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Sequential involvement of NMDA receptor-dependent processes in hippocampus, amygdala, entorhinal cortex and parietal cortex in memory processing.

Rats bilaterally implanted with cannulae in the CA1 region of the dorsal hippocampus and/or in the amygdaloid nucleus, in the entorhinal cortex, and in the posterior parietal cortex, were trained in a step-down inhibitory avoidance task. At various times after training (immediately, 30, 60 or 90min) they received, through the cannulae, 0.5µl microinfusions of saline or of 5.0µg of AP5 dissolved in saline. A retention test was carried out 24h after training. Retention test performance was hindered by AP5 given into hippocampus, amygdala, or both hippocampus and amygdala immediately but not 30min post-training. The drug was amnestic when given into the entorhinal cortex 30, 60 or 90min after training, or into the parietal cortex 60 or 90min after training, but not at earlier times. The findings suggest a sequential entry in operation, in the post-training period, of NMDA-receptor mediated mechanisms involved in memory processing; first in hippocampus and amygdala, 30min later in entorhinal cortex, and 30min later in posterior parietal cortex.

Journal Article↗

Behavioral correlates of denervation and reinnervation of the hippocampal formation of the rat: open field activity and cue utilization following bilateral entorhinal cortex lesions.

Bilateral lesions of the entorhinal cortex (E.C.) of the rat result in persistent deficits in both spontaneous and reinforced alternation. The present study analyzes the nature of this impairment. To determine if changes in exploratory activity accompanied the deficits in alternation, open field activity was measured daily from 2-22 days following bilateral E.C. lesions. Such lesions resulted in a pronounced transient increase in open field activity which peaked between 5 and 7 days postlesion, but subsequently decreased to near preoperative levels at approximately 11 days postlesion. Alternation performance was also analyzed, to determine which cues are utilized to make the alternation, and whether cue utilization is affected by bilateral E.C. lesions. Utilizing a plus (+) maze, animals readily learned to alternate goal arms, but even with extensive training, failed to learn to alternate turns (left and right). However, the ability to identify the two goal arms in a nonalternation situation (which does not require short term recall of the preceding trial) was not permanently impaired by bilateral E.C. lesions. Since bilateral E.C. lesions do not result in persistent deficits in the ability to identify the two goal arms, but do disrupt alternation performance, we hypothesize that the deficit in alternation might reflect an inability to recall which arm was chosen on preceding trials. The implications of these results for an understanding of the behavioral consequences of postlesion reorganization of neuronal circuitry are discussed.

Animals↗

Gamma oscillations in the entorhinal cortex of the freely behaving rat.

Gamma frequency field oscillations (40-100 Hz) are nested within theta oscillations in the dentate-hilar and CA1-CA3 regions of the hippocampus during exploratory behaviors. These oscillations reflect synchronized synaptic potentials that entrain the discharge of neuronal populations within the approximately 10-25 msec range. Using multisite recordings in freely behaving rats, we examined gamma oscillations within the superficial layers (I-III) of the entorhinal cortex. These oscillations increased in amplitude and regularity in association with entorhinal theta waves. Gamma waves showed an amplitude minimum and reversed in phase near the perisomatic region of layer II, indicating that they represent synchronized synaptic potentials impinging on layer II-III neurons. Theta and gamma oscillations in the entorhinal cortex were coupled with theta and gamma oscillations in the dentate hilar region. The majority of layer II-III neurons discharged irregularly but were phase-related to the negative peak of the local (layer II-III) gamma field oscillation. These findings demonstrate that layer II-III neurons discharge in temporally defined gamma windows (approximately 10-25 msec) coupled to the theta cycle. This transient temporal framework, which emerges in both the entorhinal cortex and the hippocampus, may allow spatially distributed subpopulations to form temporally defined ensembles. We speculate that the theta-gamma pattern in the discharge of these neurons is essential for effective neuronal communication and synaptic plasticity in the perforant pathway.

Animals↗

Lesion-induced plasticity of central neurons: sprouting of single fibres in the rat hippocampus after unilateral entorhinal cortex lesion.

In response to a central nervous system trauma surviving neurons reorganize their connections and form new synapses that replace those lost by the lesion. A well established in vivo system for the analysis of this lesion-induced plasticity is the reorganization of the fascia dentata following unilateral entorhinal cortex lesions in rats. After general considerations of neuronal reorganization following a central nervous system trauma, this review focuses on the sprouting of single fibres in the rat hippocampus after entorhinal lesion and the molecular factors which may regulate this process. First, the connectivity of the fascia dentata in control animals is reviewed and previously unknown commissural fibers to the outer molecular layer and entorhinal fibres to the inner molecular layer are characterized. Second, sprouting of commissural and crossed entorhinal fibres after entorhinal cortex lesion is described. Single fibres sprout by forming additional collaterals, axonal extensions, boutons, and tangle-like axon formations. It is pointed out that the sprouting after entorhinal lesion mainly involves unlesioned fibre systems terminating within the layer of fibre degeneration and is therefore layer-specific. Third, molecular changes associated with axonal growth and synapse formation are considered. In this context, the role of adhesion molecules, glial cells, and neurotrophic factors for the sprouting process are discussed. Finally, an involvement of sprouting processes in the formation of neuritic plaques in Alzheimer's disease is reviewed and discussed with regard to the axonal tangle-like formations observed after entorhinal cortex lesion.

Animals↗

Autoradiographic localization of estradiol-binding neurons in the rat hippocampal formation and entorhinal cortex.

This study has examined the distribution of [3H]estradiol and [1 alpha,2 alpha-3H]testosterone uptake in the hippocampal formation and entorhinal cortex of male and female rats. In both males and females, [3H]estradiol-binding neurons in Ammon's horn are located deep in stratum pyramidale and may correspond either to polymorphic interneurons or to early maturing pyramidal cells. Interneurons of strata oriens, lucidum and radiatum of Ammon's horn and of stratum moleculare of the subiculum also bind [3H]estradiol, as do basket cell interneurons in the polymorphic, infragranular layer of the dentate gyrus. While no granule cells appear to accumulate [3H]estradiol, these cells may be affected transsynaptically by gonadal steroids via their afferent contacts with the entorhinal cortex, which, of the areas examined, contains the greatest number of [3H]estradiol-binding neurons. While relatively few neurons concentrate [3H]estradiol in the hippocampal formation, these are localized to specific subpopulations, which may enhance their functional significance. Because there is no significant nuclear accumulation of [3H]-alpha-testosterone in either the entorhinal cortex or hippocampal formation, it appears that aromatase enzyme activity is not a major contributor to estrogen receptor occupancy in adult rats.

Animals↗

Localization of cytochrome oxidase (COX) activity and COX mRNA in the hippocampus and entorhinal cortex of the monkey brain: correlation with specific neuronal pathways.

Cytochrome oxidase (COX) activity and COX II mRNA expression were localized in the hippocampal formation and entorhinal cortex of the rhesus monkey brain by means of enzyme histochemistry and in situ hybridization, respectively. Within the hippocampal formation, the terminal field of the perforant pathway showed the highest levels of COX activity, whereas COX II mRNA was localized mainly in neuronal cell bodies. In the entorhinal cortex. COX II mRNA was detected in neuronal cell bodies of layers II and IV. These results indicate that the pattern of localization of COX and its mRNA in entorhinal cortex correlates with the input and output pathways of the hippocampus.

Animals↗

Stereologic analysis of hippocampal Alzheimer's disease pathology in the oldest-old: evidence for sparing of the entorhinal cortex and CA1 field.

Several neuropathologic analyses postulate that Alzheimer disease (AD) in the oldest-old is associated with substantial neurofibrillary tangle (NFT) formation in the CA fields of the hippocampus and neuronal loss confined to the entorhinal cortex. All of these studies have measured densities, rather than absolute numbers, and most do not take into account the potential interaction between the above pathological hallmarks in a global multivariate analysis. We present here a stereologic analysis of AD-related pathology in 12 oldest-old individuals including a complete assessment of total NFT, neuron numbers and amyloid volume in entorhinal cortex, CA fields, and dentate gyrus. The progression of NFT numbers and amyloid volume across the different Clinical Dementia Rating (CDR) groups was significantly slower in these cases compared to previously reported younger cases. Although patients with mild and moderate dementia showed significantly lower mean neuron numbers compared to CDR 0-0.5 cases, there was a marked overlap in individual values among CDR groups. A modest proportion of the variability in CDR scores was explained by NFT numbers in the CA2 field (18.1%) and the dentate gyrus (17.3%). In contrast, neither Nissl-stained neuron numbers nor total amyloid volume in the areas studied significantly predicted cognitive status. These data indicate that the occurrence and progression of AD-related pathologic changes are not an unavoidable consequence of aging. They also suggest that dementia in extreme aging depends more on the damage of hippocampal subdivisions commonly less affected than on severe NFT formation and neuronal loss in the CA1 field and entorhinal cortex.

Aged↗

Late low magnesium-induced epileptiform activity in rat entorhinal cortex slices becomes insensitive to the anticonvulsant valproic acid.

We investigated time-dependent changes in low magnesium-induced epileptiform activity in combined rat entorhinal cortex/hippocampal slices with extracellular recording techniques. While in area CA3 short interictal discharges are generated without any major changes in activity during prolonged recording periods, initial tonic clonic ictaform events in the entorhinal cortex may change with time. We observed often a transition into a state of recurrent tonic activity without any clonic afterdischarges. Alternatively, seizures could stay in the clonic discharge mode for the rest of the experiment. These different seizure states were not equally affected by the anticonvulsant valproic acid. While the early clonic tonic discharges in the entorhinal cortex and the interictal like activity in area CA3 were effectively suppressed by valproic acid (VPA) the late recurrent tonic seizure discharge state was unaffected by the drug. It was, however, still sensitive to the N-methyl-D-aspartate (NMDA) receptor antagonist 2-aminophosphonovalerate. These findings point to seizure-induced changes in neuronal interaction in rat entorhinal cortex.

2-Amino-5-phosphonovalerate↗

Effects of losigamone on synaptic potentials and spike frequency habituation in rat entorhinal cortex and hippocampal CA1 neurones.

Losigamone is an anticonvulsant both in vivo and in vitro. We here studied possible mechanisms for such effects with conventional intracellular recordings from pyramidal cells of area CA1 and entorhinal cortex in combined hippocampal-entorhinal cortex slices. Losigamone reversibly reduced the number of action potentials elicited by 1 s long depolarising current injections. In addition, the drug moderately reduced EPSP amplitudes while monosynaptic fast and slow IPSPs were unaffected.

Action Potentials↗

Precocious development of parvalbumin-like immunoreactive interneurons in the hippocampal formation and entorhinal cortex of the fetal cynomolgus monkey.

The calcium-binding protein parvalbumin (PV), a reliable marker of the hippocampal basket and chandelier cells, is first expressed on embryonic day 83 (E83), corresponding to midgestation of the macaque monkey, in restricted hippocampal groups of immature neurons (Berger and Alvarez [1996] J. Comp. Neurol. 366:674-699). In the present study, PV-like immunoreactivity (LIR) was used to follow the further development of this subclass of interneurons. Asynchronous area-specific developmental sequences were observed, predominating initially in the caudal half of the hippocampal formation and the laterocaudal division of the entorhinal cortex and occurring relatively simultaneously in the interconnected hippocampal and entorhinal subfields. Dendritic elongation of PV-like immunoreactive interneurons and perisomatic distribution of PV-like immunoreactive terminal boutons on their cellular targets were first observed in the subiculum around E127; then from E127 to E142 in CA3/CA2 and layers III-V of the entorhinal cortex and, to a lesser extent in CA1, the dentate hilus and deep granule cell layer; and finally from E156 to postnatal day 12 in the rest of the dentate gyrus, the presubiculum and parasubiculum, and layers III-II-I of the entorhinal cortex. These data provide the first indication that a population of basket cells, a major gamma-aminobutyric acid (GABA)ergic component of the hippocampal intrinsic inhibitory circuitry, reaches its cellular targets several weeks before birth in primates in contrast to rodents. The role of the prenatal PV expression in the hippocampal formation of nonhuman primates and whether it coincides with the onset of postsynaptic inhibitory potentials or is accompanied or preceded by a period of gamma-aminobutyric acid-mediated excitatory effects as in rat pups, are crucial questions. They underline the need to pursue direct investigations on primates to be able to legitimately extrapolate the data obtained in rodents.

Aging↗

Limited responses of neuronal mRNAs to unilateral lesions of the rat entorhinal cortex.

Following unilateral electrolytic lesioning of the rat entorhinal cortex, we assessed changes in messenger RNA (mRNA) levels for four neuron-associated proteins, GAP-43, SCG10, 68 kDa neurofilament (NF68), and alpha 1-tubulin that encode proteins of importance to synaptic remodelling. The mRNA levels for GAP-43 and SCG10 were reduced in the septal nuclei, while those of SCG10 and alpha 1-tubulin were elevated in the hippocampus; at most, the changes ranged from -40% to +50% of controls. Changes in NF68 mRNA levels were not significant. Correlations were found between mRNA for SCG10 and both GAP-43 and NF68. In view of these modest changes in most mRNA levels examined, we suggest that post-translational regulation may also be important in responses to injury.

Animals↗

Divergent cortical connections to entorhinal cortex from area TF in the macaque.

The entorhinal cortex (EC) is an important component of the medial temporal lobe memory system in the primate and is often viewed as a "gatekeeper" area that passes on highly convergent cortical inputs toward the hippocampus. Further analysis of these connections at a microcircuitry level regarding the actual size and shape of arbors and terminations is not yet available, but may contribute to understanding the role of the EC in memory or other functions. The main emphasis of this report was on serial section analysis of anterogradely labeled axons that project from area TF (lateral parahippocampal cortex; Bonin and Bailey, 1947) to the EC (n = 12). By way of evaluating network organization, other projections from area TF--to TH (in the medial parahippocampal gyrus; n = 5) and to posterior visual areas (n = 3)--were also investigated. All three systems were found to terminate heavily in layer 1, as expected from previous investigations, but some terminations were verified in layer 6 of the EC as well. This technique further demonstrated that terminal fields are widely divergent and elongated. In the EC, terminal fields extended over 6-11 mm and spanned multiple cell islands and interislands. These axons resemble "feedback" cortical connections by virtue of their layer 1 terminations and their markedly divergent geometry, but not by their origin from layer 3. Spatially extended terminal fields recall the nontopographic, distributed character of olfactory connections and raise questions of how these features might be related to the memory functions attributed to medial temporal regions.

Animals↗

Molecular and functional analysis of hyperpolarization-activated pacemaker channels in the hippocampus after entorhinal cortex lesion.

Differential display of hippocampal tissue after entorhinal cortex lesion (ECL) revealed decreases in mRNA encoding the neuronal hyperpolarization-activated, cyclic nucleotide-gated channel HCN1. In situ hybridization confirmed that hippocampal transcripts of HCN1, but not HCN2/3/4, are down-regulated after ECL. Expression recovered at approximately 21 days after lesion (dal). Immunohistochemistry demonstrated a corresponding regulation of HCN1 protein expression in CA1-CA3 dendrites, hilar mossy cells and interneurons, and granule cells. Patch-clamp recordings in the early phase after lesion from mossy cells and hilar interneurons revealed an increase in the fast time constant of current activation and a profound negative shift in voltage activation of Ih. Whereas current activation recovered at 30 dal, the voltage activation remained hyperpolarized in mossy cells and hilar interneurons. Granule cells, however, were devoid of any detectable somatic Ih currents. Hence, denervation of the hippocampus decreases HCN1 and concomitantly the Ih activity in hilar neurons, and the recovery of h-current activation kinetics occurs parallel to postlesion sprouting.

Animals↗

Hippocampus-mediated activation of superficial and deep layer neurons in the medial entorhinal cortex of the isolated guinea pig brain.

The entorhinal cortex (EC) is regarded as the structure that regulates information flow to and from the hippocampus. It is commonly assumed that superficial and deep EC neurons project to and receive from the hippocampal formation, respectively. Anatomical evidences suggest that both the hippocampal output and deep EC neurons also project to superficial EC layers. To functionally characterize the interlaminar synaptic EC circuit entrained the by hippocampal output, we performed simultaneous intracellular recordings and laminar profile analysis in the medial EC (m-EC) of the in vitro isolated guinea pig brain after polysynaptic hippocampal activation by lateral olfactory tract (LOT) stimulation. Optical imaging of voltage-generated signals confirmed that the LOT-evoked hippocampus-mediated response is restricted to the m-EC. The hippocampal output generated an extracellular current sink in layers V-VI, coupled with an EPSP in deep neurons. Deep neuron firing was terminated by a biphasic IPSP. The earliest response observed in superficial layer neurons was characterized by a feedforward IPSP of circa 100 ms (-69 +/- 1.3 mV reversal potential) abolished by local application of 1 mm bicuculline. The feedforward IPSP was followed by a delayed EPSP blocked by AP-5 (100 microM), presumably mediated by deep-to-superficial m-EC connections. Our findings demonstrate that superficial m-EC cells are inhibited by the hippocampal output via a feedforward pathway that prevents activity reverberation in the hippocampal-EC-hippocampal loop. We propose that such inhibition could serve as a protective mechanism to prevent epileptic hyperexcitability.

Action Potentials↗

Electrical and chemical long-term depression do not attenuate low-Mg2+-induced epileptiform activity in the entorhinal cortex.

PURPOSE: Low-frequency electrical and magnetic stimulation of cortical brain regions has been shown to reduce cortical excitability and to decrease the susceptibility to seizures in humans and in vivo models of epilepsy. The induction of long-term depression (LTD) or depotentiation of a seizure-related long-term potentiation has been proposed to be part of the underlying mechanism. With the low-Mg(2+)-model of epilepsy, this study investigated the effect of electrical LTD, chemical LTD, and depotentiation on the susceptibility of the entorhinal cortex to epileptiform activity. METHODS: The experiments were performed on isolated entorhinal cortex slices obtained from adult Wistar rats and mice. With extracellular recording techniques, we studied whether LTD induced by (a) three episodes of low-frequency paired-pulse stimulation (3 x 900 paired pulses at 1 Hz), and by (b) bath-applied N-methyl-D-aspartate (NMDA, 20 microM) changes time-to-onset, duration, and frequency of seizure-like events (SLEs) induced by omitting MgSO(4) from the artificial cerebrospinal fluid. Next we investigated the consequences of depotentiation on SLEs themselves by applying low-frequency stimulation after onset of low-Mg(2+)-induced epileptiform activity. RESULTS: LTD, induced either by low-frequency stimulation or by bath-applied NMDA, had no effect on time-to-onset, duration, and frequency of SLEs compared with unconditioned slices. Low-frequency stimulation after onset of SLEs did not suppress but induced SLEs that lasted for the time of stimulation and were associated with a simultaneous increase of the extracellular K(+) concentration. CONCLUSIONS: Our study demonstrates that neither conditioning LTD nor brief low-frequency stimulation decreases the susceptibility of the entorhinal cortex to low-Mg(2+)-induced epileptiform activity. The present study does not support the hypothesis that low-frequency brain stimulation exerts its anticonvulsant effect via the induction of LTD or depotentiation.

Animals↗

A neural circuit analysis of visual recognition memory: role of perirhinal, medial, and lateral entorhinal cortex.

Using a continuous recognition memory procedure for visual object information, we sequentially presented rats with eight novel objects and four repeated objects (chosen from the 8). These were selected from 120 different three-dimensional objects of varying sizes, shapes, textures, and degree of brightness. Repeated objects had lags ranging from 0 to 4 (from 0 to 4 different objects between the first and repeated presentation). An object was presented on one side of a long table divided in half by an opaque Plexiglas guillotine door, and the latency between opening the door and the rat moving the object was measured. The first presentation of an object resulted in reinforcement, but repeated presentations did not result in a reinforcement. After completion of acquisition training (significantly longer latencies for repeated presentation compared with the first presentation of an object), rats received lesions of the perirhinal, medial, or lateral entorhinal cortex or served as sham operated controls. On the basis of postsurgery testing and additional tests, the results indicated that rats with perirhinal cortex lesions had a sustained impairment in performing the task. There were no sustained deficits with medial or lateral entorhinal cortex lesions. The data suggest that recognition memory for visual object information is mediated primarily by the perirhinal cortex but not by the medial or lateral entorhinal cortex.

Animals↗

Medial septal area lesions disrupt theta rhythm and cholinergic staining in medial entorhinal cortex and produce impaired radial arm maze behavior in rats.

This study was designed to determine (1) which brain area paces the theta rhythm in the medial entorhinal cortex (MEC) of rats and (2) the extent to which the behavioral effects of lesions in the medial septal area (MSA), which disrupt the cholinesterase-related pathway to the hippocampal formation, resemble the effects previously reported to result from fimbria-fornix lesions. MSA lesions abolished or decreased theta rhythm in dorsal hippocampus (DHPC) and MEC; acetylcholinesterase (AChE) staining was depleted or diminished in all of the hippocampus and entorhinal cortex. Rats with MSA lesions were impaired on acquisition of a radial arm maze task. Unilateral fimbria lesions left theta rhythm and AChE staining essentially unaltered in ipsilateral DHPC and MEC but depleted AChE in ipsilateral ventral hippocampus (VHPC) and ventral lateral entorhinal cortex (LEC). A lesion of the dorsal fornix at the level of the hippocampal flexure left ipsilateral DHPC theta rhythm and AChE stain unaltered while causing a substantial reduction in theta rhythm and depletion of AChE in ipsilateral MEC. AChE staining was complete in VHPC and LEC. These results suggest tha MSA paces MEC theta rhythm and that the presumed cholinergic projection which mediates this function travels in the dorsal fornix. The fimbria carries a presumed cholinergic projection to ventral LEC. Rats with MSA lesions can learn a radial arm maze task, unlike rats with fimbria-fornix lesions, but they learn significantly slower than normal rats.

Acetylcholinesterase↗

Excitotoxic lesions of the entorhinal cortex leave gustatory within-event learning intact.

The ability of rats with ibotenate lesions of the entorhinal cortex to form memories for events was assessed by using a gustatory within-event learning procedure. Rats first received exposure to 2 events, AX and BY, each composed of a pair of flavors. Following this exposure period, Flavor X alone was paired with the delivery of lithium chloride. Lesioned and control rats showed a greater aversion to A than to B and to AX than to BX. These results challenge theories that suppose that the entorhinal cortex plays a general role in forming representations of patterns of stimulation.

Analysis of Variance↗