Extrahippocampal Structures Research Articles

Parallel activation of field CA2 and dentate gyrus by synaptically elicited perforant path volleys.

Previous studies showed that dorsal psalterium (PSD) volleys to the entorhinal cortex (ENT) activated in layer II perforant path neurons projecting to the dentate gyrus. The discharge of layer II neurons was followed by the sequential activation of the dentate gyrus (DG), field CA3, field CA1. The aim of the present study was to ascertain whether in this experimental model field, CA2, a largely ignored sector, is activated either directly by perforant path volleys and/or indirectly by recurrent hippocampal projections. Field potentials evoked by single-shock PSD stimulation were recorded in anesthetized guinea pigs from ENT, DG, fields CA2, CA1, and CA3. Current source-density (CSD) analysis was used to localize the input/s to field CA2. The results showed the presence in field CA2 of an early population spike superimposed on a slow wave (early response) and of a late and smaller population spike, superimposed on a slow wave (late response). CSD analysis during the early CA2 response showed a current sink in stratum lacunosum-moleculare, followed by a sink moving from stratum radiatum to stratum pyramidale, suggesting that this response represented the activation and discharge of CA2 pyramidal neurons, mediated by perforant path fibers to this field. CSD analysis during the late response showed a current sink in middle stratum radiatum of CA2 followed by a sink moving from inner stratum radiatum to stratum pyramidale, suggesting that this response was mediated by Schaffer collaterals from field CA3. No early population spike was evoked in CA3. However, an early current sink of small magnitude was evoked in stratum lacunosum-moleculare of CA3, suggesting the presence of synaptic currents mediated by perforant path fibers to this field. The results provide novel information about the perforant path system, by showing that dorsal psalterium volleys to the entorhinal cortex activate perforant path neurons that evoke the parallel discharge of granule cells and CA2 pyramidal neurons and depolarization, but no discharge of CA3 pyramidal neurons. Consequently, field CA2 may mediate the direct transfer of ENT signals to hippocampal and extrahippocampal structures in parallel with the DG-CA3-CA1 system and may provide a security factor in situations in which the latter is disrupted.

Vigabatrin protects against hippocampal damage but is not antiepileptogenic in the lithium-pilocarpine model of temporal lobe epilepsy

In temporal lobe epilepsy (TLE), the nature of the structures involved in the development of the epileptogenic circuit is still not clearly identified. In the lithium–pilocarpine model, neuronal damage occurs both in the structures belonging to the circuit of initiation and maintenance of the seizures (forebrain limbic system) as well as in the propagation areas (cortex and thalamus) and in the circuit of remote control of seizures (substantia nigra pars reticulata). In order to determine whether protection of some brain areas could prevent the epileptogenesis induced by status epilepticus (SE) and to identify the cerebral structures involved in the genesis of TLE, we studied the effects of the chronic exposure to Vigabatrin (gamma-vinyl-GABA, GVG) on neuronal damage and epileptogenesis induced by lithium-pilocarpine SE. The animals were subjected to SE and GVG treatment (250 mg/kg) was initiated at 10 min after pilocarpine injection and maintained daily for 45 days. These pilo–GVG rats were compared with rats subjected to SE followed by a daily saline treatment (pilo–saline) and to control rats not subjected to SE (saline–saline). GVG treatment induced a marked, almost total neuroprotection in CA3, an efficient protection in CA1 and a moderate one in the hilus of the dentate gyrus while damage in the entorhinal cortex was slightly worsened by the treatment. All pilo–GVG and pilo–saline rats became epileptic after the same latency. Glutamic acid decarboxylase (GAD67) immunoreactivity was restored in pilo–GVG rats compared with pilo–saline rats in all areas of the hippocampus, while it was increased over control levels in the optical layer of the superior colliculus and the substantia nigra pars reticulata. Thus, the present data indicate that neuroprotection of principal cells in the Ammon's horn of the hippocampus is not sufficient to prevent epileptogenesis, suggesting that the hilus and extra-hippocampal structures, that were not protected in this study, may play a role in the genesis of spontaneous recurrent seizures in this model. Furthermore, the study performed in non-epileptic rats indicates that chronic treatment with a GABAmimetic drug upregulates the expression of the protein GAD67 in specific areas of the brain, independently from the seizures.

Blockade of Excitability of Hippocampal Neurons after Coagulation of the Dorsal Amygdalofugal Pathway

In chronic experiments on rabbits, we demonstrated that electrolytic destruction of the stria terminalis results in the intense depression of the integral electrical activity of the hippocampus. Characteristics of the responses induced by electrical and chemostimulation of the hippocampus under the above conditions allowed us to conclude that a significant drop in the excitability of hippocampal neurons themselves, but not changes in the structure and intensity of afferent impulsation from the extrahippocampal structures, underlies such a depression. Morphological changes in neurons and satellite glial cells, such as lysis of the Nissl substance, hyperchromatosis, and wrinkling of the cells after destruction of the stria terminalis, were proof of the correctness of our conclusion. It is postulated that one of the main factors underlying the generalized effects observed in our experiments is probably the disturbance of functional characteristics of the hypothalamo-hypophyseal neurosecretory system induced by destruction of the amygdalofugal connections; this disturbance results in significant pathological shifts of metabolism in the hippocampus.

Acute Administration of Estrogen and Progesterone Impairs the Acquisition of the Spatial Morris Water Maze in Ovariectomized Rats

Although several markers of synaptic efficacy are enhanced during proestrus, spatial water maze performance is impaired. Because levels of both estrogen and progesterone are elevated in proestrus, the nature of their individual and combined effects on spatial learning was examined. Long-Evans hooded rats were ovariectomized postpubertally and pretrained on a water maze with a visible platform (nonspatial). Following pretraining, rats were administered estrogen (5 μg sc) or oil 48 and 24 h prior to testing and progesterone (500 μg sc) or oil 4 h prior to testing. Rats were tested on a water maze in a different room with a submerged platform (spatial) for 16 trials with random start location in a single testing day. Latency and path length to the target platform were significantly greater in estrogen plus progesterone-treated animals than in controls. Neither estrogen nor progesterone alone significantly impaired performance relative to controls on either measure. Swim speed was not significantly affected by any of the hormone treatments. Performance on a nonspatial cue task was not significantly altered by ovarian steroids. Thus, the combination of estrogen and progesterone produces deficits in the acquisition of the Morris water maze that may be specific to spatial reference memory. These deficits could be due to hormonal influences on extrahippocampal structures or to detrimental effects on behavior resulting from the increased synaptic activity intrinsic to the hippocampus proper.

Epileptogenesis after status epilepticus reflects age- and model-dependent plasticity

Although epilepsy often begins in childhood, factors that contribute to the development of epilepsy as a consequence of status epilepticus (SE) during early development are poorly understood. We investigated animal models in which seizure-induced epileptogenicity could be studied. Rats undergoing self-sustaining SE induced by perforant path stimulation (PPS) at the ages of postnatal day 21 (P21) and P35 were compared with those subjected to SE by lithium and pilocarpine (LiPC). Although only one animal subjected to PPS at P21 developed chronic spontaneous seizures by several months of observation, all the animals subjected to PPS at P35 became epileptic. In the LiPC model, however, most of the rat pups subjected to SE at P21 became epileptic. Animals with spontaneous seizures showed increased inhibition in the dentate gyrus, a characteristic of the epileptic brain, with evidence of mossy fiber synaptic reorganization. Examination of circuit recruitment by c-Jun immunohistochemistry showed activation restricted to the hippocampus in P21 animals subjected to PPS, although extensive activation of hippocampal and extrahippocampal structures was seen in pups subjected to PPS-induced self-sustaining SE at P35 or LiPC SE at P21. These results demonstrate that the appearance of epilepsy as a consequence of SE is influenced by the type of insult as well as by age-dependent circuit recruitment.

El diagnóstico de la esclerosis temporal mesial mediante imagen de resonancia magnética

Mesial temporal sclerosis (MTS) is a progressive drug-resistant epileptic syndrome which requires rapid, effective diagnosis and treatment. Histologically there is atrophy and gliosis of the hippocampus. To establish magnetic resonance (MR) imaging guidelines for correct diagnosis. We made a prospective study of 78 patients with drug-resistant temporal lobe epilepsy (44 women and 34 men; age 6-66 years, mean 31 years). Using a magnet of 1.5 Teslas paracoronal sections were made of the hippocampus with T1 volumetric with inversion-recovery, FLAIR (fluid-attenuated inversion-recovery) and T2 relaxometry. A control group of 30 healthy volunteers was established. The reduction in volume and hippocampal T2 hyperintensity were considered to be MTS diagnosed on MR. No hippocampal differences were observed among the healthy volunteers. The confidence intervals (mean +/- 1.96 SD) were: right volume: 4.169-5.911 mm3; left volume: 4.097-5.940 mm3; time of T2 relaxation: 98-113 ms. MTS was observed in 42 patients (54%): 24 left, 14 right and four asymmetrical bilateral. The results of the diagnostic validity (sensitivity/specificity) were: T1 volumetric 91/92%, FLAIR 93.5/98% and T2 relaxometry 91/92%. There was atrophy of other extrahippocampal structures in five cases of MTS; 10 patients with MTS (23.5%) had another extrahippocampal lesion associated (dual pathology), particularly migration disorders; 21 patients (27%) had lesions without MTS (tumors, alterations of migration, nonspecific gliosis) and in 15 cases (19%) there were no abnormal findings. A total of 27 patients were operated on: 22 with MTS (21 had diagnostic MR, one case had no abnormal findings), four cases had tumors and one had cortical dysplasia. The combination of quantitative techniques (T1 volumetric with inversion-recovery and T2 relaxometry) and FLAIR optimize MTS diagnosis using MR.

Electrophysiologic analysis of a chronic seizure model after unilateral hippocampal KA injection.

Unilateral intrahippocampal injections of kainic acid (KA) in rats produce spontaneous recurrent limbic seizures and morphologic changes in hippocampus that resemble hippocampal sclerosis in patients with medically refractory mesial temporal lobe epilepsy (MTLE), that form of temporal lobe epilepsy (TLE) associated with hippocampal sclerosis. Interictal in vivo electrophysiologic studies have revealed high-frequency (250-500 Hz) oscillations, termed fast ripples (FRs). These oscillations may uniquely occur in or adjacent to the site of hippocampal KA injection, in areas that generate spontaneous seizures. Similar field potentials also have been demonstrated in the epileptogenic region of patients with TLE. We have now characterized ictal electrographic patterns in this rat model for comparison with those in human TLE and begun to evaluate the role of FRs in the transition to ictus in the KA-treated rat. Rats received unilateral intrahippocampal injections of KA and, after the development of spontaneous seizures, were implanted with multiple fixed and moveable microelectrodes for single unit, field potential, and EEG recording. They were then monitored by using video-EEG telemetry for several weeks to capture and evaluate electrographic and behavioral seizure types. Results were correlated with Timm's stain demonstration of mossy fiber sprouting. Low-voltage fast (LVF) and hypersynchronous electrographic ictal-onset patterns were seen in the KA-treated rat that resembled similar ictal-onset patterns in patients with TLE. Hypersynchronous, but not LVF, ictal discharges were associated with recurrent FRs. As in the human, hypersynchronous ictal onsets originated predominantly in hippocampus, whereas LVF ictal onsets more often involved extrahippocampal structures. LVF ictal onsets occurred during wakefulness or paradoxical sleep and were usually associated with motor behavior, whereas hypersynchronous ictal onsets occurred during slow-wave sleep or periods of immobility and were not associated with motor behavior unless there was transition to another ictal electrographic pattern. Mossy fiber sprouting did not correlate with the frequency of ictal EEG discharges exhibited by each rat but was greater in those rats that demonstrated frequent behavioral seizures. The electrographic features of spontaneous seizures in the KA-treated rat resemble those of patients with medically refractory TLE with respect to EEG pattern and localization. Our data suggest that hypersynchronous ictal onsets represent epileptogenic disturbances in hippocampal circuits, whereas LVF ictal onsets may involve extrahippocampal areas having more direct connections to the motor system. Hypersynchronous seizures may involve the same neuronal mechanisms that generate interictal FRs.

Effects of Ibotenate Hippocampal and Extrahippocampal Destruction on Delayed-Match and -Nonmatch-to-Sample Behavior in Rats

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

Dissociation of mossy fiber sprouting and electrically-induced seizure sensitivity: rapid kindling versus adaptation.

It has been shown that massed stimulation (MS) of the amygdala or hippocampus does not result in seizure progression but in the 'phenomenon of adaptation', whereas alternate day rapid kindling (ADRK) produces reliable kindling (Lothman, E.W., Williamson, J.M., 1994. Brain Res. 649, 71-84). The goal of the present experiment was to determine if the two different effects are due to differences in mossy fiber sprouting and/or different seizure and postictal spike propagation patterns. Nine rats underwent MS (66-70 stimulations separated by 5-min interstimulus interval), six were exposed to ADRK (12 stimulations/day, every 30 min, with 4 stimulus days, each separated by 1 stimulus-free day), five rats served as control. All rats had electrodes implanted bilaterally in dorsal and ventral hippocampi (VH) and 14 of them had additional electrodes in the piriform cortex. Animals were stimulated in the left VH at afterdischarge threshold. There was no potentiation in seizure response 4-7 weeks after MS. In contrast, ADRK produced not only kindling but also ongoing epileptogenesis resulting 4-7 weeks later in spontaneous seizures and development of a prolonged convulsive state in response to the initially subconvulsive stimulus. Epileptiform activity during MS was mostly restricted to VH, whereas during ADRK it spread widely among studied structures including piriform cortex. Afterdischarges during MS were elicited frequently but seizures did not progress beyond stage 2-3. During ADRK, afterdischarges were evoked less frequently but seizures reached stage 4-7 by the end of the 3rd and 4th stimulus days. The fully kindled state was not reached at this time, but epileptogenic changes continued to progress. Seven weeks after the initial stimulation, both groups demonstrated mossy fiber sprouting of similar intensity in VH. We suggest, (1) frequent but predominantly local hippocampal afterdischarges induce mossy fiber sprouting, but this is not sufficient to produce significant enhancement in seizure susceptibility, and (2) the involvement of extra-hippocampal structures, possibly piriform cortex, and formation of an aberrant hippocampal-para-hippocampal circuit is required to result in a condition of progressive epileptogenesis.

Head direction cells and episodic spatial information in rats without a hippocampus.

To successfully navigate through the environment animals rely on information concerning their directional heading and location. Many cells within the postsubiculum and anterior thalamus discharge as a function of the animal's head direction (HD), while many cells in the hippocampus discharge in relation to the animal's location. We placed lesions in the hippocampus and recorded from HD cells in the postsubiculum and anterior thalamus. Lesions of the hippocampus did not disrupt the HD cell signal in either brain area, indicating that the HD cell signal must be generated by structures external to the hippocampus. In addition, each cell's preferred firing direction remained stable across days when the lesioned animal was placed into a novel environment. This stability appeared to weaken after several weeks of nonexposure to the new enclosure for two out of five animals, and subsequently recorded cells from these two animals established a new angular relationship between the familiar and novel environments. Our results suggest that extra-hippocampal structures are capable of creating and maintaining a novel representation of the animal's environmental context. This representation shares features in common with mnemonic processes involving episodic memory that until now were assumed to require an intact hippocampus.

Physiological evidence for hippocampal disinhibition resulting from activation of the median raphe

The mechanism underlying median raphe (MR)-induced facilitation of hippocampal synaptic transmission was investigated by recording stimulus-evoked field potentials and unitary responses in urethanetized rats. Stimulation of the MR 40 ms prior to perforant path (PP) activation significantly increased the magnitude of PP-evoked granule cell population spikes (median increase = 78%) without affecting population EPSP slope. Injection of homocysteic acid into the vicinity of the MR also facilitated PP-evoked granule cell population spikes, in a dose-dependent manner. Nineteen dentate hilar units were characterized as putative interneurons on the basis of their waveform characteristics and their response to PP stimulation. Electrical activation of the MR inhibited spontaneous or PP-evoked activity in the majority (75%) of these cells: the remaining cells were unaffected. MR stimulation also inhibited spontaneous activity in a large proportion (60%) of putative interneurons in CA1. The present results provide evidence that neurons within the raphe modulate hippocampal throughput by altering discharge of non-principal cells. These data, thus, support the idea that disinhibition is a common mechanism by which extrahippocampal structures modulate information flow through the hippocampus.

The neuroscience of spatial navigation: focus on behavior yields advances.

The development of the water maze as a laboratory approach to the study of spatial navigation has led to a large amount of research on the brain mechanisms of this ecologically important behavior. The procedural simplicity of this task belies its underlying complexity, which can complicate the interpretation of data obtained with the standard water maze procedure. In this review, recent experiments that used novel training procedures and detailed analyses of behavior are evaluated, together with related experiments, to clarify the brain mechanisms involved in this behavior. Pharmacological, lesion, and unit recording experiments demonstrate the existence of forebrain circuits for spatial navigation that are considerably more varied and extensive than was previously proposed, and involve various extrahippocampal structures. The use of novel and specialized procedures, together with a continued detailed focus on the behavior of animals in the maze, appears to be the most promising approach to understanding the mechanisms of spatial navigation.

A Study on the Invasive Hippocampal θ-Waves to the Cortex

Publisher Summary This chapter presents the experiments that are designed to investigate: (1) general features of the cortical “iso-hippocampal rhythm” (IHR), (2) the cortical distribution of the IHR, (3) conditions for the appearance of the cortical IHR, and (4) the cortical excitability during the IHR. The chapter uses the term IHR to denote the activity of extra-hippocampal structures which continues at least for 2 sec at frequencies of the θ of 4-7 c/sec and waxes and wanes in synchrony with the θ-waves of the hippocampus. The IHR activity could appear spontaneously, but it is much easier to induce it by the arousal stimulation. The IHRs presented in the chapter are mainly obtained by the electrical stimulation of the brain stem structures. One of the electrographic arousal states is characterized by cortical desynchronization and hippocampal synchronization. It is reported by Yoshii and associates that the cortical desynchronization sometimes contains a well-developed activity at the frequency-band corresponding to the hippocampal θwaves. The cortical activity with IHR is supposed to be moderately aroused and excludes the indifferent impulses coming to the cortex for inducing the correct performance in the conditioned and learned behavior.