Propagation Of Epileptiform Discharges Research Articles

In-vitro engineered human cerebral tissues mimic pathological circuit disturbances in 3D

In-vitro modeling of brain network disorders such as epilepsy remains a major challenge. A critical step is to develop an experimental approach that enables recapitulation of in-vivo-like three-dimensional functional complexity while allowing local modulation of the neuronal networks. Here, by promoting matrix-supported active cell reaggregation, we engineered multiregional cerebral tissues with intact 3D neuronal networks and functional interconnectivity characteristic of brain networks. Furthermore, using a multi-chambered tissue-culture chip, we show that our separated but interconnected cerebral tissues can mimic neuropathological signatures such as the propagation of epileptiform discharges.

Properties of the epileptiform activity in the cingulate cortex of a mouse model of LIS1 dysfunction

Dysfunction of the LIS1 gene causes lissencephaly, a drastic neurological disorder characterized by a deep disruption of the cortical structure. We aim to uncover alterations of the cortical neuronal networks related with the propagation of epileptiform activity in the Lis1/sLis1 mouse, a model lacking the LisH domain in heterozygosis. We did extracellular field-potential and intracellular recordings in brain slices of the anterior cingulate cortex (ACC) or the retrosplenial cortex (RSC) to study epileptiform activity evoked in the presence of bicuculline (10 µM), a blocker of GABAA receptors. The sensitivity to bicuculline of the generation of epileptiform discharges was similar in wild type (WT) and Lis1/sLis1 cortex (EC50 1.99 and 2.24 µM, respectively). In the Lis1/sLis1 cortex, we observed a decreased frequency of the oscillatory post-discharges of the epileptiform events; also, the propagation of epileptiform events along layer 2/3 was slower in the Lis1/sLis1 cortex (WT 47.69 ± 2.16 mm/s, n = 25; Lis1/sLis1 37.34 ± 2.43 mm/s, n = 15; p = 0.004). The intrinsic electrophysiological properties of layer 2/3 pyramidal neurons were similar in WT and Lis1/sLis1 cortex, but the frequency of the spontaneous EPSCs was lower and their peak amplitude higher in Lis1/sLis1 pyramidal neurons. Finally, the propagation of epileptiform activity was differently affected by AMPA receptor blockers: CNQX had a larger effect in both ACC and RSC while GYKI53655 had a larger effect only in the ACC in the WT and Lis1/sLis1 cortex. All these changes indicate that the dysfunction of the LIS1 gene causes abnormalities in the properties of epileptiform discharges and in their propagation along the layer 2/3 in the anterior cingulate cortex and in the restrosplenial cortex.

Altered activity and information flow in the default mode network of pilocarpine-induced epilepsy rats

Temporal lobe epilepsy (TLE) is considered a brain activity disorder that is likely related to allomnesia and conscious disturbance. In rodents, TLE epileptiform discharges can be triggered by systemic administration of a dose of pilocarpine. However, how these pilocarpine-induced epileptiform discharges generate and propagate through the whole brain has not been well studied. In this study, we sought to assess alterations of activity and information flow in the default mode network (DMN) during TLE epileptiform discharges in pilocarpine-induced TLE rats. We identified that in resting state, the rat DMN could be divided into three subnetworks that constituted a frequency-specific information flow loop. This frequency-specific loop converted into frequency-independent flow patterns during the generation and propagation of epileptiform discharges. Moreover, the activity of the theta (4–8 Hz) and alpha (8–13 Hz) bands in each DMN subnetwork exhibited completely different alterations during epileptiform discharges. Overall, our results demonstrated frequency-dependent alterations of both activity and information flow patterns within the DMN during epileptiform discharge. These results suggest that the independence and cooperation of different frequency bands may serve as an underlying mechanism for the generation and propagation of epileptiform discharges in the brain during SE in pilocarpine-treated rats.

Localization of Epileptogenic Zone With the Correction of Pathological Networks.

Patients with focal drug-resistant epilepsy are potential candidates for surgery. Stereo-electroencephalograph (SEEG) is often considered as the “gold standard” to identify the epileptogenic zone (EZ) that accounts for the onset and propagation of epileptiform discharges. However, visual analysis of SEEG still prevails in clinical practice. In addition, epilepsy is increasingly understood to be the result of network disorder, but the specific organization of the epileptic network is still unclear. Therefore, it is necessary to quantitatively localize the EZ and investigate the nature of epileptogenic networks. In this study, intracranial recordings from 10 patients were analyzed through adaptive directed transfer function, and the out-degree of effective network was selected as the principal indicator to localize the epileptogenic area. Furthermore, a coupled neuronal population model was used to qualitatively simulate electrical activity in the brain. By removing individual populations, virtual surgery adjusting the network organization could be performed. Results suggested that the accuracy and detection rate of the EZ localization were 82.86 and 85.29%, respectively. In addition, the same stage shared a relatively stable connectivity pattern, while the patterns changed with transition to different processes. Meanwhile, eight cases of simulations indicated that networks in the ictal stage were more likely to generate rhythmic spikes. This indicated the existence of epileptogenic networks, which could enhance local excitability and facilitate synchronization. The removal of the EZ could correct these pathological networks and reduce the amount of spikes by at least 75%. This might be one reason why accurate resection could reduce or even suppress seizures. This study provides novel insights into epilepsy and surgical treatments from the network perspective.

Electroencephalographic findings in patients with circumscribed thalamic lesions

IntroductionThalamo-cortical networks have mainly been studied in the generation of idiopathic (genetic) epilepsies. The purpose of this study was to analyze EEG patterns and the occurrence of focal (symptomatic) epileptic seizures in patients with acquired circumscribed thalamic lesions. Patients and methodsAmong 596 patients with thalamic lesions, we identified 47 patients in whom circumscribed thalamic lesions were detected by MRI and who underwent an EEG examination at the same stay at hospital. EEG findings were divided into normal findings, unspecific pathological changes and epileptiform discharges. The EEG findings were correlated to the localisation of the lesion within the thalamus and to the patientś symptoms. ResultsIn 32 patients (68%) pathological EEG findings were observed. They were heterogeneous and comprised regional and generalized slowing, triphasic waves, generalized periodic and regional epileptiform discharges. However, some characteristic findings were seen: Regional slowing was associated with ipsilateral thalamic lesions independent of the thalamic subarea, epileptiform discharges were related to lesions in the ipsilateral medial thalamus and periodic generalized discharges/triphasic waves with lesions in the anterior-ventromedial thalamus. Epileptic seizures were also more common in patients with medial thalamic lesions. Patients with regional epileptiform discharges responded to antiepileptic treatment whereas patients with triphasic waves and generalized periodic patterns did not. In some cases, it remained difficult to decide whether the thalamic lesion was the cause or consequence of epileptic activity. ConclusionPathological EEG findings are common in patients with acute and chronic thalamic lesions. EEG patterns associated with circumscribed thalamic lesions were influenced by the affected thalamic subregion. As in idiopathic generalized epilepsy, also in symptomatic epilepsy, the medial thalamus revealed to play a role in the generation of epileptiform discharges. In the patients with generalized periodic discharges and acute lesions in the ventral-anterior-medial thalamus, however, EEG changes were more likely caused by a disinhibition of cortico-thalamic networks than by a status epilepticus and thus risks and benefits of an aggressive antiepileptic treatment must be thoroughly balanced.

Temporal patterns of epileptiform discharges in genetic generalized epilepsies.

We sought to investigate the temporal patterns and sleep-wake cycle-related epileptiform discharges (EDs) in genetic generalized epilepsies (GGEs). We studied 24-hour ambulatory electroencephalography (EEG) recordings of patients with GGE, diagnosed and classified according to the International League against Epilepsy criteria. We manually coded the type of discharge, time of occurrence, duration, and arousal state of each ED. We employed mixed effects Poisson regression modeling to study the temporal distribution of epileptiform discharges. Additionally, we used multinomial regression analysis to explore the significance of the relationship between different states of arousal and types of epileptiform discharges. We analyzed 6923 EDs from 105 abnormal 24-hour EEGs. Mixed effects Poisson regression analysis demonstrated significant changes in ED counts across time blocks. This distribution was largely influenced by the state of arousal. Generalized fragments (duration<2s) and focal discharges were more frequent during non-REM sleep while paroxysms (duration≥2s) were more frequent in wakefulness. Overall, 67% of epileptiform discharges occurred in non-REM sleep and only 33% occurred in wakefulness. Twenty-four patients (23%) had ED exclusively in sleep. Epileptiform discharges peaked from 23:00 through 07:00h. There is a time-of-day dependency of ED with a significant influence exerted by the state of arousal. Our observations suggest that the generation of epileptiform discharges is not a random process but is the result of complex interactions among biological rhythms such as the sleep-wake cycle and the intrinsic circadian pacemaker. High density of ED in sleep suggests that 24-hour EEG recording with the capture of natural sleep may be more useful than routine EEG to diagnose GGE.

Transcranial Magnetic Stimulation Combined with EEG Reveals Covert States of Elevated Excitability in the Human Epileptic Brain.

Transcranial magnetic stimulation combined with electroencephalogram (TMS-EEG) can be used to explore the dynamical state of neuronal networks. In patients with epilepsy, TMS can induce epileptiform discharges (EDs) with a stochastic occurrence despite constant stimulation parameters. This observation raises the possibility that the pre-stimulation period contains multiple covert states of brain excitability some of which are associated with the generation of EDs. To investigate whether the interictal period contains "high excitability" states that upon brain stimulation produce EDs and can be differentiated from "low excitability" states producing normal appearing TMS-EEG responses. In a cohort of 25 patients with Genetic Generalized Epilepsies (GGE) we identified two subjects characterized by the intermittent development of TMS-induced EDs. The high-excitability in the pre-stimulation period was assessed using multiple measures of univariate time series analysis. Measures providing optimal discrimination were identified by feature selection techniques. The "high excitability" states emerged in multiple loci (indicating diffuse cortical hyperexcitability) and were clearly differentiated on the basis of 14 measures from "low excitability" states (accuracy = 0.7). In GGE, the interictal period contains multiple, quasi-stable covert states of excitability a class of which is associated with the generation of TMS-induced EDs. The relevance of these findings to theoretical models of ictogenesis is discussed.

Investigation of The Initiation Site and Propagation of Epileptiform Discharges in Hippocampal Slices Using Microelectrode Array

Understanding the initiation site and propagation of epileptiform discharges are of important significance for investigating the mechanisms of epilepsy and thereby for the clinically remedy.In order to solve the above problems,epileptiform discharges induced by low-Mg2+artificial cerebrospinal fluid(ACSF) in Sprague-Dawley(SD) rat hippocampal slices were recorded by microelectrode array.Two components of epileptiform discharges:field potentials and multiple unit activity(MUA) were analyzed.Firstly,the onset time of field potentials in stratum pyramidale was calculated and compared to locate the initiation site and propagation of epileptiform discharges.Then cross-correlation analysis was applied to spike trains of MUA.Time delays obtained from cross-correlation function further confirmed the initiation site and propagation of epileptiform discharges in the whole hippocampal slice.The results revealed that epileptiform discharges in CA3 had larger amplitudes and longer duration than that in CA1,which indicated more excitability in CA3.Field potentials as well as MUA in CA3b occurred earlier compared with the synchronous signals in CA3c and CA1.The time delays between the onset and its following areas were positive relative to the distances between them.The result demonstrated that in low-Mg2+ACSF-perfused SD rat hippocampal slice,epileptiform discharges originated from CA3b and propagated to CA3c and CA1 region respectively.

Association Study of TRPC4 as a Candidate Gene for Generalized Epilepsy with Photosensitivity

Photoparoxysmal response (PPR) is characterized by abnormal visual sensitivity of the brain to photic stimulation. Frequently associated with idiopathic generalized epilepsies (IGEs), it might be an endophenotype for cortical excitability. Transient receptor potential cation (TRPC) channels are involved in the generation of epileptiform discharges, and TRPC4 constitutes the main TRPC channel in the central nervous system. The present study investigated an association of PPR with sequence variations of the TRPC4 gene. Thirty-five single nucleotide polymorphisms (SNP) within TRPC4 were genotyped in 273 PPR probands and 599 population controls. Association analyses were performed for the broad PPR endophenotype (PPR types I-IV; n = 273), a narrow model of affectedness (PPR types III and IV; n = 214) and PPR associated with IGE (PPR/IGE; n = 106) for each SNP and for corresponding haplotypes. Association was found between the intron 5 SNP rs10507456 and PPR/IGE both for single markers (P = 0.005) and haplotype level (P = 0.01). Three additional SNPs (rs1535775, rs10161932 and rs7338118) within the same haplotype block were associated with PPR/IGE at P < 0.05 (uncorrected) as well as two more markers (rs10507457, rs7329459) located in intron 3. Again, the corresponding haplotype also showed association with PPR/IGE. Results were not significant following correction for multiple comparisons by permutation analysis for single markers and Bonferroni-Holm for haplotypes. No association was found between variants in TRPC4 and other phenotypes. Our results showed a trend toward association of TRPC4 variants and PPR/IGE. Further studies including larger samples of photosensitive probands are required to clarify the relevance of TRPC4 for PPR and IGE.

Carisbamate prevents the development and expression of spontaneous recurrent epileptiform discharges and is neuroprotective in cultured hippocampal neurons

Although great advances have been made in the development of treatments for epilepsy, acquired epilepsy following brain injury still comprises approximately 50% of all the cases of epilepsy. Thus, development of drugs that would prevent or decrease the onset of epilepsy following brain injury represents an important area of research. Here, we investigated effects of carisbamate (RWJ 333369) on the development and expression of spontaneous recurrent epileptiform discharges (SREDs) and its neuroprotective potential in cultured hippocampal neurons. This model utilizes 3 h of low Mg(2+) treatment to mimic status epilepticus (SE-like) injury in vitro. Following the injury, networks of neurons manifest synchronized SREDs for their life in culture. Neuronal cultures were treated with carisbamate (200 microM) for 12 h immediately after the SE-like injury. The drug was then removed and neurons were patch clamped 24 h following drug washout. Treatment with carisbamate after neuronal injury prevented the development and expression of epileptiform discharges. In the few neurons that displayed SREDs following carisbamate treatment, there was a significant reduction in SRED frequency and duration. In contrast, phenytoin and phenobarbital, when used in place of carisbamate, did not prevent the development and expression of SREDs. Carisbamate was also effective in preventing neuronal death when administered after SE-like injury. Carisbamate prevents the development and generation of epileptiform discharges and is neuroprotective when administered following SE-like injury in vitro and may offer a novel treatment to prevent the development of epileptiform discharges following brain injuries.

Modelling large motion events in fMRI studies of patients with epilepsy

EEG-correlated fMRI can provide localisation information on the generators of epileptiform discharges in patients with focal epilepsy. To increase the technique's clinical potential, it is important to consider ways of optimising the yield of each experiment while minimizing the risk of false-positive activation. Head motion can lead to severe image degradation and result in false-positive activation and is usually worse in patients than in healthy subjects. We performed general linear model fMRI data analysis on simultaneous EEG-fMRI data acquired in 34 cases with focal epilepsy. Signal changes associated with large inter-scan motion events (head jerks) were modelled using modified design matrices that include 'scan nulling' regressors. We evaluated the efficacy of this approach by mapping the proportion of the brain for which F-tests across the additional regressors were significant. In 95% of cases, there was a significant effect of motion in 50% of the brain or greater; for the scan nulling effect, the proportion was 36%; this effect was predominantly in the neocortex. We conclude that careful consideration of the motion-related effects in fMRI studies of patients with epilepsy is essential and that the proposed approach can be effective.

Wavelet-Crosscorrelation Analysis: Non-Stationary Analysis of Neurophysiological Signals

Wavelet-crosscorrelation analysis is a new application of wavelet analysis used to show the propagation of epileptiform discharges and to localize the corresponding lesions. We have shown previously that this analysis can help predict brain conditions statistically (Mizuno-Matsumoto et al. 2002). Our objective was to assess whether wavelet-crosscorrelation analysis reveals the initiation and propagation of epileptiform activity in human patients. The data obtained from three patients with simple partial seizures (SPS) using whole-head magnetoencephalography (MEG) were analyzed by the wavelet-crosscorrelation method. Wavelet-crosscorrelation coefficients (WCC), the coherent structure of each possible pair of signals from 64 MEG channels forvarious periods, and the time lag (TL) in two related signals, were ascertained. We clearly demonstrated both localization of the irritative zone and propagation of the epileptiform discharges. Wavelet-crosscorrelation analysis can help reveal and visualize the dynamic changes of brain conditions. The method of this analysis can compensate for other existing methods for the analysis of MEG, electroencephalography (EEG) or Elecotrocorticography (ECoG). Our proposed method suggests that revealing and visualizing the dynamic changes of brain conditions can help clinicians and even patients themselves better understand such conditions.

Callosal role in generation of epileptiform discharges: quantitative analysis of EEGs recorded in patients undergoing corpus callosotomy.

Corpus callosotomy tends to decrease seizure frequency and severity rather than transforming the seizure pattern from a generalized form into a lateralized or a partial one. The finding implies that bisection of the corpus callosum (CC) disrupts the epileptogenicity itself. In order to gain further insight into the possible role played by the CC in intractable generalized epilepsy, quantitative analyses of long-term EEGs were performed. Analyses were made of epileptiform discharges contained in the pre- and postoperative long-term EEGs in 7 patients who had an anterior callosotomy for intractable epilepsy. The duration, number, and amplitude of all epileptiform burst activities were measured and statistically analyzed. After callosotomy, the total number of epileptiform burst activities, mean duration, and the total number of spike discharges decreased significantly. The two hemispheres could be divided into dominant and non-dominant ones as to the quantity of the residual epileptiform discharges. Corpus callosotomy unevenly reduced preoperative epileptiform discharges in both hemispheres, suggesting a facilitatory role played by the callosal neurons that enables the asymmetrical epileptogenic susceptible state of the two hemispheres to develop bisynchronous and bisymmetrical epileptiform discharges. Corpus callosotomy decreased the quantity of the epileptiform discharges, suggesting the transhemispheric facilitation of seizure mechanisms.

Distribution of ion channels in trigeminal ganglion neuron of rat

Dorsal root ganglia (DRG) neurons transmit various sensory information such as touch, pressure, pain and temperature from the peripheral region to the central nervous system. Similarly, a pair of trigeminal root ganglia (TRG) is responsible for the sensory inputs from the oromaxillofacial region to which the trigeminal nerve is innervated. The sensory neurons of mammalian DRG are known to express voltage-dependent ion channels: at least two types of sodium currents (INa) which are distinguished by their sensitivity to TTX (Bossu and Feltz 1984; Caffrey et al. 1992; Elliott and Elliott 1993; Fedulova et al. 1991; Kostyuk et al. 1981, McLean et al. 1988; Ogata and Tatebayashi 1993; Ogata and Tatebayashi 1992 a, b; Roy and Narahashi 1992; Schwartz et al. 1990). In addition, the distribution of these ion channels in DRG neurons, according to the size of cells and developmental stages, has been widely studied to elucidate the developmental and functional role of the each type of ion channels in the modulation of specific sensory modalities (Roy and Narahashi 1992; Schwartz et al. 1990). Voltage-activated calcium channels have been implicated in a number of important neuronal functions, including secretion of neurotransmitters and hormones (Perney et al. 1986; Hirning et al. 1988; Holz et al. 1988), generation of epileptiform discharges (Wong and Prince 1978) and activation of intracellular enzymes (Nishizuka 1984). Ca influx through voltage-activated calcium channels might also contribute to neurodegeneration associated with ischaemia (Choi 1988). For Distribution of ion channels in trigeminal ganglion neuron of rat

Landau–Kleffner Syndrome: Localization of Epileptogenic Lesion Using Wavelet– Cross-Correlation Analysis

Magnetoencephalographic findings in a 6-year-old patient suffering from acquired aphasia with convulsive disorder (Landau–Kleffner Syndrome, LKS) are presented. The data were analyzed using wavelet–cross-correlation analysis, a nonstationary analysis method developed to analyze the localization of an epileptogenic lesion and the propagation of epileptiform discharges. The results indicate that LKS might be a disorder of the primary temporal cortex, and that the auditory neural network may function as the circuit for the epileptic discharge propagation.

Origin and propagation of spontaneous electrographic sharp waves in the in vitro turtle brain: a model of neuronal synchronization

Objectives: Neuronal synchronization is a basic feature in the generation of epileptiform discharges. Spontaneous large sharp waves (LSWs) can be recorded in the turtle brain in vitro, indicating the synchronous activation of large neuronal populations. The aim of this study was to analyze the spatial and temporal distribution of LSWs within the brain; the participation of glutamate in LSWs generation was also investigated.Methods: Extracellular field potentials were recorded in vivo (n=4) and in vitro (n=36). LSWs were recorded from cerebral cortex, optic tectum, and thalamus.Results: LSWs were recorded from cerebral cortex, optic tectum and thalamus. No LSWs were observed in cerebellum and brain stem. In some experiments, LSWs could be recorded only from medial cortex. Latency studies demonstrated that, within each hemisphere, medial cortex led the generation of LSWs; in addition, isolated medial cortex could sustain LSWs. Intracortical laminar field potentials in medial cortex indicated that LSWs generate mainly in the molecular layer, probably at pyramidal cell dendrites. Pharmacological experiments demonstrated that NMDA and non-NMDA glutamate receptors are involved in LWSs generation.Conclusions: These results suggest that turtle medial cortex is the pacemaker area for LSWs generation and it can be a useful model to study cellular and circuital mechanisms of neuronal synchronization.

Epileptiform activity generated by endogenous acetylcholine during blockade of GABAergic inhibition in immature and adult rat hippocampus

We tested the effects of the acetylcholinesterase inhibitor eserine (10 μM), an indicator of the activity of endogenous ACh, on the generation of epileptiform discharges during blockade of inhibitory GABA A-mediated potentials by bicuculline (10 μM), in the CA3 area of hippocampal slices from postnatal days 4–20 (P4–P20) immature and adult rats. Eserine provoked or significantly increased the frequency of spontaneous synchronous epileptiform discharges, in 6/22 (27%) P4–P10 slices, 34/35 P11–P20 slices and 18/18 adult slices, an epileptogenic effect. In immature slices, spontaneous discharges showed a stable frequency throughout perfusion with eserine, while in 5/11 adult slices an initial fast frequency was followed by a slower steady-state one. The cholinergic agonist carbachol (CCh, 25 μM) provoked only transient or no spontaneous synchronous discharges in adult slices ( n=8), thus suggesting that massive activation of cholinergic receptors may lead to suppression of epileptiform activity in adult brain. Stimulus-induced excitatory CA3 responses, were depressed by eserine in approximately half of 20 P4–P10, 45 P11–P20 and 11 adult slices. The depression consisted of a decrease in the amplitude, duration, and number of population spikes of the field potentials by about 30%, a minor neuroprotective effect, which did not change with maturation. The different developmental profiles of the epileptogenic and neuroprotective effects of endogenous ACh suggest that they are mediated by different mechanisms. These experiments demonstrate that, endogenous ACh is sufficient to induce epileptogenesis during a decrease or failure of GABAergic inhibition, in both ≥P10 immature and in adult hippocampus. We therefore suggest that clinical or behavioral conditions which raise the concentration of endogenous ACh may lower the threshold to seizures.

Reduced signal complexity of intracellular recordings: a precursor for epileptiform activity?

Recent studies have shown that time windowed extraction of nonlinear parameters like an effective correlation dimension from intracranially recorded EEG of epileptic patients often allows to detect and identify an unequivocal “pre-ictal phase” preceding an epileptic seizure. In another study, however, such an anticipation could not be made. These conflicting findings may indicate that observed changes in nonlinear parameters probably depend on the type of elementary mechanisms underlying epileptic processes and/or the spatial distribution of neurons primarily involved in generation of epileptiform discharges. To test the existence of such dependencies, the transition from normal to epileptiform activity (EA) of CA3-neurons in hippocampal slices was analyzed in four epilepsy models, using a time windowed computation of an effective correlation dimension. Indeed, in xanthine and penicillin models, signal complexity in intracellular recordings was reduced before manifestation of paroxysmal depolarization shifts (PDS), whereas a preceding loss of complexity was missing in low-magnesium and veratridine models. These findings indicate that interictal-like EA is predictable only in some epilepsy models.

Visualization of epileptogenic phenomena using cross-correlation analysis: localization of epileptic foci and propagation of epileptiform discharges.

The main objectives of the preoperative evaluation of a patient with medically intractable epileptic seizures are localization of the foci and propagation of the epileptiform discharges. Electrocorticographic (ECoG) data of intractable focal epilepsy were analyzed using an AR model, wavelet analysis, and cross-correlation analysis. In order to derive the time-shifts, the cross correlations of the epileptiform discharges were calculated between electrodes for every unit of time. Further analyses were made by means of a set of contour maps of the time-shifts and sequential two- and three-dimensional visualizations of the time-shift maps in order to localize the epileptic foci and study their propagation process. Two types of foci and propagation were revealed in the results. In the first type, epileptiform discharges were generated at localized focal sites and spread quickly to other sites. In the second type, the foci of epileptiform discharges, which appeared soon after the former bursts, were localized at more than one site, and the discharges tended to spread more slowly. The findings suggest that epileptic phenomena can be caused by at least two kinds of mechanisms in one patient: in the former, the propagation might be mediated through synaptic projections, while in the latter, the extracellular diffusion of an excitatory factor might play an important role. In addition, our newly developed visualization technique for the localization of epileptic foci and the propagation of epileptiform discharges should prove useful in the study of epileptogenesis etiology.

Laminar organization of epileptiform discharges in the rat entorhinal cortex in vitro.

1. Interictal and ictal epileptiform discharges induced by 4-aminopyridine (4AP, 50 microM) were studied in the rat lateral entorhinal cortex with field potential and intracellular recordings in an in vitro slice preparation. Both types of discharge disappeared in layer II, but continued to occur in layers IV-VI after a knife cut separation was made at approximately 600 micro(m) from the pia (n = 4 slices). 2. Interictal depolarizations recorded in layer IV-VI cells (amplitude, 29.4 +/- 8.6 mV; duration, 386 +/- 177.4 ms, means +/- s.d.; n = 17) were capped by action potential bursts, while smaller interictal depolarizations in layer II cells (amplitude, 11.7 +/- 5.8 mV; duration, 192.6 +/- 47.9 ms; n = 10) were associated with single action potentials and were terminated by a hyperpolarization. Ictal discharges were initiated by an interictal discharge; they were characterized by a depolarization of 31.5 +/- 6.2 mV (n = 12) in layer IV-VI and 11.6 +/- 3.5 mV (n = 7) in layer II neurones. 3. Slow, presumptive Ca2+-mediated spikes occurred in layer II (n = 4) and IV-VI (n = 6) cells loaded with the Na+ channel blocker QX-314 (50 mM). These events were synchronized with population spikes during interictal and ictal discharges, and were abolished by Ni2+ (1 mM, n = 4 cells) along with the 4AP-induced synchronous activity. 4. The N-methyl-D-aspartate (NMDA) receptor antagonist 3, 3-(2-carboxypiperazine-4-yl)-propyl-1-phosphonate (CPP, 10 microM) abolished ictal discharges and reduced interictal depolarizations in layer IV-VI neurones (n = 4). The non-NMDA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 microM) abolished both interictal and ictal activity (n = 4 cells). 5. These findings provide evidence for a role played by NMDA-mediated mechanisms in the generation of epileptiform discharges in the entorhinal cortex. Lack of an NMDA-mediated component along with presence of inhibition in layer II neurones results in attenuation of epileptiform activity at this site. Moreover Ca2+-mediated spikes may contribute to the appearance of epileptiform discharges in this model.