Induction of long-term potentiation at perforant path dentate synapses does not affect place learning or memory

In two experiments the authors failed to detect an effect of inducing bilateral, long-lasting synaptic potentiation at perforant path dentate synapses on spatial learning by rats in the Morris place navigation task. Daily sessions of high-frequency stimulation of perforant path axons produced large increases to an asymptotic level in population spike and field excitatory postsynaptic potential recorded in ipsilateral dentate gyrus. Place learning proceeded normally 24 hours after the last of 14 high-frequency stimulation sessions in rats that had previously mastered the procedural aspects of place navigation (Experiment 1) and in rats that were naive (Experiment 2).     

Limbic seizures, but not kindling, reversibly impair place learning in the Morris water maze

We investigated the effects of kindling and kindled seizures in different limbic structures on place and cue learning in the Morris water maze. The triggering of seizures by stimulation of the perforant path, septum, or amygdala prior to daily training impaired place learning, but had little effect on visible platform training or swim speed. Seizures triggered by stimulation of the medial perforant path after daily training also impaired place learning. Conversely, place learning proceeded normally in rats tested 24 h after kindling triggered by stimulation of the perforant path, septum, or amygdala, indicating that kindling per se does not affect place learning. Each group was able to learn the location of a reversed platform when pretraining seizures were discontinued; and perforant path and septal kindled rats, but not amygdaloid kindled rats, were impaired at learning the location of a reversed platform when seizures were triggered before training. The results confirm previous reports that limbic seizures produce amnesia, but they contradict the finding that hippocampal kindling impairs learning on tasks sensitive to hippocampal lesions.     

Neuronal plasticity associated with learning and epileptic seizures: LTP and KIP]

Long-term synaptic potentiation (LTP) and kindling-induced potentiation (KIP) are hypothesized to play an important role in spatial learning and kindling development, respectively, and the possible roles of LTP in spatial learning and KIP in kindling development are reviewed in this paper. Blockage of NMDA receptors, protein synthesis inhibition and knockout of alpha-CaMKII gene markedly impaired both LTP-induction and spatial learning, and destruction of the dentate granule cells with colchicine has been reported to result in severe spatial learning deficits. These findings support the hypothesis that spatial learning may depend on the neuronal input from the entorhinal cortex to dentate granule cells via perforant path and LTP-induction at perforant path-dentate granule cell synapses. However, recent studies have revealed that MPC17742, a selective NMDA receptor antagonist, and 1S, 3S-ACPD, the group II metabotropic glutamate receptor agonist, block LTP-induction at perforant path-dentate granule cell synapses, but that those drugs did not prevent rats from spatial learning. Thus, adaptable changes in the dentate granule cell discharge caused by the neuronal information from the entorhinal cortex are necessary, but LTP at perforant path-dentate granule cell synapses is not necessarily requisite for spatial learning. It has been also hypothesized that kindling development might be based on the long-lasting synaptic potentiation (the KIP/kindling hypothesis). Destruction of the dentate granule cells with colchicine retarded kindling development of amygdala or entorhinal cortex has been reported, and repeated induction of LTP at perforant path-dentate granule cell synapses, furthermore, caused anomalous mossy fiber sprouting and facilitated the subsequent kindling development. These results are in accordance with the KIP/kindling hypothesis. However, even when LTP was induced once a day for 20 days, the repeated induction of LTP failed to induce epileptic discharge. We demonstrated that KIP observed in an interictal period faded away gradually during kindling stimulation before epileptic seizures began. Furthermore, rapid kindling at an interstimulus interval of 5 min blocked completely the development of KIP, whereas the afterdischarge prolonged gradually and generalized convulsions were often observed during the late stage of rapid kindling. Thus, LTP and KIP are not indispensable for kindling development, even if LTP facilitate the subsequent kindling development. It should be noted that instead of KIP, the abnormal plasticity essential for kindling development must appear during an transition period from interictal to ictal periods.     

Enhanced long-term potentiation induced in rat dentate gyrus by coactivation of septal and entorhinal inputs: Temporal constraints

High-frequency activation of the entorhinal cortical (perforant path) inputs to the rat dentate gyrus can produce a long-term potentiation (LTP) of perforant path-dentate evoked responses. In this paper we examined the enhanced LTP effects produced by coactivation of septal and entorhinal inputs to the dentate gyrus. Trains of electrical stimulation applied to the two inputs were found to increase the magnitude of LTP to a level above that produced by trains applied to the perforant path alone. The largest LTP increments were observed when the septal trains were applied less than 100 ms prior to the perforant path trains. If the septal trains followed the perforant path trains there was no additional increment in LTP magnitude, regardless of the intertrain interval. The relationship of this cooperativity effect to mechanisms of associative learning is discussed.     

Medial and lateral perforant path evoked potentials are selectively modulated by pairing with glutamatergic activation of locus coeruleus in the dentate gyrus of the anesthetized rat

Norepinephrine (NE) in vitro produces long-lasting potentiation of medial perforant path input and depression of lateral perforant path input to dentate gyrus in the rat. Similar, but highly transient, effects have been reported in vivo using paragigantocellular stimulation to release NE. The present study uses alternate stimulation of the medial perforant path and lateral olfactory tract (eliciting a lateral perforant path-evoked potential) to examine the effects of glutamatergic activation of locus coeruleus (LC) on the two pathways for up to 3 h post-LC activation. In the first experiment, the expected potentiation of the medial perforant path population spike in dentate gyrus was observed, but without accompanying depression of the lateral perforant path-mediated evoked potential (lateral olfactory tract stimulation, 60 s ISI). In a second experiment, with more frequent pairing of input with NE release (10 s ISI), significant potentiation of lateral perforant path-mediated input to dentate gyrus occurred, but potentiation of medial perforant path input was not seen. A third experiment with a 30 s ISI again produced potentiation of lateral perforant path-mediated input without potentiation of the medial perforant path population spike. The size of effects with the 30 s ISI was intermediate between that seen with 10 s and 60 s ISI. Potentiation of lateral perforant path over medial perforant path input has previously been reported with acute nicotinic activation of the LC. This outcome also resembles heterosynaptic modulation previously reported with tetanic potentiation. The data argue for a competitive relationship between medial and lateral perforant path inputs to dentate gyrus and suggest pairing with increased NE produces a bias favoring one or the other pathway depending on parameters such as strength and frequency. NE potentiating effects on lateral perforant path input here may also have occurred in entorhinal cortex (EC) given the system-wide NE release with LC activation.     

Effects of synaptic antagonists on perforant path paired-pulse plasticity: differentiation of pre- and postsynaptic antagonism

The effects of different synaptic antagonists on paired-pulse plasticity of medial perforant path responses were studied in rat hippocampal slices. Baclofen reduces the response to activation of the perforant path, but does not have the same net effect on the first and second responses to paired stimulation: baclofen lessens the percent paired-pulse depression of medial perforant path responses. Furthermore, at doses that reduced the control medial perforant path response by half, paired-pulse plasticity changed from paired-pulse depression to paired-pulse potentiation. A similar effect on medial perforant path paired-pulse plasticity is produced by decreasing extracellular calcium concentration. Kynurenic acid reduces the first and second responses to paired stimulation proportionately the same, and, therefore, has no effect on the percent paired-pulse depression. These results suggest that baclofen acts presynaptically to reduce the synaptic response, whereas kynurenate acts postsynaptically. Adenosine was also found to be a potent antagonist of medial perforant path responses, with effects on paired-pulse plasticity similar to baclofen: a new synaptic antagonist, N-p-chlorobenzoyl-piperazine-2,3-dicarboxylate, was found to have effects like kynurenate, suggesting that it is also a postsynaptic receptor blocker.     

Behavioural, electrophysiological and histopathological changes following sustained stimulation of the perforant pathway input to the hippocampus: Effect of the NMDA receptor antagonist, CGP 39551

Sustained stimulation of the perforant path has been shown to damage the CA1 area and impair spatial memory in rats. The pattern of cell death is similar in human epileptics, who also exhibit memory deficits. In this study we demonstrate that the learning/memory impairment in water maze test and the development of interictal spikes that also followed stimulation-induced damage were antagonized by CGP 39551. Pretreatment with this NMDA receptor antagonist also slightly diminished somatostatin cell loss in the hilus but not CA1 pyramidal cell damage. These results indicate that the impairment of spatial learning/memory seems to be dependent not only on the degree of cell degeneration in the CA1 subfield of the hippocampus but also on the frequency of interictal spikes, at least in this model of epilepsy.     

Stimulation-induced reset of hippocampal theta in the freely performing rat

Previous research has suggested that visual and auditory stimuli in a working memory task have the ability to reset hippocampal theta, perhaps allowing an organism to encode the incoming information optimally. The present study examined two possible neural pathways involved in theta resetting. Rats were trained on a visual discrimination task in an operant chamber. At the beginning of a trial, a light appeared over a centrally located lever that the rat was required to press to receive a water reward. There was a 30-s intertrial interval before the next light stimulus appeared. After learning the task, all rats received surgical implantation of stimulating electrodes in both the fornix and the perforant path and recording electrodes, bilaterally in the hippocampus. After surgery, theta was recorded before and after the light stimulus to determine whether resetting to the visual stimulus occurred. During the intertrial interval, rats received single-pulse electrical stimulation of either the fornix or perforant path. Theta was recorded both before and after the electrical stimulation to determine whether resetting occurred. In this experiment, hippocampal theta was reset after all three stimulus conditions (light, perforant path, and fornix stimulation), with the greatest degree of reset occurring after the fornix stimulation. The results suggest that activation of the perforant path and fornix may underlie theta reset and provide a mechanism by which the hippocampus may enhance cognitive processing.     

Lateral olfactory tract input to dentate gyrus is depressed by prior noradrenergic activation using nucleus paragigantocellularis stimulation

Nucleus paragigantocellularis stimulation potentiates the medial perforant path population spike in the dentate gyrus via β-receptor activation. In this study, the same paragigantocellularis stimulation preceding lateral olfactory tract pulses depressed the lateral perforant path mediated synaptic potential in dentate gyrus. Depression of the lateral olfactory tract input was blocked by a β-antagonist. These in vivo results confirm in vitro reports that norepinephrine induces potentiation of medial perforant path input and depression of lateral perforant path input to dentate gyrus.     

An analysis of the increase in granule cell excitability accompanying habituation in the dentate gyrus of the anesthetized rat

Repeated low-frequency stimulation of the perforant path results in a decrement in the population EPSP and population spike recorded in the hilus of the dentate gyrus. The EPSP decrement is accompanied, however, by an increase in the population spike height/population EPSP slope relation, suggesting that an increase in granule cell excitability also occurs. The present experiments explored the mechanisms of this apparent increase in excitability using standard field potential recording techniques to assess perforant path input/output curves in rats anesthetized with sodium pentobarbital. Low-frequency homosynaptic stimulation (512 pulses, 1 Hz) of the perforant path resulted in a decreased spike threshold and overall shift to the left of the function relating population spike height to EPSP slope. These changes were consistently produced, even when granule cell discharge was inhibited by conditioning stimulation of the contralateral hilus. On the other hand, low-frequency heterosynaptic (lateral perforant path) or antidromic (mossy fiber) driving of the granule cells only slightly increased the medial path spike/EPSP relation, and did not alter the spike threshold. The excitability shift accompanying habituation was qualitatively different from that associated with long-term potentiation, but these shifts did not summate. The interpretation which best explains these various results is that granule cell excitability is increased during low-frequency perforant path stimulation by a process of disinhibition, caused by habituation of perforant path excitatory synaptic drive onto feed-forward inhibitory interneurons.     

Functional deficits after sustained stimulation of the perforant path

Several reports have implicated the overactivity of hippocampal glutaminergic systems in neurodegenerative conditions including Senile dementia of the Alzheimer's type (SDAT). The neurobiological effects of hippocampal glutaminergic hyperactivity were studied by perforant pathway stimulation. Forty-five minutes of sustained perforant pathway stimulation produced a 50% or greater increase in motor activity 1, 2, and 3 weeks after stimulation. Robust retention deficits in a 48-h step-through passive avoidance task were evident 2 weeks post-stimulation. Furthermore, animals receiving stimulation were impaired in the acquisition of a spatial task in the Morris water maze. Stimulated animals exhibited little reduction in their escape latencies over the testing period. The learning and memory deficits were associated with a loss of CA1 and CA3 pyramidal cells and pretreatment with the N-methyl-D-aspartate antagonist MK-801 reduced this cell loss, particularly in the CA1 region of the hippocampus. These results suggest that sustained stimulation of the perforant pathway may be useful in studying neurological deficits associated with glutaminergic hyperfunction.     

Incorporation of [H]fucose in rat hippocampal structures after conditioning by perforanth path stimulation and after LTP-producing tetanization

The contribution of glycoprotein synthesis to functional synaptic changes and to the formation of memory traces was investigated by autoradiographic determination of the incorporation of [³H]fucose into the hippocampal structures of rats. In the first experiment, the fucose incorporation was measured after induction of post-tetanic long-term potentiation (LTP) in granular cell synapses by repeated tetanization (200 cps) of the perforant path, and after stimulation of this hippocampal input by the same number of impulses with very low frequency (0.2 cps) not producing LTP. In the second experiment, the incorporation of fucose was determined after an active avoidance training using the stimulation of the perforant path by impulse trains of 15 cps as conditioning stimuli, and after a session of corresponding unpaired stimulations of the perforant path. Unstimulated animals were used in both experiments to measure the basal glycosylation. LTP-producing tetanization resulted only in a slight increase of incorporation into the ipsilateral hippocampal structures without significant differences to similar changes after the corresponding control stimulation with single impulses. After a session of unpaired stimulation of the perforant path with impulse trains of 15 cps only slight and inconsistent changes of incorporation occurred in the hippocampus too. However, after conditioning by the corresponding perforant path stimulation as conditioned stimulus, considerable increases of incorporation were observed in all structures of the ipsilateral hippocampus, when compared to the unpaired control stimulation. An enhanced labeling occurred also in some structures of the contralateral hippocampus mainly receiving commissural inputs. The results suggest again, that the activation of one single hippocampal afferent, even if producing LTP, would not be sufficient to induce an increased glycosylation of neuronal proteins. The increase of glycoprotein formation seems to require the convergence of several inputs, which can be assumed to occur during learning. Therefore, LTP of a single synaptic population seems not to represent the complete long-lasting memory trace, but only one of its components, or a preceding transient storage mechanism.     

Physiological and Morphological Heterogeneity of Dentate Gyrus—Hilus Interneurons in the Gerbil Hippocampus In Vivo

A variety of morphological types of dentate gyrus/hilus interneurons have been described, but little is known about their corresponding physiological characteristics. To address this issue, intracellular responses to current injection and perforant path stimulation were obtained from putative dentate interneurons in anaesthetized adult gerbils. Our sample of interneurons showed heterogeneity in their intrinsic physiological characteristics and spike thresholds to perforant path stimulation, suggesting the existence of distinct physiologically-defined classes. 'Fast-spiking'interneurons had a low threshold to perforant path stimulation, whereas 'slow-spiking'interneurons responded with predominantly inhibitory potentials. In several cases, cells were intracellularly labelled with biocytin for visualization. Interneurons with different physiological traits had distinct morphological features. These results confirm that, as in hippocampus proper, morphologically identifiable interneurons in the dentate hilus show electrophysiological features that are likely to reflect functionally specific roles in informational processing.     

Characterization of perforant path lesions in rodent models of memory and attention

Early stage Alzheimer's disease (AD) pathology is associated with neurodegeneration of systems within the temporal cortex, e.g. the entorhinal cortex, perforant pathway and hippocampus. The perforant pathway provides the major neuronal input to the hippocampus from the entorhinal cortex and thus relays multimodal sensory information derived from cortical zones into the hippocampus. The earliest symptoms of AD include cognitive impairments, e.g. deficits in short-term memory and attention. Consequently, we have investigated the effect of bilateral knife cut lesions to the perforant path on cognition in rats using models measuring primarily short-term memory (operant delayed match to position task), attention (serial five-choice reaction time task) and spatial learning (Morris water maze). Rats receiving bilateral perforant path lesions showed normal neurological function and a mild hyperactivity. The lesion produced little effect on attention assessed using the five-choice task. In contrast, animals with equivalent lesions showed a robust delay-dependent deficit in the delayed match to position task. Spatial learning in the water maze task was also severely impaired. The delay-dependent deficit in the match to position task was not reversed by tacrine (3 mg/kg) pretreatment. The present data support a selective impairment of cognitive function following perforant path lesions that was confined to mnemonic rather than attentional processing. These findings complement primate and human studies identifying a critical role of the perforant pathway and associated temporal lobe structures in declarative memory. Degeneration of the perforant pathway is likely to contribute to the mnemonic deficits characteristic of early AD. The failure of tacrine to ameliorate these deficits may be relevant to an emerging clinical literature suggesting that cholinomimetic therapies improve attentional rather than mnemonic function in AD.     

Comparative characteristics of the direct influences of the perforant pathway on neurons in hippocampal fields CA1 and CA3 in vitro]

The effect of the perforant path stimulation on the CA1 and CA3 neurons was investigated in incubated slices of the guinea pig hippocampus. Spike generation was observed in both fields during stimulation of the perforant path. The majority of the CA1 neurons followed rhythmic stimulation up to 30-80/c. The CA3 neurons responded only to low-frequency stimulation (up to 5/c). The posttetanic potentiation of responses to the perforant path stimulation was observed in both hippocampal fields.     

Long-term effects of transcranial magnetic stimulation on hippocampal reactivity to afferent stimulation.

Transcranial magnetic stimulation (TMS) has become a promising treatment of affective disorders in humans, yet the neuronal basis of its long-lasting effects in the brain is still unknown. We studied acute and lasting effects of TMS on reactivity of the rat hippocampus to stimulation of the perforant path. Application of TMS to the brain of the anesthetized rat caused a dose-dependent transient increase in population spike (PS) response of the dentate gyrus to perforant path stimulation. In addition, TMS caused a marked decrease in inhibition and an increase in paired-pulse potentiation of reactivity to stimulation of the perforant path. Also, TMS suppressed the ability of fenfluramine (FFA), a serotonin releaser, to potentiate PS response to perforant path stimulation. Chronic TMS did not affect single population spikes but caused an increase in paired-pulse potentiation, which was still evident 3 weeks after the last of seven daily TMS treatments. After chronic TMS, FFA was ineffective in enhancing reactivity to perforant path stimulation, probably because it lost the ability to release serotonin. In addition, the beta adrenergic receptor agonist isoproterenol, which caused an increase in PS in the control rats, failed to do so in the TMS-treated rats. These results indicate that TMS produces a long-term reduction in efficacy of central modulatory systems.     

Perforant path stimulation in rats produces seizures, loss of hippocampal neurons, and a deficit in spatial mapping which are reduced by prior MK-801

Severe temporal lobe epilepsy in humans is often associated with loss of neurons in the hippocampus and memory deficits. In Experiment 1, 60 min of continuous electrical stimulation of the perforant path sufficient to produce seizures resembling status epilepticus and loss of hilar and pyramidal cells in the hippocampus, produced a deficit in spatial mapping in the Morris water tank. In particular, the previously stimulated rats took longer and swam farther to find a hidden, but not a visually cued, platform, and, in contrast to the unstimulated control rats, were not disrupted by movement of the platform to a new location. In Experiment 2, a single injection of the non-competitive NMDA receptor antagonist, MK-801 (1.0 mg/kg), just prior to the perforant path stimulation reduced the seizures, hippocampal neuronal loss, and deficit in spatial mapping. These data suggest that temporal lobe seizures can induce deficits in spatial memory by selectively destroying neurons within the hippocampus, and that the mechanism by which this occurs involves the activation of NMDA receptors, and, perhaps, consequent excitotoxicity.     

Evoked potentials and long-term potentiation in the mouse dentate gyrus after stimulation of the entorhinal cortex

When the entorhinal cortex is electrically stimulated, an evoked potential is produced in the ipsilateral dentate gyrus of the mouse which is similar to this response in the rat, rabbit, and other preparations in which it has been recorded. The evoked potential consists of two components: Component I represents an extracellular monosynaptic population EPSP, and Component II represents a population spike due to the synchronous activation of the granule cells. After tetanic stimulation, responses to single-pulse stimuli increased and remained at that level for at least 30 min. This demonstrated long-term potentiation of the evoked potential, phenomenologically similar to that described in other species. A medial and lateral perforant path could be differentiated. Stimulation of the medial perforant path resulted in evoked potentials that had a short peak latency, half-width, and rise time. Conversely, stimulation of the lateral perforant path resulted in evoked potentials that had a long peak latency, half-width, and rise time. The recording electrode depth series were also different after selective activatio of the two paths. Stimulation of the medial perforant path resulted in recording electrode depth series that had their maximum negativities close to the cell body layer, whereas stimulation of the lateral perforant path resulted in depth series that had their maximum negativities farther out along the granule cell dendrites. These results are consistent with the different sites of termination of the two paths. Long-term potentiation was demonstrated in both pathways after administration of tetanic stimuli of various parameters.     

Long-term potentiation recruits a trisynaptic excitatory associative network within the mouse dentate gyrus

Granule cells of the hippocampal dentate gyrus receive two powerful excitatory inputs: the perforant path, originating from the entorhinal cortex, and the associational pathway, originating from mossy cells, the principal neurons of the dentate gyrus hilus. We examined the electrophysiological properties of the less well-studied associational pathway and its interaction with the perforant path in the intact mouse hippocampus and then tested homosynaptic, trans-synaptic and associative long-term potentiation of these pathways. The associational pathway was either monosynaptically activated by stimulation within the inner molecular layer or trisynaptically activated after stimulation of the perforant path. Laminar profiles of extracellularly recorded associational pathway field potentials demonstrated a bell-shaped curve with a peak in the inner molecular layer. Tetanization of the perforant path induced not only homosynaptic potentiation of the perforant path (162.4 +/- 6.7% at 0.5-1.5 h after tetanus) but also heterosynaptic potentiation of the associational pathway (115.7 +/- 4.9%). Direct tetanization of the associational pathway within the inner molecular layer was ineffective in either the septo-temporal (97.2 +/- 4.5%) or temporal-septal (104.4 +/- 4.6%) direction. In contrast, conjoint tetanization of the associational pathway with the perforant path potentiated the associational pathway responses in both the septo-temporal (123.4 +/- 5.8%) and the temporal-septal (124.8 +/- 7.3%) directions. Paired-pulse facilitation was attenuated by long-term potentiation in the perforant path and the associational pathway, suggesting pre-synaptic involvement. These results demonstrate that long-term potentiation of the associational pathway and the perforant path is a product of the network properties of the dentate gyrus rather than of each monosynaptic input alone. The architecture of this neural network may be designed for flexible dynamic associations of the afferent perforant path inputs to configure encoded information within hippocampal neuronal ensembles.     

Permanently altered hippocampal structure, excitability, and inhibition after experimental status epilepticus in the rat: the "dormant basket cell" hypothesis and its possible relevance to temporal lobe epilepsy.

The relationship between an episode of status epilepticus, the resulting hippocampal pathology, and the subsequent development of pathophysiological changes possibly relevant to human epilepsy was explored using the experimental epilepsy model of perforant path stimulation in the rat. Granule cell hyperexcitability and decreased feedforward and feedback inhibition were evident immediately after 24 hours of intermittent perforant path stimulation and persisted relatively unchanged for more than 1 year. All of the pathophysiological changes induced by perforant path stimulation were replicated in normal animals by a subconvulsive dose of bicuculline, suggesting that the permanent "epileptiform" abnormalities produced by sustained perforant path stimulation may be due to decreased GABA-mediated inhibition. Granule cell pathophysiology was seen only in animals that exhibited a loss of adjacent dentate hilar mossy cells and hilar somatostatin/neuropeptide Y-immunoreactive neurons. GABA-immunoreactive dentate basket cells survived despite the extensive loss of adjacent hilar neurons. However, parvalbumin immunoreactivity, present normally in a subpopulation of GABA-immunoreactive dentate basket cells, was absent on the stimulated side. Whether this represents decreased parvalbumin synthesis in surviving basket cells or a loss of a specific subset of inhibitory cells is unclear. Hyperexcitability and decreased paired-pulse inhibition in response to ipsilateral perforant path stimulation were also present in the CA1 pyramidal cell layer on the previously stimulated side, despite minimal damage to CA1 pyramidal cells or interneurons. The possibility that CA1 inhibitory neurons were hypofunctional or "dormant" due to a loss of excitatory input to inhibitory cells from damaged CA3 pyramidal cells was tested by stimulating the contralateral perforant path in order to activate the same CA1 basket cells via different inputs. Contralateral stimulation evoked CA1 pyramidal cell paired-pulse inhibition immediately in the previously stimulated hippocampus. Thus, we propose the "dormant basket cell" hypothesis, which implies that despite malfunction, inhibitory systems remain intact in "epileptic" tissue and are capable of functioning if appropriately activated.