The effects of selective ibotenate lesions of the hippocampus on conditioned inhibition and extinction.

Previously, Solomon (1977) reported that aspiration lesions of the dorsal hippocampus in rabbits had no effect either on the acquisition of Pavlovian conditioned inhibition or on performance during a subsequent retardation test. The present experiment confirmed and extended these findings by showing that rats with ibotenate lesions of the complete hippocampus (the dorsal and ventral hippocampus and the dentate gyrus) were also unimpaired on the same types of tasks. Additional tests with the same rats showed that removing the hippocampus significantly impaired extinction of responding to a stimulus that had been previously trained with an appetitive unconditioned stimulus. The performance of the lesioned rats on a summation test was also marginally, but not significantly, different from that of controls. The data are discussed with reference to the idea that the hippocampus is involved with the formation of some, but not all, types of inhibitory associations.

The role of the entorhinal cortex in two forms of spatial learning and memory.

It is generally acknowledged that the rodent hippocampus plays an important role in spatial learning and memory. The importance of the entorhinal cortex (ERC), an area that is closely interconnected anatomically with the hippocampus, in these forms of learning is less clear cut. Recent studies using selective, fibre-sparing cytotoxic lesions have generated conflicting results, with some studies showing that spatial learning can proceed normally without the ERC, suggesting that this area is not required for normal hippocampal function. The present study compared cytotoxic and aspiration ERC lesions with both fimbria fornix (FFX) lesions and sham-operated controls on two spatial learning tasks which have repeatedly been shown to depend on the hippocampus. Both groups of ERC lesions were impaired during non-matching-to-place testing (rewarded alternation) on the elevated T-maze. However, neither of these lesions subsequently had any effect on the acquisition of a standard spatial reference memory task in the water maze. FFX lesions produced a robust and reliable impairment on both of these tasks. A second experiment confirmed that cytotoxic ERC lesions spared water maze learning but disrupted rewarded alternation on the T-maze, when the order of behavioural testing was reversed. These results confirm previous reports that ERC-lesioned animals are capable of spatial navigation in the water maze, suggesting that the ERC is not a prerequisite for normal hippocampal function in this task. The present demonstration that ERC lesions disrupt non-matching-to-place performance may, however, be consistent with the possibility that ERC lesions affect attentional mechanisms, for example, by increasing the sensitivity to recent reward history.

Contextual fear conditioning is disrupted by lesions of the subcortical, but not entorhinal, connections to the hippocampus.

Recent studies have questioned the importance of the entorhinal cortex (ERC) for normal hippocampal function. For example, fibre-sparing ERC lesions have been found to have no effect on spatial learning in the watermaze. There is also doubt as to the importance of the ERC for contextual fear conditioning, with previous studies having yielded conflicting results. In an attempt to resolve this issue, the present study compared aspiration and cytotoxic ERC lesioned rats, along with fimbria-fornix (FFX) lesioned animals and sham operated controls, on an unsignalled contextual fear conditioning paradigm. The results of the present study show that whereas lesions of the FFX disrupted contextual freezing, neither aspiration nor cytotoxic ERC lesions had any effect on this behaviour. Aspiration ERC lesioned rats, however, like FFX lesioned animals, did display hyperactivity prior to the delivery of footshock. These results suggest that whereas projections between the hippocampus and subcortical structures are important for normal levels of contextual freezing, projections from the entorhinal cortex are not essential.

The hippocampus and motivation revisited: appetite and activity.

After reviewing the available data regarding the various effects of manipulating (e.g. lesions, chemical or electrical stimulation) the hippocampal formation, Jarrard concluded that this structure likely played a role in motivated behaviors, specifically in general behavioral activation and incentive motivation. Since that time there have been technical advances in lesion techniques and conceptual advances in theories of motivation and learning. Here, we present more recent data that demonstrates the effects of hippocampal lesions on general activity, the utilization of interoceptive state cues, ingestive behaviors, and appetitive responding. We critically evaluate several theories of hippocampal function that have been proposed to explain these data, including the hippocampus as an inhibitor of general activation, as a processor of energy state signals and as a mediator of reward valuation. Finally, we propose that these findings may also be accounted for based on a role for the hippocampus in the learned inhibition of appetitive behaviors. We conclude that, while the specific mechanism of hippocampal involvement may not yet be determined, it is clear that this structure is involved in food-related behaviors and we caution researchers to consider this as a possible confound in studies of learning and memory processes.

Retrograde amnesia and consolidation: anatomical and lesion considerations.

The four papers in this issue of Hippocampus dealing with retrograde amnesia, together with relevant animal studies in the literature, are reviewed from the perspective of the anatomical location of the lesion and extent of damage to the brain. In order to evaluate the underlying damage in these and related prospective experimental studies, it is necessary to consider both the lesion techniques that were used as well as the care with which the resulting damage was determined. Both temporally graded and flat, ungraded retrograde amnesia have been reported, as well a lack of effects, following damage to structures in the medial temporal area. Most research has centered around damage to the hippocampus, but differences in selectivity of the lesions and behavioral testing procedures preclude any definite conclusions regarding the precise nature of the involvement of this structure. With a greater appreciation for the importance of the locus and extent of the damage, together with the kind of information being processed, it should be possible to obtain a better understanding of the neural substrates underlying retrograde amnesia.

Excitotoxic lesions of the pre- and parasubiculum disrupt object recognition and spatial memory processes.

Rats with bilateral ibotenic acid lesions centered on the pre- and parasubiculum and control rats were tested in a series of spatial memory and object recognition memory tasks. Lesioned rats were severely impaired relative to controls in both the reference and working memory versions of the water maze task and displayed a delay-dependent deficit in a delayed nonmatch to place procedure conducted in the T-maze. Lesioned rats also displayed reduced exploration in a novel environment, and performance was altered in an object recognition procedure as compared with the control group. These findings indicate that the pre- and parasubiculum plays an important role in the processing of both object recognition and spatial memory.

Reconsideration of the role of the hippocampus in learned inhibition.

The purpose of this article is to reconsider the role of the hippocampus in learning tasks that require suppression or prevention of memories or responses. This type of learning has generally been referred to as inhibitory learning. Although early theories proposed that the hippocampus was important for inhibitory learning, these ideas have generally fallen out of favor. However, new developments in the conceptual understanding of inhibition along with recent experimental evidence require that we review these conceptual changes with regard to hippocampal function. We review three general categories of 'inhibition' that have been used with reference to hippocampal function: neural inhibition, inhibition of attention and associative or learned inhibition. We then consider recent developments in the field of animal learning that call for changes in the early conceptualizations of learned inhibition. Specifically, current findings suggest that different types of learning paradigms can yield conditioned inhibitors that are embedded in different associative structures, at least some of which can co-exist with conditioned excitation. Next we proceed to review a number of recent experiments from our laboratory as well as others that encouraged renewed interest in the role for the hippocampus in inhibitory learning. We then conclude by considering some of the implications of the idea that the hippocampus is involved with performance of conditioned responses based on cues that are concurrently embedded in inhibitory and excitatory associations.

Selective lesions of the entorhinal cortex, the hippocampus, or the fimbria-fornix in rats: a comparison of effects on spontaneous and amphetamine-induced locomotion.

Using adult Long-Evans male rats, this experiment compared spontaneous (assessed 15 days and 4.5 months after surgery) and amphetamine-induced (assessed from 4.5 months after surgery onwards; 1 mg/kg, i.p., ten injections, 48 h apart) locomotor activity following N-methyl-D-aspartate lesions of the entorhinal cortex, electrolytic lesions of the fimbria-fornix, or ibotenate lesions of the hippocampus. Sham-operated rats were used as controls. Hippocampal and fimbria-fornix lesions, but not entorhinal-cortex lesions induced diurnal and nocturnal hyperactivity, which was attenuated over time, but only in rats with fimbria-fornix lesions. Amphetamine-induced hyperlocomotion was assessed in a familiar environment. Lesions of the entorhinal cortex potentiated the locomotor effects of amphetamine, but not lesions of the hippocampus or interruption of the axons in the fimbria-fornix pathway. Sensitization appeared to be decreased by fimbria-fornix lesions and to be prevented by hippocampal lesions. Rats with entorhinal-cortex lesions behaved as if they had already been sensitized by the lesion. These results clearly show that lesions of the fimbria-fornix, the hippocampus, and of the entorhinal cortex have different effects on spontaneous and amphetamine-induced hyperactivity, as they also have on learning and memory tasks.

Hippocampectomized rats are impaired in homing by path integration.

Theoretical, behavioral, and electrophysiologic evidence suggests that the hippocampal formation may play a role in path integration, a form of spatial navigation in which an animal can return to a starting point by integrating self-movement cues generated on its outward journey. The present study examined whether the hippocampus (Ammon's horn and the dentate gyrus) is involved in this form of spatial behavior. Control rats and rats with selective ibotenic acid lesions of the hippocampus were tested in a foraging task in which they retrieved large food pellets from an open field, which when found, they carried to a refuge for consumption. The experiments measured the rats' homing accuracy, returning to the starting location, under conditions in which visual, surface, and self-movement cues; surface and self-movement cues; or only self-movement cues were available. Although both control rats and rats without a hippocampus could use visual and surface cues, only control rats appeared to be able to use self-movement cues. The finding that hippocampal rats are impaired under conditions requiring the use of self-movement cues suggests that the hippocampus plays an essential role in path integration.

Effects of ibotenate hippocampal and extrahippocampal destruction on delayed-match and -nonmatch-to-sample behavior in rats.

The effects of ibotenate lesions of the hippocampus (HIPP) or hippocampus plus collateral damage to extrahippocampal structures (HCX) were investigated in rats trained to criterion on spatial versions of either a delayed-match (DMS) or delayed-nonmatch-to-sample (DNMS) task. After recovery from surgery, animals were retrained at "0" sec delays, then assessed at 0-30 sec delays for 15 d, retrained again at 0 sec delays, and retested for another 25 d on 0-30 sec delays. Pretrained HIPP-lesioned animals showed marked delay-dependent deficits in both tasks that never recovered. Detailed examination of within- and between-trial performance factors, including changes in response preferences, length of previous trial delay, and sequential dependencies, revealed important factors operating in lesioned animals that were either absent or insignificant before the lesion. Pretrained HCX-lesioned animals showed deficits similar to those of HIPP animals, with the noticeable exception of a strong "recency" influence of the previous trial. Another group of HIPP- and HCX-lesioned animals trained on the tasks after the lesion showed reduced impairments of the type described above, suggesting that extrahippocampal structures trained after the lesion can assume the role of the hippocampus to some degree. The findings indicate that both the type of lesion and the previous history of the animal determine the postlesion DMS and DNMS performance of animals suffering damage to the hippocampus and/or related structures.

Fimbria-fornix vs selective hippocampal lesions in rats: effects on locomotor activity and spatial learning and memory.

The behavioral effects of interrupting the axons that pass in the fimbria and dorsal fornix were compared with the effects of selective removal of the cells that comprise the hippocampus with ibotenic acid. Starting 4.5 months after surgery, lesioned and control rats were (i) trained in both the Morris water maze and the eight-arm radial maze using protocols that placed an emphasis on either working memory (WM) or reference memory (RM) and (ii) tested for locomotor activity in the home cage. In comparison to sham-operated rats, the rats from both lesion groups were impaired in most learning/memory tasks, but there were some interesting differences between the two lesioned groups. When compared to rats with fimbria-fornix lesions (FIFX rats), hippocampal rats (HIPP rats) were slower in learning to swim to a visible platform and showed a greater impairment than FIFX rats in the radial-maze task when the testing procedure required the utilization of RM and WM in a more demanding WM task. In the test of locomotor activity, FIFX and control rats did not differ, but HIPP rats were more active than the rats in both other groups. The pattern of results obtained after a 4.5-month recovery period support the following general conclusions. (1) While there are some similarities in the effects on behavior of interrupting the axons in the fimbria-fornix compared to removing the hippocampus, there are some important differences. (2) From the findings that are available, a possible explanation to account for the difference between FIFX and HIPP rats is that the 4.5-month survival time permitted some recovery in the group of rats with FIFX lesions. (3) While it is well known that the Morris water maze and the radial-arm maze tasks provide useful measures of spatial learning and memory processes, our results suggest that the information provided by the two spatial learning tasks may differ in important respects.

Behaviors and neurodegeneration induced by two blockers of K+ channels, the mast cell degranulating peptide and Dendrotoxin I.

Both the Mast Cell Degranulating (MCD) peptide and Dendrotoxin I (DTX(I)), two blockers of fast activation and slowly inactivating K+ channels, induced epileptiform seizures and brain damage after intracerebroventricular injection (200 pmol) in Sprague-Dawley rats. A considerable variation in the response of the rats was observed for each toxin. The neurodegeneration included the hippocampal formation, subiculum, septum, amygdala, and the cerebellum for both toxins, and the neocortex and anterior thalamic nuclei exclusively for DTX(I) treatment.

Effects of postoperative housing conditions on functional recovery in rats with lesions of the hippocampus, subiculum, or entorhinal cortex.

In order to study the effects of differential housing conditions on recovery from damage to different components of the hippocampal formation, 85 rats received bilateral lesions of the hippocampus, entorhinal cortex, or subiculum or sham surgery and then were housed for 30 days in either an enriched environment or an impoverished environment. Rats were subsequently tested on a battery of tasks for assessing locomotor activity in their home cage, reactivity to novelty, spatial working and reference memory in the Morris water maze, and learning in the Hebb-Williams maze. Rats with the hippocampus removed showed impairments in most of the tasks we used (home-cage and novelty-induced locomotor activity, water maze, and Hebb-Williams maze). Most of the deficits induced by lesions to the entorhinal cortex were similar to those induced by the removal of the hippocampus. Some differences appear to be among the deficits induced by the lesions of these structures when assessing the home-cage locomotor activity, the reactions to novelty, and one aspect of the Hebb-Williams maze learning. Lesions to the subiculum induced only an impairment in the probe trial of the water-maze task. Confirming and extending previous findings in rats with various (but nonexcitotoxic) lesions of the hippocampus, an enriched environment had a beneficial effect on several of the deficits observed in the tasks we used. Further, only the rats with hippocampal lesions benefitted from having been housed in the enriched environment. However, their facilitated recovery was not observed in all tasks. After damage to different components of the hippocampal formation, the beneficial effects induced by the enriched housing conditions were shown to be both lesion-locus- and task-dependent.

Facilitation of conditioned odor aversion by entorhinal cortex lesions in the rat.

This study examined the role of the entorhinal cortex (EC) in conditioned odor aversion learning (COA). Lateral EC lesions did not impair but rather facilitated COA. In the experiments the delay separating the odor cue presentation from the subsequent toxicosis was varied during acquisition. EC-lesioned rats demonstrated COA for delays up to 2 hr, whereas sham-operated rats displayed COA only if toxicosis immediately followed the odor cue. This facilitation was not dependent on the intensity of the odor and corresponded to a facilitated long-delay learning. EC lesion did not affect conditioned taste aversion, confirming that the facilitation effect does not correspond to a general facilitation of conditioned aversion learning. Taken together, these results indicate that the removal of the EC may allow odor-toxicosis associations across longer delays by extending the duration of the olfactory trace.

Is the lack of protein F1/GAP-43 mRNA in granule cells target-dependent?

Protein F1/GAP-43 is differentially expressed in brain with high levels present in regions associated with memory functions. However, in hippocampus the granule cells lack F1/GAP-43 expression. To determine if this lack of expression is due to inhibitory signals from the target cells, we selectively destroyed CA3 pyramidal cells unilaterally using microinjections of excitotoxins. Kainate lesions induced F1/GAP-43 mRNA expression bilaterally in granule cells at 24 h post-injection. Since the induction contralateral to the lesion was not due to loss of target cells, that induction may be ascribed to consequences of seizure activity. However, F1/GAP-43 mRNA hybridization decreased by 3 d post-lesion and was at background levels by 6 d, indicating that the lack of F1/GAP-43 expression in granule cells is restored despite a lack of target neurons. Unilateral lesions of CA3 cells using ibotenate, which are not as complete as kainate but do not cause seizures, did not induce F1/GAP-43 mRNA in granule cells on either the contralateral or, in 4 of 5 cases, the ipsilateral side. Taken together, these data suggest that the CA3 target is not essential for the absence of F1/GAP-43 expression in granule cells. To compare the extent of damage caused by the lesions, we investigated the location of astrocytes undergoing reactive gliosis, employing as a reporter glial fibrillary acidic protein (GFAP) gene expression. After both kainate and ibotenate injections GFAP hybridization increased in the lesioned area as well as in the contralateral hippocampus. These results indicate that injections of kainate, and possibly ibotenate to a lesser extent, may affect behavior not only by damaging cells at the injection site, but also by altering gene expression in cells at distant sites.

Evidence for extrahippocampal involvement in place learning and hippocampal involvement in path integration.

Although there is a good deal of evidence that animals require the hippocampus for learning place responses, animals with damage to the afferent and efferent fibers coursing through the fimbria-fornix have been shown to acquire a place response. This finding suggests either that the cells of the hippocampus proper (CA1-4 and dentate gyrus), via their connections to the temporal lobe, can mediate place learning or that some extrahippocampal structure is sufficient. We examined this question using rats with ibotenic acid lesions of the cells of the hippocampus. Rats were pretrained to swim to a visible platform and then given probe trials on which the visible platform was removed. Video and kinematic analyses showed that the hippocampal rats expected to find the platform at its previous location because they swam directly to that location and paused and turned at that location after the platform was removed. Additional tests confirmed that they had learned a place response. There were, however, abnormalities in their swimming patterns, and despite having acquired one place response, they did not then acquire new place responses when only the hidden platform training procedure was used. These results demonstrate that place learning can be acquired by rats in which the hippocampus proper is removed. Contrasts between conditions in which hippocampal rats acquire a place response and conditions in which they fail suggests that the hippocampus may serve as an on line system for monitoring movement and integrating movement paths.

What does the hippocampus really do?

Much of the evidence used to implicate the hippocampus in learning and memory has been obtained from clinical cases and/or experimental studies with animals where the damage is extensive and includes more than just the hippocampus. When the damage is limited to the cells that comprise the hippocampus (CA1-CA3 pyramidal cells, hilar and granule cells in the dentate gyrus) the effect on behavior in the rat is more limited than what is usually reported. Selective, axon-sparing ibotenic acid lesions of the hippocampus were used in the experiments that are reviewed to study the effects of removing the hippocampus on: (1) the acquisition of spatial and non-spatial information; (2) complex, non-spatial representational learning; and (3) acquisition and utilization of contextual information. The results indicated that rats with the hippocampus removed were impaired on those tasks that require the utilization of spatial and contextual information but performed like controls in learning about and handling (even complex) non-spatial information. Future research utilizing selective lesions of the hippocampus and sensitive behavioral testing techniques should help clarify the extent to which the impairments in the acquisition of spatial information and the ability to utilize contextual, background cues can be reduced to a single, underlying learning process.

Similarities vs. differences in place learning and circadian activity in rats after fimbria-fornix section or ibotenate removal of hippocampal cells.

Damage to either the fimbria-fornix or to the hippocampus can produce a deficit in spatial behavior and change in locomotor activity but the extent to which the two kinds of damage are comparable is not known. Here we contrasted the effects of cathodal sections of the fimbria-fornix with ibotenic acid lesions of the cells of the hippocampus (Ammon's horn and the dentate gyrus) on place learning in a swimming pool and on circadian activity. Rats in both ablation groups were impaired relative to control rats in learning a single place response but they did acquire the response as measured by swim latencies, errors, and by enhanced searching on probe trials. They were also more active than the control group on the test of activity. Nevertheless, the fimbria-fornix group was initially more impaired on learning and was more active than the hippocampal group. Analysis of the strategies used in learning indicated that the lesion groups were very similar to each other but different from the control group especially in that at asymptotic performance, rats in both lesion groups made rather tight loops as they swam toward the platform. This strategy likely contributed to the greater proportion of time they spent swimming in the correct quadrant on the subsequent probe trial. These findings confirm that rats with fimbria-fornix or hippocampal damage display impairments in place learning and are hyperactive but also show that there are lesion differences. The results are discussed with respect to the relative effectiveness of the lesions and the possibility that fibers in the fimbria-fornix may mediate some functions that are not attributable to the hippocampus.

Silver impregnation reveals neuronal damage in cingulate cortex following 4 VO ischaemia in the rat.

An intriguing feature of global ischaemic cell loss is the sensitivity of certain neuronal populations and the relative resistance of others. Silver impregnation was used to ascertain the pattern and extent of cell loss following 15 min 4 VO ischaemia in the rat. Cell loss was observed primarily in the CA1 region of the hippocampus, as assessed by both cresyl violet and silver stains. However, degenerating neurones were most readily identifiable when impregnated with silver, and additional regions of neuronal loss were selectively revealed by silver staining in the hippocampal hilar region, dorsolateral striatum, neocortex and cingulate cortex. Damage to cingulate is a hitherto unreported consequence of 4 VO global ischaemia. This novel finding may have implications for ischaemic brain-behaviour relationships.

Selective excitotoxic pathology in the rat hippocampus.

The pattern of cell loss and neuronal degeneration resulting from multiple microinjections of N-methyl-D-aspartate (NMDA), ibotenate (IBO), quisqualate (QUIS), and kainate (KA) into hippocampus was studied, together with the protection provided by the NMDA antagonist 3-(+/-)-2-carboxypiperazin-4-yl-propyl-1-phosphonate (CPP). Histological evaluation was carried out after 7 days of survival. NMDA and IBO resulted in an extensive loss of all cells in the hippocampus including dentate gyrus, hilar cells, and CA3-CA1 pyramidal cells, but there was an absence of damage to areas and structures outside hippocampus. After QUIS and KA injections the hippocampal damage was limited to hilar cells in the dentate gyrus, CA3 pyramidal cells, and partial loss of CA1 cells; there was extensive extrahippocampal damage including entorhinal cortex, amygdala, layers III, V, and VI of ventral neocortex, olfactory areas, and various thalamic nuclei. CPP provided almost complete protection from the effects of intrahippocampal injections of NMDA and IBO, but did not affect the hippocampal cell loss found after QUIS and KA (with the exception of minor protection of some CA1 cells). CPP protected most extrahippocampal sites from the damage resulting from QUIS and KA, indicating that such excitotoxic cell death is indirect and involves NMDA receptor activation by an endogenous agent. The use of multiple microinjections as opposed to single injections allows a clearer interpretation of selective excitotoxic vulnerability.