Showing papers in "Hippocampus in 1995"

Perirhinal and postrhinal cortices of the rat: A review of the neuroanatomical literature and comparison with findings from the monkey brain

Abstract: This review is prompted by recent findings that the perirhinal and parahippocampal cortices in the monkey brain are important components of the medial temporal lobe memory system. Given the potential impor- tance of the comparable regions to memory function in the rat brain, it is surprising that so little is known about their neuroanatomy. In fact, there are no comprehensive studies of the borders, cytoarchitecture, or connec- tions of the cortical regions surrounding the posterior portion of the rhi- nal sulcus in the rat. This review is meant to summarize the current state of our knowledge regarding these regions in the rat brain. Based on exist- ing data and our own observations, a new terminology is introduced that retains the term perirhinal cortex for the rostral portion of the region and renames the caudal portion the postrhinal cortex. Issues of continuing un- certainty are highlighted, and information gleaned from the monkey liter- ature is used to predict what anatomical traits the rat perirhinal region might demonstrate upon further examination. To the extent possible with available data, the similarities and differences of the rat and monkey perirhi- nal, postrhinal, and parahippocampal regions are evaluated.

Topographically specific hippocampal projections target functionally distinct prefrontal areas in the rhesus monkey

Abstract: The sources of ipsilateral projections from the hippocampal formation, the presubiculum, area 29a-c, and parasubiculum to medial, orbital, and lateral prefrontal cortices were studied with retrograde tracers in 27 rhesus monkeys. Labeled neurons within the hippocampal formation (CA1, CA1', prosubiculum, and subiculum) were found rostrally, although some were noted throughout the entire rostrocaudal extent of the hippocampal formation. Most labeled neurons in the hippocampal formation projected to medial prefrontal cortices, followed by orbital areas. In addition, there were differences in the topography of afferent neurons projecting to medial when compared with orbital cortices. Labeled neurons innervating medial cortices were found mainly in the CA1' and CA1 fields rostrally, but originated in the subicular fields caudally. In contrast, labeled neurons which innervated orbital cortices were considerably more focal, emanating from the same relative position within a field throughout the rostrocaudal extent of the hippocampal formation. In marked contrast to the pattern of projection to medial and orbital prefrontal cortices, lateral prefrontal areas received projections from only a few labeled neurons found mostly in the subicular fields. Lateral prefrontal cortices received the most robust projections from the presubiculum and the supracallosal area 29a-c. Orbital, and to a lesser extent medial, prefrontal areas received projections from a smaller but significant number of neurons from the presubiculum and area 29a-c. Only a few labeled neurons were found in the parasubiculum, and most projected to medial prefrontal areas. The results suggest that functionally distinct prefrontal cortices receive projections from different components of the hippocampal region. Medial and orbital prefrontal cortices may have a role in long-term mnemonic processes similar to those associated with the hippocampal formation with which they are linked. Moreover, the preponderance of projection neurons from the hippocampal formation innervating medial when compared with orbital prefrontal areas followed the opposite trend from what we had observed previously for the amygdala (Barbas and De Olmos [1990] (J Comp Neurol 301:1-23). Thus, the hippocampal formation, associated with mnemonic processes, targets predominantly medial prefrontal cortices, whereas the amygdala, associated with emotional aspects of memory, issues robust projections to orbital limbic cortices. Lateral prefrontal cortices receive robust projections from the presubiculum and area 29a-c and sparse projections from the hippocampal formation. These findings are consistent with the idea that the role of lateral prefrontal cortices in memory is distinct from that of either medial or orbital cortices. The results suggest that signals from functionally distinct limbic structures to some extent follow parallel pathways to functionally distinct prefrontal cortices.

Configural association theory and the hippocampal formation: an appraisal and reconfiguration.

Abstract: Sutherland and Rudy ([1989] Psychobiology 17:129–144) proposed that the hippocampal system is critical to normal learning and memory because of its function as the central part of a configural association system. This system constructs a unique representation of the joint occurrence of the independent elements of a compund. There is evidence consistent with the theory's predictions, however, there also are data that unambiguously demonstrate that, under some conditions, animals lacking an intact hippocampal system acquire configural associations. Thus, Sutherland and Rudy's fundamental assumption cannot be correct. To integrate the supporting and contradictory data, we propose two simple modifications of our position: (1) The critical neural system for configural associations is in cortical circuitry outside the hippocampus, and (2) the output from the hippocampal formation contributes to configural processing by selectively enhancing, thereby making more salient, cortical units representing stimulus conjunctions. This enhancement has two important effects: (1) It decreases the similarity between the configural units representing the co-occurrence of cues and the units representing the cues, and (2) it increases the rate at which the configural units can acquire associative strength. The modified theory explains why damage to the hippocampal formation only impairs learning on a subset of nonlinear discrimination problems. It also integrates recent data on the effects of hippocampal formation damage on conditioning involving context cues and makes novel predictions about performance on nonlinear discrimination problems and place learning. © 1995 Wiley-Liss, Inc.

Intracellular correlates of hippocampal theta rhythm in identified pyramidal cells, granule cells, and basket cells

Abstract: The cellular-synaptic generation of rhythmic slow activity (RSA or theta) in the hippocampus has been investigated by intracellular recording from principal cells and basket cells in anesthetized rats. In addition, the voltage-, coherence-, and phase versus depth profiles were examined by simultaneously recording field activity at 16 sites in the intact rat, during urethane anesthesia, and after bilateral entorhinal cortex lesion. In the extracellular experiments the large peak of theta at the hippocampal fissure was attenuated by urethane anesthesia and abolished by entorhinal cortex lesion. The phase versus depth profiles were similar during urethane anesthesia and following entorhinal cortex lesion but distinctly different in the intact, awake rat. These observations suggest that dendritic currents underlying theta in the awake rat may not be revealed under urethane anesthesia. The frequency of theta-related membrane potential oscillation was voltage-independent in pyramidal neurons, granule cells, and basket cells. On the other hand, the phase and amplitude of intracellular theta were voltage-dependent in all three cell types with an almost complete phase reversal at chloride equilibrium potential in pyramidal cells and basket cells. At strong depolarization levels (less than 30 mV) pyramidal cells emitted calcium spike oscillations, phase-locked to theta. Basket cells possessed the most regular membrane oscillations of the three cell types. All neurons of this study were verified by intracellular injection of biocytin. The observations provide direct evidence that theta-related rhythmic hyper-polarization of principal cells is brought about by the rhythmically discharging basket neurons.(ABSTRACT TRUNCATED AT 250 WORDS)

Selective damage to the hippocampal region blocks long‐term retention of a natural and nonspatial stimulus‐stimulus association

Abstract: Normal rats rapidly acquire and remember associations between nonspatial stimuli as expressed in the social transmission of food preferences. In the present study, rats with selective neurotoxic lesions including all subdivisions of the hippocampal region (hippocampus proper, dentate gyrus, and subiculum) normally acquired and briefly retained the food odor association as demonstrated by intact memory immediately after social training. However, long-term memory in these animals was severly impaired in contrast to strong 24-h retention by intact rats. More selective lesions to the hippocampus proper plus dentate gyrus alone, or the subiculum alone had no effect on memory at either test interval. These finding indicate that the hippocampal region is required for long-term retention of a nonspatial form of natural memory. © 1995 Wiley-Liss, Inc.

Relational features of acetylcholine, noradrenaline, serotonin and GABA axon terminals in the stratum radiatum of adult rat hippocampus (CA1).

Abstract: In a well-defined sector of adult rat hippocampus (CA1, stratum radiatum), the ultrastructural features of acetylcholine (ACh), noradrenaline (NA), serotonin (5-HT) and GABA axon terminals (varicosities) were compared by electron microscopy after immunostaining with antibodies against choline acetyltransferase, NA, 5-HT and glutamic acid decarboxylase. Approximately 100 sectional profiles of each type were analyzed for size, presence of a synaptic membrane specialization (synaptic incidence) and composition of the microenvironment. An equivalent number of immunonegative varicosity profiles selected at random from the same micrographs were similarly examined. ACh, NA and 5-HT varicosity profiles were of comparable size, and significantly smaller than GABA profiles. They exhibited a low frequency of junctional specialization, amounting to 7%, 15% and 21%, respectively, when extrapolated to the whole volume of these terminals. In contrast, GABA varicosities appeared entirely synaptic. The ACh, NA and 5-HT varicosities also differed from their GABA counterparts in being juxtaposed to a greater number of unlabeled axonal varicosities and a lower number of dendritic branches. In addition, the microenvironment of immunostained terminals showed a much lower number of dendritic spines than that of immunonegative varicosities. This latter finding was viewed as another indication that predominantly asynaptic varicosities do not maintain particular relationships with their immediate surround. It was also concluded that volume transmission represents a major mode of transmission for ACh, NA and 5-HT in adult rat hippocampus, thus contributing to the properties and functions assigned to these transmitters in this part of brain. © 1995 Wiley-Liss, Inc.

View-responsive neurons in the primate hippocampal complex.

Abstract: Recordings were made from single neurons in the hippocampus and parahippocampal gyrus while macaques were moved on a platform mounted on a free-moving robot or on wheels in a cue-controlled 2 m x 2 m x 2 m environment, in order to investigate the representation of space and of spatial memory in the primate hippocampus. The test conditions allowed factors that might account for spatial firing of the cells, including the spatial location where the monkey looked, the place were the monkey was, and the head direction of the monkey, to be identified. The responses of some ("view") neurons depended on where the monkey was looking in the environment, but not on the place of the monkey in the environment. The responses of one other neuron depended on a combination of where the monkey was facing and his place in the test chamber. The response of view-dependent neurons was affected by occlusion of the visual field. It was possible to show for one neuron that its "view" response rotated with rotation of the test chamber. Some neurons responded to a combination of whole-body motion and view or place, and one neuron responded in relation to whole-body movement to a particular place. One neuron responded depending on the place where the monkey was in the environment and relatively independently of view. The representations of space provided by hippocampal view-responsive neurons may be useful in forming memories of spatial environments (for example, of where an object has been seen and of where the monkey is as defined by seen views) and, together with whole-body motion cells, in remembering trajectories through environments, which is of use, for example, in short range spatial navigation.

Brain temperature and hippocampal function

Abstract: Even though homeothermic animals regulate the body temperature, fluctuations up to 2-3 degrees C may occur during physiological conditions. In many species, including the rat, a similar variation can be measured in the brain temperature. Such changes are expressed throughout the brain with a preserved gradient between the warmer basal and cooler dorsal parts. In spite of these recordable physiological changes, spatial learning is quite robust, in that it occurs at brain temperatures between 30 and 39 degrees C. Even drastic cooling (to below 15 degrees C) fails to affect consolidation or storage of information when the animal is tested after rewarming. The physiological temperature fluctuations have significant consequences for electrophysiological responses in the brain. Various bioelectrical signals are more sensitive during warming, axonal conduction is speeded up, and stimulus-elicited transmitter release becomes faster and more synchronized. Action potentials have shorter rise and decay times in warm conditions, and the amplitude becomes slightly smaller. Population responses are differently affected by these changes. Dentate field potentials in response to stimulation of perforant-path fibers appear with shorter latency in warm conditions, and the rate of rise in the field EPSP is increased. Paradoxically, the amplitude of the population spike is reduced. This is due to a combination of reduced amplitude of individual action potentials and reduced efficiency of the summation of groups of action potentials. Due to the large effects of temperature on hippocampal field potentials, it is mandatory that brain temperature changes are monitored and/or controlled whenever such responses are recorded in freely moving and anesthetized animals.

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

Abstract: 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.

Connection matrix of the hippocampal formation: I. The dentate gyrus

Abstract: The hippocampal formation presents a special opportunity for realistic neural modeling since its structure, connectivity, and physiology are better understood than that of other cortical components. A review of the quantitative neuroanatomy of the rodent dentate gyrus (DG) is presented in the context of the development of a computational model of its connectivity. The DG is a three-layered folded sheet of neural tissue. This sheet is represented as a rectangle, having a surface area of 37 mm2 and a septotemporal length of 12 mm. Points, representing cell somata, are distributed in the model rectangle in a roughly uniform fashion. Synaptic connectivity is generated by assigning each presynaptic cell a spatial zone representing its axonal arbor. For each postsynaptic cell, a list of potential presynaptic cells is compiled, based on which arbor zones the given postsynaptic cell falls within. An appropriate number of presynaptic inputs are then selected at random. The principal cells of the DG, the granule cells, are represented in the model, as are non-principal cells, including basket cells, chandelier cells, mossy cells, and GABAergic peptidergic polymorphic (GPP) cells. The neurons of layer II of the entorhinal cortex are included also. The DG receives its main extrinsic input from these cells via the perforant path. The basket cells, chandelier cells, and GPP cells receive perforant path and granule cell input and exert both feedforward and feedback inhibition onto the granule cells. Mossy cells receive converging input from granule cells and send their output back primarily to distant septotemporal levels, where they contact both granule cells and non-principal cells. To permit numerical simulations, the model must be scaled down while preserving its anatomical structure. A variety of methods for doing this exist. Hippocampal allometry provides valuable clues in this regard.

Place cells recorded in the parasubiculum of freely moving rats.

Abstract: Previous studies have identified neurons in the hippocampus, subiculum, and entorhinal cortex which discharge as a function of the animal's location in the environment In contrast, neurons in the postsubiculum and anterior thalamic nucleus discharge as a function of the animal's head direction in the horizontal plane, independent of its behavior and location in the environment Because the parasubiculum (PaS) has extensive connections, either directly or indirectly, with these structures, it is centrally located to influence the neuronal activity in these areas This study was therefore designed to determine the types of behavioral and spatial correlates in neurons from the PaS Single unit recordings were conducted in the PaS of freely moving rats trained to retrieve food pellets thrown randomly into a cylindrical apparatus A total of 103% of the cells were classified as place cells because they discharged in relation to the animal's location in the cylinder A large percentage of cells (414%) were classified as theta cells The remaining cells had nondiscernable behavioral correlates Quantitative analysis of the firing rate maps for the place cells showed they had higher levels of background activity and contained larger firing fields than values reported previously for hippocampal place cells Directional analysis showed that only three out of 16 cells contained a secondary directional correlate; the firing rate for the remaining cells was not affected by the animal's directional heading within the firing field A time shift analysis, which shifted the spike time series relative to the animal location series, was conducted to determine whether the quality of the location-specific firing could be improved The time shifts for three different spatial parameters were optimal when cell discharge led the animal's position Furthermore, the optimal time shifts for two of these parameters (firing area and information content) were less than the optimal shift reported for hippocampal place cells and suggested that PaS cell discharge lagged behind hippocampal place cell activity Rotation of the cue card with the animal out of view led to near equal rotation of the firing field when the animal was returned to the apparatus These results indicate that a small population of cells in the PaS encode the animal's location in its environment, although the representation of space encoded by these cells is different from the type of representation encoded by hippocampal place cells

Information acquired by the hippocampus interferes with acquisition of the amygdala-based conditioned-cue preference in the rat.

Abstract: White and McDonald (1993, Behav Brain Res 55:269-281) previously reported that animals with amygdala lesions failed to acquire a conditioned-cue preference (CCP) based on spatial cues, but that animals with fornix lesions exhibited larger CCPs of this type than normal animals. The present experiments focused on the hippocampal interference with amygdala-based CCP learning inferred from this finding. In experiment 1 we tested the hypothesis that this interference was due to the acquisition of information about the maze and its environment during a 10 min period of free exploration of the maze before the start of CCP training, hitherto given to all animals in these experiments. Normal animals that were not preexposed to the maze and animals that were preexposed to a similar maze in a different room both exhibited larger CCPs than animals that were preexposed to the same maze in the same room as CCP training and testing. This suggests that normal animals acquire context-specific information during the preexposure period and that this may be the cause of the hippocampus-based interference with the amygdala-mediated CCP. In experiment 2 we attempted to determine if the information thought to be acquired by the hippocampal memory system interferes with acquisition or expression of the CCP. As previously demonstrated, animals that received fornix lesions before preexposure (i.e., before the start of the experiment) exhibited larger than normal CCPs. Animals that received fornix lesions after preexposure but before CCP training and animals that received fornix lesions after CCP training but before testing both exhibited normal CCPs.(ABSTRACT TRUNCATED AT 250 WORDS)

Simulation of spatial learning in the Morris water maze by a neural network model of the hippocampal formation and nucleus accumbens

Abstract: Cells in the hippocampal formation show spatial firing correlates thought to be critical to the role played by this structure in spatial learning. Place cells in the hippocampus proper show location-specific activity, whereas cells in the postsubiculum fire as a function of momentary directional heading. One question which has received little attention is how these spatial signals are used by motor structures to actually guide spatial behavior. Here we present a model of how one kind of spatial behavior, instrumental learning in the Morris water maze, could be guided by the spatial information in the hippocampal formation. For this, we concentrate on the hippocampal projection to the nucleus accumbens, which is strongly implicated in instrumental learning. In the model, simulated firing patterns of place cells and head direction cells activate "motor" cells in the "accumbens." Each motor cell causes a particular locomotor movement in a simulated rat. In this way, the "rat" locomotes through the simulated environment. Each step places the animal in a slightly different location and directional orientation, which, in turn, activates a different set of place and head direction cells, thus causing the next locomotor response, and so on. Connection strengths between cells are initially set randomly. When the animal encounters the reward location, however, connections are altered, so that recently active synapses are strengthened. Thus, successful moves in a particular locational and directional context are "stamped in." Simulated rats show rapid learning, similar in many ways to that of actual rats. In particular, they generate efficient routes to the goal after minimal experience, and can do so from somewhat novel starting positions. Consideration of the model architecture shows that 1) combined use of directional and place information is an example of a linearly inseparable problem and that 2) some types of novel route generation, often thought to require a "cognitive mapping" strategy, can be generated from the S-R type model used here.

Stable maintenance of glutamate receptors and other synaptic components in long‐term hippocampal slices

Abstract: Cultured hippocampal slices retain many in vivo features with regard to circuitry, synaptic plasticity, and pathological responsiveness, while remaining accessible to a variety of experimental manipulations. The present study used ligand binding, immunostaining, and in situ hybridization assays to determine the stability of AMPA- and NMDA-type glutamate receptors and other synaptic proteins in slice cultures obtained from 11 day postnatal rats and maintained in culture for at least 4 weeks. Binding of the glutamate receptor ligands [3H]AMPA and [3H]MK-801 exhibited a small and transient decrease immediately after slice preparation, but the binding levels recovered by culture day (CD) 5-10 and remained stable for at least 30 days in culture. Autoradiographic analyses with both ligands revealed labeling of dendritic fields similar to adult tissue. In addition, slices at CD 10-20 expressed a low to high affinity [3H]AMPA binding ratio that was comparable with that in the adult hippocampus (10:1). AMPA receptor subunits GluR1 and GluR2/3 and an NMDA receptor subunit (NMDAR1) exhibited similar postcutting decreases as that exhibited by the ligand binding levels, followed by stable recovery. The GluR4 AMPA receptor subunit was not evident during the first 10 CDs but slowly reached detectable levels thereafter in some slices. Immunocytochemistry and in situ hybridization techniques revealed adult-like labeling of subunit proteins in dendritic processes and their mRNAs in neuronal cell body layers. Long-term maintenance was evident for other synapse-related proteins, including synaptophysin, neural cell adhesion molecule isoforms (NCAMs), and an AMPA receptor related antigen (GR53), as well as for certain structural and cytoskeletal components (e.g., myelin basic protein, spectrin, microtubule-associated proteins). In summary, following an initial and brief depression, many synaptic components were expressed at steady-state levels in long-term hippocampal slices, thus allowing the use of such a culture system for investigations into mechanisms of brain synapses.

Contribution of synapses in the medial supramammillary nucleus to the frequency of hippocampal theta rhythm in freely moving rats

Abstract: We have previously shown that in urethane-anesthetized rats the frequncy of rhythmical slow activity in the hippocampus (“theta”) is controlled by the medial supramammillary nucleus (SuM). In particular, injections of procaine into SuM in urethane-anesthetized animals reduce the frequency of theta. However, it has been reported that, in freely moving animals, lesions of SuM do not affect theta. The present experiments were designed to resolve this anomaly. Injections of procaine or chlordiazepoxide into SuM in urethane-anesthetized animals reduced the frequency of theta elicited by reticular stimulation. Mapping showed that procaine injections in freely moving animals were effective in the same locations as under urethane anesthesia. Injections of chlordiazepoxide were effective in a more restricted area than procaine, consistent with an action on synapses in SuM and sparing fibers afferent to SuM. Analysis of the functional spread indicated an effective radius of diffusion of the drugs of 500μm. With optimal placements, this implied an action on at least 80% of SuM. However, in contrast to the results under urethane, the maximal frequency reductions obtained were less than 50% of the theoretical maximum. In a number of animals receiving repeated injections into SuM, lesions developed which encompassed the whole of SuM. As previously reported, theta was largely intact in SuM-lesioned animals. However, the frequency of theta produced by reticular stimulation was reduced after lesion by approximately the same amount as by procaine injections before lesion. These results suggest that in freely moving animals SuM is only one of two or more nuclei which jointly control the frequency of reticular-elicited theta. © 1995 Wiley-Liss, Inc.

Reduction of the threshold for long‐term potentiation by prior theta‐frequency synaptic activity

Abstract: Activation of the lateral perforant path input to the dentate gyrus with theta-patterned conditioning trains produced LTP of synaptic efficacy that changed in magnitude as an inverted U-shaped function of the number of trains. The LTP induction function was not fixed, however, and could be shifted to the left by administering 5 Hz “priming” stimulation to the lateral path 10 min prior to the conditioning trains. The priming effect was input specific and selective to a narrow window of stimulus frequencies. The shift to the left of the LTP induction function by priming stimulation was blocked by the muscarinic receptor antagonist atropine sulphate. Nimodipine, an antagonist of L-type voltage-sensitive calcium channels, did not mimic the priming effect but instead produced a general facilitation of LTP is induction. These data demonstrate that the degree to which LTP is induced in the lateral path is a non-linear function of afferent activity, and that this function, including LTP threshold, can be shifted to the left by prior synaptic activity at hippocampal theta-rhythm frequencies. © 1995 Wiley-Liss, Inc.

Postischemic inhibition of GABA reuptake by tiagabine slows neuronal death in the gerbil hippocampus.

Abstract: The neuroprotective effects of enhancing neuronal inhibition with a γ-aminobutyric acid (GABA) uptake inhibitor were studied in gerbil hippocampus following transient ischemia. We used in vivo microdialysis to determine a suitable dosing regimen for tiagabine (NNC 328) to elevate extracellular levels of GABA within the hippocampus. In anesthetized (normothermic) gerbils, tiagabine (45 mg/kg, i. p.) selectively elevated extracellular GABA levels 450% in area CA1 of the hippocampus. In gerbils subjected to cerebral ischemia via 5-min bilateral carotid occlusion, extracellular GABA levels increased 13-fold in area CA1, returning to baseline within 30–45 min. When tiagabine was injected 10 min following onset of reperfusion, GABA levels remained elevated (200–470%) for 90 min. In addition, tiagabine significantly reduced the ischemic-induced elevation of glutamate levels in area CA1 during the postischemic period when GABA levels were elevated. There was no effect of postischemic tiagabine on aspartate or six other amino acids. Using the same dosing regimen, we evaluated the degree of neuroprotection in the hippocampus of gerbils 4 and 21 days after ischemia. Tiagabine decreased body temperature a maximum of 2.7°C beginning 30 min into reperfusion and lasting 90 min. In untreated gerbils sacrificed 4 and 21 days after ischemia, there was severe necrosis (99%) of the pyramidal cell layer in area CA1. Whereas tiagabine significantly protected the CA1 pyramidal cell layer in ischemic gerbils at 4 days (overt necrosis confined to about 17% of area CA1), the protection diminished significantly 21 days postischemia. When normothermia was maintained both during and after ischemia in a separate group of tiagabine-treated animals, approximately 77% of the CA1 pyramidal cell layer was necrotic at 4 days. Based on these findings, we suggest that (1) tiagabine slows the development of hippocampal degeneration following ischemia, and (2) that mild, postischemic hypothermia is responsible, in large part, for the neuroprotective actions of this drug. We conclude that the histological outcome after administration of cerebral neuroprotectants should be assessed following long-term survival. © 1995 Wiley-Liss, Inc.

Dentate gyrus destruction and spatial learning impairment after corticosteroid removal in young and middle-aged rats.

Abstract: We investigated the functional and behavioral implications of chronic corticosteroid removal in young and middle-aged rats. Prepubertal and 13-month-old rats were adrenalectomized (ADX) or sham operated (SHAM). The young ADX rats were divided further into three groups: ADX with no hormone replacement, ADX given corticosterone chronically, (chCORT), and ADX given corticosterone acutely at the time of Morris water maze testing (acCORT). All rats were run on the Morris water maze 12 weeks after surgery. They were then sacrificed and the brains were removed for histological analysis. The results showed that prolonged corticosteroid absence caused ma- jor damage to the dentate gyrus and learning impairment on the Morris water maze. The chCORT rats had little dentate gyrus cell loss and were as efficient as the controls in Morris water maze performance, whereas the acCORT rats had dentate gyrus cell loss and were impaired in the spatial acquisition task. Furthermore, exogenously administered corti- costerone had an interactive effect on ADX rats. Water maze performance was improved in dentate gyrus damaged rats (acCORT) compared to ADX rats not given corticosterone, whereas ADX rats with very little dentate gyrus damage (chCORT) did not exhibit better water maze performance relative to controls. Middle-aged ADX rats lost cells only in the dorsal blade of the dentate gyrus but they did not show a learning impairment in the Morris water maze relative to the middle-aged controls. These re- sults indicate that corticosteroids are trophic for the dentate gyrus, that mature granule cells are less affected by adrenalectomy, that cortico- steroid absence is responsible for some water maze impairment in ADX rats, but that in addition to corticosteroid absence, a substantial amount of dentate gyrus damage is necessary to impair spatial learning. D 1995 Wiley-Liss, Inc.

Classification of theta-related cells in the entorhinal cortex: cell discharges are controlled by the ascending brainstem synchronizing pathway in parallel with hippocampal theta-related cells.

Abstract: Single-unit discharge patterns of entorhinal cortex (EC) cells were characterized in relation to simultaneously recorded hippocampal (HPC) field activity according to criteria used previously to classify cells in the hippocampal formation, medial septum, cingulate cortex, and caudal diencephalon. EC cells related to HPC theta field activity were classified as (1) phasic theta-on, if they discharged rhythmically, and in phase, with ongoing HPC theta, but nonrhythmically during large, irregular hippocampal field activity (LIA); (2) tonic theta-on, if they discharged nonrhythmically and increased their discharge rates during HPC theta relative to LIA; (3) phasic theta-off, if they discharged rhythmically, and in phase, with ongoing HPC theta, but increased their discharge rates during LIA; and (4) tonic theta-off, if they discharged nonrhythmically and decreased their discharge rates during HPC theta relative to LIA. Cells not meeting any of these criteria were classified as nonrelated. A total of 168 EC cells were recorded, and of these 56 (33%) were classified as theta related, with the remaining 112 (67%) classified as nonrelated. Of the 56 theta-related cells, 41 (73%) had significantly higher discharge rates during HPC theta than during LIA and were classified as theta-on cells (15 phasic theta-on cells and 26 tonic theta-on cells). Nine of the 26 tonic theta-on cells showed a phase relation of their arrhythmic discharges to simultaneously recorded HPC theta field activity. EC phasic theta-on cells did not discharge preferentially on any portion of the HPC theta field recorded from the region of the stratum moleculare of the dentate gyrus. In general, cells classified as phasic revealed a wide distribution of phase preferences. The remaining 15 (26.7%) cells were classified as theta-off cells and discharged at higher rates during HPC LIA than during HPC theta field activity (3 phasic theta-off cells and 12 tonic theta-off cells). Systemic administration of physostigmine significantly increased the discharge rate of tonic and phasic theta-on cells relative to LIA. Electrical stimulation in the posterior hypothalamic region (PH) significantly increased the discharge rate of EC theta-on cells and significantly decreased the discharge rate of EC theta-off cells relative to HPC LIA. The discharge rates of nonrelated EC cells were not influenced by electrical stimulation of the PH. Procaine microinfusion into the medial septum (MS) abolished spontaneously occurring HPC theta and theta induced with PH stimulation. In addition, 5 min after MS procaine, the ability of PH stimulation to modulate EC theta-on cell discharge was abolished. The modulation of cellular discharges produced by PH stimulation recovered by 60 min post-procaine infusion into the MS. The findings support two main conclusions: (1) theta-related cells in the EC are comprised of two main populations of cells, theta-on and theta-off, similar to other regions of limbic cortex and nuclei of the ascending brainstem synchronizing pathway; (2) the ascending brainstem synchronizing pathway exerts both similar and parallel effects on theta-related cells in entorhinal cortex and hippocampus. © 1995 Wiley-Liss, Inc.

Electrophysiological Diversity of Pyramidal-Shaped Neurons at the Granule Cell Layer/Hilus Border of the Rat Dentate Gyrus Recorded In Vitro

Abstract: In the rat dentate gyrus, pyramidal-shaped cells located on the border of the granule cell layer and the hilus are one of the most common types of gamma-aminobutyric acid (GABA)-immunoreactive neurons. This study describes their electrophysiological characteristics. Membrane properties, patterns of discharge, and synaptic responses were recorded intracellularly from these cells in hippocampal slices. Each cell was identified as pyramidal-shaped by injecting the marker Neurobiotin intracellularly (n = 17). In several respects the membrane properties of the sampled cells were similar to "fast-spiking" cells (putative inhibitory interneurons) that have been described in other areas of the hippocampus. For example, input resistance was high (mean 91.3 megohms), the membrane time constant was short (mean 7.7 ms), and there was a large afterhyperpolarization following a single action potential (mean 10.5 mV at resting potential). However, the action potentials of most pyramidal-shaped cells were not as brief (mean 1.2 ms total duration) as those of most previously described fast-spiking cells. Many pyramidal-shaped neurons had strong spike frequency adaptation relative to other fast-spiking cells. Although these latter two characteristics were apparent in the majority of the sampled cells, there were exceptional pyramidal-shaped neurons with fast action potentials and weak adaptation, demonstrating the electrophysiological variability of pyramidal-shaped cells. Responses to outer molecular layer stimulation were composed primarily of excitatory postsynaptic potentials (EPSPs) rather than inhibitory postsynaptic potentials (IPSPs), and were usually small (EPSPs evoked at threshold were often less than 2 mV), and brief (less than 30 ms). There was variability, because in a few cells EPSPs evoked at threshold were much larger. However, regardless of EPSP amplitude, suprathreshold stimulation (up to 4 times the threshold stimulus strength) rarely evoked more than one action potential in any cell. The results suggest that stimulation of perforant path axons produces limited excitatory synaptic responses in pyramidal-shaped neurons. This may be one of the reasons why they are relatively resistant to prolonged perforant path stimulation. The pyramidal-shaped neurons located at the base of the granule cell layer have been associated historically with a basket plexus around granule cell somata, and have been called pyramidal "basket" cells. However, basket-like endings were rare and axon collaterals outside the granule cell layer as the outer molecular layer and the central hilus, and antidromic action potentials could be recorded in some cells in response to weak stimulation of these areas. Taken together with the electrophysiological variability, the results indicate that these cells are physiologically heterogeneous.

Postnatal development of CA3 pyramidal neurons and their afferents in the Ammon's horn of rhesus monkeys.

Abstract: Previous studies described the postnatal development of CA3 pyramidal neurons and their afferents in the rat. However, the postnatal development of the primate hippocampus was not previously studied. Thus, pyramidal neurons of the CA3 area of the monkey hippocampus were analyzed postnatally in the present study. At birth, a few thorny excrescences, the complex spines postsynaptic to mossy fibers, were found on the proximal segments of both apical and basal dendrites, whereas distal dendrites displayed pedunculate spines. Thorny excrescences increased in number until the third month. A continuous increase in the number of spines per unit length along the distal dendrites was observed during the first 12 months. The ultrastructural features of somata and dendrites of pyramidal cells in newborn monkeys were similar to those of adults. The analysis of the afferents to the CA3 pyramidal neurons was limited to the development of mossy fibers, the axons of granule cells, and myelinated axons in the alveus, stratum oriens, and stratum lacunosum-moleculare. At birth, most mossy fiber terminals were densely packed with synaptic vesicles and formed mainly axospinous synapses with CA3 pyramidal cells. By 1 month of age, the number of mitochondria and embedded spines increased to mature amounts. In the first postnatal month, degenerating axons and axon terminals were frequently observed in the mossy fiber bundles in stratum lucidum. The proportion of myelinated axons increased simultaneously in all three examined layers. At birth most axons were unmyelinated, whereas at 7 months of age the proportion of myelinated axons was similar to that found in adults. The present study indicates that most pyramidal neurons of the CA3 region in monkeys are in an advanced stage of development at the time of birth. Thus, mossy fibers from granule cells in the dentate gyrus have established mature-looking synapses, and the thorny excrescences of pyramidal cells that are postsynaptic to mossy fibers are also adult-like. Nevertheless, several of the adult features, such as the spine density of distal dendrites of pyramidal neurons and the myelination of afferent axons, develop during an extended period of time in the first year. The significance of this early anatomical maturation in a brain region involved in memory function is consistent with recent behavioral data that show a rapid postnatal maturation of limbic-dependent recognition memory in rhesus monkeys.

Alterations in the immunoreactivity for muscarinic acetylcholine receptors and colocalized PKCγ in mouse hippocampus induced by spatial discrimination learning

Abstract: This study describes changes in the immunoreactivity for muscarinic acetylcholine receptors (mAChRs) in the hippocampus of mice in relation to spatial discrimination behavior, employing the monoclonal antibody M35 raised against purified bovine mAChR protein. Performance in a hole board in which the animals learned the pattern of 4 baited holes out of 16 holes served as the measure of spatial discrimination learning and memory. Twenty-six adult male house mice were used, divided into four groups. Three groups served as various controls: group N (naive; blank controls); group H (habituated; animals were introduced to the hole board with all holes baited for 5 consecutive days), and group P (pseudo-trained; the animals were admitted to the hole board for 13 consecutive days with all holes baited). The T group (trained) was subjected to the hole board for 5 consecutive habituation days with all holes baited (similar to the H and P groups), followed by 8 successive training days with only four holes baited in a fixed pattern. During the 8 training days, the T group gradually acquired a pattern to visit the baited holes, whereas the P group continued visiting holes in a random fashion. The mice were killed 24 h after the last behavioral session. All principal cells in teh cornu ammonis (CA) and dentate gyrus (DG) of the habituated animals revealed increased levels of mAChR immunoreactivity (mAChR-ir) over the naive mice. A minor increase in mAChR-ir was found in the apical dendrites of the CA1 pyramidal cells. Pseudotraining resulted in a CA1-CA2 region with a low level of mAChR-ir, resembling naive animals, whereas the trained mice showed a further increase in mAChR-ir in the CA1-CA2 pyramidal cell bodies and apical dendrites. Optical density measures of the mAChR-ir in the CA1 region revealed a significant (P < 0.05) increase in the pyramidal cell bodies of the H and T group over the N and P group, and a significant (P < 0.05) increase in the apical dendrites of the T group over all other groups. In contrast to the CA1-CA2 region, both pseudotrained and trained mice revealed high mAChR staining in the CA3-CA4 region and the DG. These results indicate that prolonged exposure to the hole board is sufficient for an enhanced mAChR-ir in the CA3-CA4 and DG, whereas the increase in CA1-CA2 pyramidal cells is a training-specific feature related to spatial orientation. Nonpyramidal neurons within the CA1-CA2 region with enhanced mAChR-ir in the pyramidal cells, however, revealed a decreased level of mAChR-ir. The opposing effect of pyramidal and nonpyramidal cells suggests a shift in the excitability of the hippocampal microcircuitry. Previously we demonstrated an increase and redistribution of hippocampal protein kinase C gamma-immunoreactivity (PKC gamma-ir) induced by hole board learning in mice (Van der Zee et al., 1992, J Neurosci 12:4808-4815). Immunofluorescence double-labeling experiments conducted in the present study in naive and trained animals revealed that the principal cells and DG interneurons co-express mAChRs and PKC gamma, and that the immunoreactivity for both markers increased in relation to spatial orientation within these neurons. The mAChR-positive nonpyramidal cells of the CA1-CA2 region were devoid of PKC gamma and revealed an opposite training-induced effect. These results suggest that the postsynaptic changes in mAChR- and PKC gamma-ir reflect functional alterations of the hippocampal formation induced by spatial learning.

Spontaneous and synaptic input from granule cells and the perforant path to dentate basket cells in the rat hippocampus.

Abstract: To characterize excitatory inputs to dentate basket cells from dentate granule cells and the perforant path, the whole-cell recording technique was used in neonatal rat hippocampal slices Spontaneous excitatory input to basket cells was also examined and compared to that of other interneurons in the dentate gyrus Basket cells were separable from other neurons in the dentate gyrus based on morphology and location, as determined by biocytin staining following recording, and by resting membrane potential, propensity to fire action potentials spontaneously, and miniature excitatory postsynaptic current (EPSC) characteristics Minimal electrical stimulation of the granule cell layer evoked in basket cells short latency EPSCs that were composed of both N-methyl-D-aspartate (NMDA) and α-amino-3-hydroxy-5-methyl-4-isoxazole-propionate (AMPA) components as judged by their time course, voltage dependence, and blockade by selective antagonists Perforant path EPSCs exhibited slower kinetics than EPSCs evoked by granule cell stimulation Like granule cell evoked EPSCs, however, perforant path EPSCs were composed of both NMDA and AMPA components Minimal electrical stimulation of the granule cell layer and perforant path evoked monosynaptic EPSCs in only 67% and 62% of the trials, respectively, suggesting that these inputs are as unreliable as previously determined inputs from CA3 pyramidal cells (48%) Tetrodotoxin-insensitive spontaneous miniature EPSCs were frequent in basket cells and non-basket interneurons residing either at the border between the granule cell layer and the hilus or deep within the hilus Miniature EPSCs recorded from all cells held at −70 mV were blocked completely by 3 μSM 6-cyano-7-nitro-quinoxaline-2,3-dione (CNQX) Though a component of the miniature EPSCs recorded from border and deep hilar interneurons at +40 mV was blocked by the NMDA receptor antagonist D-2-amino-phosphonovaleric acid (D-APV), miniature EPSCs in basket cells were insensitive to D-APV We conclude that input from granule cells and the perforant path results in activation of basket cells via glutamatergic synapses that employ both NMDA and AMPA receptors These inputs to basket cells likely contribute to feedback and feedforward inhibition of granule cells The absence of an NMDA receptor component in spontaneous miniature EPSCs of dentate basket cells implies a difference in organization of excitatory synapses made onto basket cells compared with other hilar interneurons © 1995 Wiley-Liss, Inc

Changes in exploratory activity following stimulation of hippocampal 5‐HT1A and 5‐HT1B receptors in the rat

Abstract: The object exploration task allows the measure of changes in locomotor and exploratory activities, habituation, and reaction to a spatial change and to novelty. The effects of intrahippocampal (dorsal CA1 field) microinjections of serotonin 1 receptor (5-HT1) agonists on these behavioral components were evaluated in the rat. 8-Hydroxy-2-(di-n-propylamino)-tetralin (8-OH-DPAT, 5 micrograms/microliters) was used as a 5-HT1A agonist, 3-(1,2,5,6-tetrahydropyrid-4-yl)pyrrolo[3,2-b]pyrid-5-one (CP 93,129,16 micrograms/microliters) as a 5-HT1B agonist, and scopolamine (10 micrograms/microliters) as a muscarinic cholinergic antagonist. Scopolamine induced a long-lasting increase in locomotor activity and a lack of reaction to spatial change; both these results are in agreement with the known crucial influence of the septo-hippocampal cholinergic system in hippocampal functioning. Stimulation of 5-HT1A and 5-HT1B receptors induced a decrease in object exploration and habituation without affecting the retrieval of spatial information. But stimulation of hippocampal 5-HT1B receptors induced a selective change in the animal's emotional state, i.e., an initial decrease in locomotor activity and a neophobic reaction in response to a new object; such effects did not occur following stimulation of 5HT1A receptors. These results have to be considered in the light of the anxiogenic property of 5-HT1B agonists. On the whole, they support the hypothesis of the involvement of the serotonergic system, via 5HT1A and 5-HT1B receptors, in the modulation of hippocampal functions.

AMPA-selective glutamate receptor subtype immunoreactivity in the hippocampal formation of patients with Alzheimer's disease

Abstract: Immunocytochemical techniques were employed in order to examine the distribution and relative intensity of the AMPA receptor subunits GluR1 and GluR2/3 within the hippocampal formation of normal controls and Alzheimer's disease (AD) cases. Throughout our investigation we examined cases exhibiting a wide range of pathologic severity, thus allowing us to correlate our immunohistochemical data with the extent of pathology. Specifically, we investigated the distribution of these receptor subunits in hippocampal sectors that are particularly vulnerable to AD pathology (i.e., CA1 and subiculum) and compared these findings with those obtained following examination of sectors that are generally resistant to pathologic change (i.e., CA2/3, dentate gyrus). Within vulnerable sectors we observed a variable loss of GluR1 and GluR2/3 immunolabeling. The degree to which these proteins were reduced appeared to correlate with the extent of neurofibrillary pathology and cell loss. Despite the loss of labeled cells, the intensity of immunolabeling within the remaining neurons was comparable with, and in many instances even greater than, that observed in control cases. Within resistant sectors, the distribution of immunoreactive elements was comparable in both case groups yet the intensity of immunolabeling was markedly increased in AD cases, particularly in the molecular layer of the dentate gyrus and in the stratum lucidum of CA3 (i.e., the termination zones of perforant pathway and mossy fibers). In addition, within AD cases dramatic increases were observed within the supragranular and polymorphic layer of the dentate gyrus (i.e., the terminal zones of sprouting mossy fiber collaterals). The increase in GluR1 and GluR2/3 immunolabeling is hypothesized to occur in response to the deafferentation of selected glutamatergic pathways. Moreover, our data support that hippocampal plasticity is preserved, even in severe AD cases, and suggest a critical role for AMPA receptor subunits in this plasticity and in maintaining hippocampal functioning.

Adenosine antagonists have differential effects on induction of long-term potentiation in hippocampal slices.

Abstract: How adenosine leakage and tetanic release might affect long-term potentiation (LTP) was investigated by applying adenosine an- tagonists 8(p-sulfophenyl)theophylline (8SPT) or 8-cyclopentyl-3,7-dihy- dro-l,3-dipropyl-l H-purine-2,6-dione (DPCPX) to slices, while recording CA1 field EPSPs and population spikes. In the first series of experiments, we applied weak double tetani (at 100 Hz, for 1 s) that were subliminal for evoking LTP in initial control runs. In the presence of 8SPT-at con- centrations (10-50 pM) which block both A, and A2 receptors-the same tetani consistently evoked LTP of population spikes but not of excitatory postsynaptic potentials (EPSPs), whereas DPCPX (50 nM), which blocks only Al receptors, facilitated LTP of both EPSPs and population spikes. These results are consistent with previous evidence that tetanic adeno- sine release on the one hand depresses LTP via A, receptors but on the other facilitates LTP via AZ receptors. In a second set of experiments, 8SPT (50-1 00 pM) did not prevent the induction of LTP of both EPSPs and population spikes by stronger tetanic stimulation. Therefore A2 receptor activation is not essential for the in- duction of LTP when stronger tetani are applied. Overall, the main effect of endogenous adenosine release is to oppose LTP induction. 0 1995 Wiley-Liss, Inc.