Spontaneous Interictal Discharges Research Articles

Gamma-aminobutyric acidergic transmission underlies interictal epileptogenicity in pediatric focal cortical dysplasia.

Dysregulation of γ-aminobutyric acidergic (GABAergic) transmission has been reported in lesional acquired epilepsies (gliomas, hippocampal sclerosis). We investigated its involvement in a developmental disorder, human focal cortical dysplasia (FCD), focusing on chloride regulation driving GABAergic signals. In vitro recordings of 47 human cortical acute slices from 11 pediatric patients who received operations for FCD were performed on multielectrode arrays. GABAergic receptors and chloride regulators were pharmacologically modulated. Immunostaining for chloride cotransporter KCC2 and interneurons were performed on recorded slices to correlate electrophysiology and expression patterns. FCD slices retain intrinsic epileptogenicity. Thirty-six of 47 slices displayed spontaneous interictal discharges, along with a pattern specific to the histological subtypes. Ictal discharges were induced in proepileptic conditions in 6 of 8 slices in the areas generating spontaneous interictal discharges, with a transition to seizure involving the emergence of preictal discharges. Interictal discharges were sustained by GABAergic signaling, as a GABAA receptor blocker stopped them in 2 of 3 slices. Blockade of NKCC1 Cl- cotransporters further controlled interictal discharges in 9 of 12 cases, revealing a Cl- dysregulation affecting actions of GABA. Immunohistochemistry highlighted decreased expression and changes in KCC2 subcellular localization and a decrease in the number of GAD67-positive interneurons in regions generating interictal discharges. Altered chloride cotransporter expression and changes in interneuron density in FCD may lead to paradoxical depolarization of pyramidal cells. Spontaneous interictal discharges are consequently mediated by GABAergic signals, and targeting chloride regulation in neurons may be considered for the development of new antiepileptic drugs. Ann Neurol 2019; 1-14 ANN NEUROL 2019;85:204-217.

Interictal-like network activity and receptor expression in the epileptic human lateral amygdala

While the amygdala is considered to play a critical role in temporal lobe epilepsy, conclusions on underlying pathophysiological mechanisms have been derived largely from experimental animal studies. Therefore, the present study aimed to characterize synaptic network interactions, focusing on spontaneous interictal-like activity, and the expression profile of transmitter receptors in the human lateral amygdala in relation to temporal lobe epilepsy. Electrophysiological recordings, obtained intra-operatively in vivo in patients with medically intractable temporal lobe epilepsy, revealed the existence of interictal activity in amygdala and hippocampus. For in vitro analyses, slices were prepared from surgically resected specimens, and sections from individual specimens were used for electrophysiological recordings, receptor autoradiographic analyses and histological visualization of major amygdaloid nuclei for verification of recording sites. In the lateral amygdala, interictal-like activity appeared as spontaneous slow rhythmic field potentials at an average frequency of 0.39 Hz, which occurred at different sites with various degrees of synchronization in 33.3% of the tested slices. Pharmacological blockade of glutamate α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors, but not N-methyl-D-aspartate receptors, abolished interictal-like activity, while the γ-aminobutyric acid A-type receptor antagonist bicuculline resulted in a dampening of activity, followed by highly synchronous patterns of slow rhythmic activity during washout. Receptor autoradiographic analysis revealed significantly higher α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid, kainate, metabotropic glutamate type 2/3, muscarinic type 2 and adrenoceptor α(1) densities, whereas muscarinergic type 3 and serotonergic type 1A receptor densities were lower in the lateral amygdala from epileptic patients in comparison to autopsy controls. Concerning γ-aminobutyric acid A-type receptors, agonist binding was unaltered whereas antagonist binding sites were downregulated in the epileptic lateral amygdala, suggesting an altered high/low-affinity state ratio and concomitant reduced pool of total γ-aminobutyric acid A-type receptors. Together these data indicate an abnormal pattern of receptor densities and synaptic function in the lateral nucleus of the amygdala in epileptic patients, involving critical alterations in glutamate and γ-aminobutyric acid receptors, which may give rise to domains of spontaneous interictal discharges contributing to seizure activity in the amygdala.

Progestin receptors mediate progesterone suppression of epileptiform activity in tetanized hippocampal slices in vitro

Clinical and laboratory studies suggest that progesterone reduces epileptic seizure activity. The mechanisms underlying this effect are not known. The present study determined the effects of progesterone on extracellular evoked responses recorded in the CA1 field of hippocampal slices, as well as epileptiform responses recorded from tetanized slices. Slices were prepared from ovariectomized rats, with or without estrogen replacement. Hippocampal slices were superfused in vitro with one of the following treatments: progesterone with or without RU486 (a progesterone receptor antagonist); allopregnanolone (a progesterone metabolite that potentiates GABA action at GABA A receptors); RU5020 (a high-affinity progesterone receptor agonist); or cholesterol (control). In non-tetanized slices, a twofold increase in the excitatory postsynaptic field potential and population spike amplitude occurred during both cholesterol and progesterone superfusion. In contrast, under the same conditions, exposure to allopreganolone caused a 25% reduction in both field potential and population spike amplitude of evoked responses within 30 min of treatment. In tetanized slices, progesterone and RU5020, but not allopregnanolone or cholesterol, caused significant reductions in the field potential and population spike amplitude of evoked responses. Progesterone and RU5020 also significantly reduced the duration of tetanic stimulus-induced afterdischarges and the frequency of spontaneous interictal discharges. The effects of allopregnanolone were restricted to a reduction in the primary afterdischarge duration. Estrogen replacement slightly attenuated progesterone’s suppression of spontaneous discharges and depression of evoked responses. All responses to progesterone were blocked by prior or concurrent exposure to RU486. These data indicate that allopregnanolone suppresses evoked potentials in non-tetanized hippocampal slices, consistent with previous reports that this neurosteroid has marked anxiolytic and anticonvulsant effects. After tetanization, however, progesterone receptor-mediated responses become quantitatively more important as a mechanism for suppressing hippocampal electrical activity.

Kindling-induced changes in the EEG recorded during stimulation from the site of stimulation: II. Comparison between spontaneous and evoked potentials

We previously described how the EEG activity recorded during kindling-inducing tetanic stimulation, from the site of stimulation, consists of three major components. Here, we investigate the nature and role of component 1, an initial evoked potential, seen at the beginning of tetanus, and we compare it with the potential evoked by single-pulse stimulation and with the spontaneous interictal discharges seen after kindling in both the amygdala and hippocampus. The amplitude of component 1 increased during kindling, and the waveform, latency, and stimulus-response curve of component 1 were all similar to those of potentials evoked by single-pulse stimulation. Furthermore, in acute experiments, in which the amygdala was kindled by tetanic stimulation at 1-h intervals, the evoked potentials in the amygdala and pyriform cortex were markedly potentiated, whereas no obvious potential was recorded at the same latency from the ventral or dorsal hippocampus, subiculum, nor the prepyriform region. Thus, we conclude that, in the case of amygdala kindling, the nature of component 1 is a local direct and monosynaptic response involving the amygdala and pyriform cortex, and that the increased amplitude of component 1 after kindling may be related to long-term potentiation. Although the single-pulse stimulation produced late burst-like components in the amygdala and hippocampus after kindling, it produced neither EEG suppression, well-formed rhythmic synchronous bursts, nor epileptiform discharge, suggesting that component 1 alone is not sufficient for the subsequent events that may be essential to the seizure-triggering mechanism.

Anticonvulsant and proconvulsant actions of alpha- and beta-noradrenergic agonists on epileptiform activity in rat hippocampus in vitro.

The ability of l-norepinephrine to influence epileptiform activity was examined in an in vitro rat hippocampal slice preparation. Exogenously applied norepinephrine (NE) had anticonvulsant properties in that it slowed or stopped spontaneous interictal discharges which had been initiated by superfusion of slices with medium containing penicillin and elevated levels of potassium. This anticonvulsant property of NE was shared by the alpha receptor agonists 6-fluoro-norepinephrine, l-alpha-methyl-norepinephrine, and clonidine, but not by d-alpha-methyl-norepinephrine or l-phenylephrine. The beta receptor agonists 2-fluoro-norepinephrine and l-isoproterenol, on the other hand, were proconvulsant in that they increased interictal discharge rate. The alpha receptor antagonist phentolamine selectively blocked anticonvulsant responses, whereas the beta receptor antagonist timolol selectively blocked proconvulsant activity. These results suggest that activation of alpha or beta receptors has opposing inhibitory or excitatory effects respectively on epileptiform discharges in the rat hippocampus.

Endogenously released adenosine regulates excitability in the in vitro hippocampus.

The role of adenosine in regulating epileptiform discharge was studied in the in vitro hippocampal slice preparation. Exogenously applied adenosine appeared to have anticonvulsant properties (EC50 approximately 10 microM), in that it slowed or suppressed spontaneous interictal discharges. In addition, the release of adenosine (or related adenine nucleotides) from the slice preparation appeared to exert a tonic inhibitory influence on these types of discharges. This provides further evidence supporting a role for adenosine as an endogenous regulator of the excitability of the central nervous system, particularly in convulsive states.

Electrode placement and kindling-induced experiemental epilepsy

Electrical stimulation of the amygdala, caudate, hippocampus, or entorhinal cortex two or three times per day led to the development and progressive intensification of elicited motor seizures in rats. After a few hundred stimulations ( X = 348 ) the subjects in all four experimental groups displayed spontaneous motor seizures which continued to recur even after the regimen of stimulation had been curtailed. Although the development of spontaneous interictal discharges at the stimulation site preceded the initial manifestation of spontaneous motor seizures in all rats kindled by amygdaloid or hippocampal stimulation and in a few of the animals kindled through entorhinal electrodes, such discharges were not recorded in the caudate subjects.