefore, compared place cell firing patterns with visual-spatial memory in the water maze in SE and control rats. Compared with controls, place cells from the SE rats were less precise and less stable. Concordantly, the water maze performance was also impaired. There was a close relationship between precision and stability of place cells and water maze performance. In contrast, a single, acute, chemically induced seizure produced cessation of place cell activity and spatial memory impairment in water maze performance that reversed within 24 hr. These results strongly bolster the idea that there is a relationship between abnormal place cells and spatial memory. Our findings also suggest that the defects in place cell and spatial memory after SE and acute chemically induced seizures result from different processes.

Place-cell impairment in glutamate receptor 2 mutant mice.

There is a strong correlation between Hebbian, NMDA receptor-dependent long-term potentiation (LTP), place-cell firing, and learning and memory. We made glutamate receptor 2 (GluR2) null mutant mice that show enhanced non-Hebbian LTP in hippocampal CA1 neurons and impaired performance in a spatial learning task. We concluded that in vivo hippocampal place cells of GluR2 mutant mice were functionally impaired because (1) only 22.6% of CA1 neurons showed place fields in GluR2 mutant mice, which was significantly lower than that (43.8%) in wild-type mice; (2) GluR2 mutant place fields were much less precise; and (3) GluR2 mutant place fields were extremely unstable. Our data suggest that place cells of GluR2 knock-out mice did not form robust place fields, and that enhanced non-Hebbian LTP might play a negative role in their formation.

Representation of objects in space by two classes of hippocampal pyramidal cells.

Humans can recognize and navigate in a room when its contents have been rearranged. Rats also adapt rapidly to movements of objects in a familiar environment. We therefore set out to investigate the neural machinery that underlies this capacity by further investigating the place cell-based map of the surroundings found in the rat hippocampus. We recorded from single CA1 pyramidal cells as rats foraged for food in a cylindrical arena (the room) containing a tall barrier (the furniture). Our main finding is a new class of cells that signal proximity to the barrier. If the barrier is fixed in position, these cells appear to be ordinary place cells. When, however, the barrier is moved, their activity moves equally and thereby conveys information about the barrier's position relative to the arena. When the barrier is removed, such cells stop firing, further suggesting they represent the barrier. Finally, if the barrier is put into a different arena where place cell activity is changed beyond recognition ("remapping"), these cells continue to discharge at the barrier. We also saw, in addition to barrier cells and place cells, a small number of cells whose activity seemed to require the barrier to be in a specific place in the environment. We conclude that barrier cells represent the location of the barrier in an environment-specific, place cell framework. The combined place + barrier cell activity thus mimics the current arrangement of the environment in an unexpectedly realistic fashion.

Impairment in long-term retention but not short-term performance on a water maze reversal task following hippocampal or mediodorsal striatal N-methyl-D-aspartate receptor blockade.

Male Long-Evans rats were injected with 32 ng/mul of the N-methyl-D-aspartate (NMDA) receptor antagonist 3-(2-carboxypiperazin-4-yl) propyl-1-phosphonic acid (CPP) or vehicle and trained to locate a hidden platform in a different location (reversal training) than used on the initial 4 days of training. Rats treated with vehicle or CPP into the dorsal hippocampus, basolateral amygdala, or mediodorsal striatum had similar latencies to locate the platform on the reversal day. Rats infused with CPP into the dorsal hippocampus or mediodorsal striatum failed to search preferentially in the novel location during a 24-hr, drug-free retention test, whereas all other groups searched preferentially in this location. Therefore, blocking dorsal hippocampal or mediodorsal striatal NMDA receptors selectively blocked long-term spatial retention without producing short-term performance deficits.