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M Avoli

Publications and source records attributed to M Avoli.

At least 55 records · Page 3Linked to original sources
PubMed1997

CA3-driven hippocampal-entorhinal loop controls rather than sustains in vitro limbic seizures.

Continuous application of 4-aminopyridine (4-AP, 50 microM) to combined slices of hippocampus-entorhinal cortex obtained from adult mice induces (1) interictal discharges that initiate in the CA3 area and propagate via the hippocampal regions CA1 and subiculum to the entorhinal cortex and return to the hippocampus through the dentate gyrus; and (2) ictal discharges that originate in the entorhinal cortex and propagate via the dentate gyrus to the hippocampus proper. Ictal discharges disappear over time, whereas synchronous interictal discharges continue to occur throughout the experiment. Lesioning the Schaffer collaterals abolishes interictal discharges in CA1, entorhinal cortex, and dentate gyrus and discloses entorhinal ictal discharges that propagate, via the dentate gyrus, to the CA3 subfield. Interictal discharges originating in CA3 also prevent the occurrence of ictal events generated in the entorhinal cortex during application of Mg2+-free medium. In both models, ictal discharge generation recorded in the entorhinal cortex after Schaffer collateral cut is prevented by mimicking CA3 neuronal activity through rhythmic electrical stimulation (0.25-1.5 Hz) of the CA1 hippocampal output region. Our findings demonstrate that interictal discharges of hippocampal origin control the expression of ictal epileptiform activity in the entorhinal cortex. Sectioning the Schaffer collaterals may model the chronic epileptic condition in which cell damage in the CA3 subfield results in loss of CA3 control over the entorhinal cortex. Hence, we propose that the functional integrity of hippocampal output neurons may represent a critical control point in temporal lobe epileptogenesis.

4-Aminopyridine↗
PubMed1997

Topiramate attenuates voltage-gated sodium currents in rat cerebellar granule cells.

Whole-cell, voltage-clamp recordings were made from rat cerebellar granule cells in culture under experimental conditions designed to study voltage-gated Na+ currents that were elicited by depolarizing commands from a holding potential of -60 mV up to +20 mV. These tetrodotoxin-sensitive inward currents were reduced in a dose-related manner by bath application of the structurally novel, anticonvulsant drug topiramate (10-1000 microM; n = 16). Dose-response analysis of this effect revealed an IC50 of 48.9 microM. Topiramate also made the steady-state inactivation curve of this current shift toward more negative values (midpoint of the inactivation curve -46.9 mV under control conditions and -56.5 mV during topiramate application; n = 5). We propose that these effects may contribute to control the sustained depolarizations with repetitive firing of action potentials that occur within neuronal networks during seizure activity. Therefore they may represent a mechanism of action for this novel anticonvulsant drug.

Animals↗
PubMed1997

In vitro electrophysiology of rat subicular bursting neurons.

Intracellular recordings were used to study the electrophysiological properties of rat subicular neurons in a brain slice preparation in vitro. Cells were classified as bursting neurons (n = 102) based on the firing pattern induced by depolarizing current pulses. The bursting response recorded at resting membrane potential (-66.1 +/- 6.2 mV, mean +/- SD n = 94) was made up of a cluster of fast action potentials riding on a slow depolarization and was followed by an afterhyperpolarization. Tonic firing occurred at a membrane potential of approximately -55 mV. A burst also occurred upon termination of a hyperpolarizing current pulse. Tetrodotoxin (TTX, 1 microM) blocked the burst and decreased or abolished the underlying slow depolarization. These effects were not induced by the concomitant application of the Ca2+ channel blockers Co2+ (2 mM) and Cd2+ (1 mM). Subicular bursting neurons displayed voltage- and time-dependent inward rectifications of the membrane during depolarizing and hyperpolarizing current pulses. The inward rectification in the depolarizing direction was abolished by TTX, while that in the hyperpolarizing direction was blocked by extracellular Cs+ (3 mM), but not modified by Ba2+ (0.5-1 mM), TTX, or Co2+ and Cd2+. Tetraethylammonium (10 mM)-sensitive, outward rectification became apparent in the presence of TTX. These results suggest that neurons in the rat subiculum can display voltage-dependent bursts of action potentials as well as membrane rectification in the depolarizing and hyperpolarizing directions. These results also indicate that activation of a voltage-gated Na+ conductance may be instrumental in the initiation of bursting activity.

Action Potentials↗
PubMed1997

Repetitive firing and oscillatory activity of pyramidal-like bursting neurons in the rat subiculum.

Electrophysiological characterization of neurons within the rat subiculum was carried out with intracellular recordings in an in vitro slice preparation. Subicular neurons responded to threshold pulses of depolarizing current delivered at a resting membrane potential (RMP) of 45.7+/-5.8 mV (mean+/-SD, n=85) with an initial burst of three to five fast action potentials that rode on a depolarizing envelope and was terminated by an afterhyperpolarization (burst AHP) (duration 113+/-35 ms; peak amplitude 2.7+/-0.6 mV, n=10). Tonic firing replaced the bursting mode at membrane potential less negative than -55 mV. Suprathreshold depolarizing pulses evoked at RMP both an initial burst and successive tonic firing. Intracellular staining with biocytin showed morphological features typical of pyramidal cells (n=8). The relationship between frequency of repetitive firing and injected current (f-I) revealed that the burst firing frequency (250-300 Hz) was only slightly influenced by the amount of injected current. By contrast, the f-I curve of the tonic firing phase depended upon current intensity: it displayed an initial segment that increased at first linearly and then turned into a plateau for both the early and the late inter-spike intervals. The frequency of the tonic firing declined only slightly with time, thus suggesting a lack of adaptation. During tonic firing, each single action potential was followed by a fast AHP and a depolarizing afterpotential. Termination of repetitive firing was followed by an AHP (spike-train AHP; duration 223+/-101 ms, peak amplitude 5.6+/-2.4 mV, n=17). Fast spike-train and burst AHPs were reduced by bath application of the Ca2+-channel blockers Co2+ (2 mM) and Cd2+ (1 mM) (n=8), thus suggesting the participation of Ca2+-dependent K+ conductances in these AHPs. Subicular bursting neurons generated persistent, subthreshold voltage oscillations at 5.3+/-1 Hz (n=20) during steady depolarization positive to -60 mV; at values positive to -55 mV, the oscillatory activity could trigger clusters of single action potentials with a periodicity of 0.9-2 Hz. Oscillations were not prevented by application of excitatory amino acid receptor and GABA(A) receptor antagonists (n=5), Ca2+-channel blockers (n=5), or Cs+ (3 mM; n=4), but were abolished by the Na+-channel blocker tetrodotoxin (1 microM; n=6). Our findings demonstrate that pyramidal-like subicular neurons generate both bursting and non-adapting tonic firing, depending upon their membrane potential. These neurons also display oscillatory activity in the range of theta frequency that depends on the activation of a voltage-gated Na+ conductance. These electrophysiological properties may play a role in the process of signals arising from the hippocampal formation before being funnelled towards other limbic structures.

Action Potentials↗
PubMed1997

Pilocarpine-induced seizures are accompanied by a transient elevation in the messenger RNA expression of the prohormone convertase PC1 in rat hippocampus: comparison with nerve growth factor and brain-derived neurotrophic factor expression.

Several prohormone convertases that are involved in the posttranslational processing of precursor proteins, including neuropetides, hormones and neurotrophic factors, are produced in the central nervous system. These include enzymes named furin, PC1, PC2, PC5 and PACE4. To understand better the potential role played by prohormone convertases in the central nervous system we studied the expression of their messenger RNAs in the hippocampus of rats with pilocarpine-induced seizures. Moreover, we compared their expression patterns with those of neurotrophins such as nerve growth factor and brain-derived neurotrophic factor, which are up-regulated in the hippocampus during seizures. Pilocarpine (380 mg/kg, i.p.) induced seizure activity that appeared within the first hour and persisted for approximately 8 h. In situ hybridization showed transient increases in messenger RNA for nerve growth factor and brain-derived neurotrophic factor that peaked at 120 min in the hippocampus. Among the convertases studied, only PC1 messenger RNA displayed up-regulation, with temporal and topographic features comparable to those of nerve growth factor and brain-derived neurotrophic factor messenger RNA. The expression of furin, PC2 and PC5 messenger RNA changed little, while PACE4 was not expressed at all, both before and after pilocarpine administration. The highest increase in PC1 messenger RNA expression was found in granule cells of the dentate gyrus and, to a lesser extent, in the pyramidal layer of CA1 and CA3 subfields. Thus, in the rat hippocampus, the epileptiform activity induced by pilocarpine mediates a co-ordinated expression of messenger RNAs for PC1, nerve growth factor and brain-derived neurotrophic factor. Our findings suggest the involvement of PC1 in the processing of precursor proteins during seizure activity.

Animals↗
PubMed1997

Lamotrigine reduces voltage-gated sodium currents in rat central neurons in culture.

PURPOSE: To study the mechanism or mechanisms of action of lamotrigine (LTG) and, in particular, to establish its effects on the function of NA+ channels in mammalian central neurons. METHODS: Rat cerebellar granule cells in culture were subjected to the whole-cell mode of voltage clamping under experimental conditions designed to study voltage-gated Na+ currents. RESULTS: Extracellular application of LTG (10-500 microM, n = 21) decreased in a dose-related manner a tetrodotoxin-sensitive inward current that was elicited by depolarizing commands (from -80 to +20 mV). The peak amplitude of this Na(+)-mediated current was diminished by 38.8 +/- 12.2% (mean +/- SD, n = 6) during application of 100 microM LTG, and the dose-response curve of this effect indicated an IC50 of 145 microM. The reduction in the inward currents produced by LTG was not associate with any significant change in the current decay, whereas the voltage dependency of the steady-state inactivation shifted toward more negative values (midpoint of the inactivation curve: -47.5 and -59.0 mV under control conditions and during application of 100 microM LTG, respectively, n = 4). CONCLUSIONS: Our findings indicate that LTG reduces the amplitude of voltage-gated Na+ inward current in rat cerebellar granule cells and induces a negative shift of the steady-state inactivation curve. Both mechanisms may be instrumental in controlling the repetitive firing of action potentials (AP) that occurs in neuronal networks during seizure activity.

Animals↗
PubMed1997

Function and organization in dysgenic cortex. Case report.

Cerebral dysgenesis is a subject of interest because of its relationship to cerebral development and dysfunction and to epilepsy. The authors present a detailed study of a 16-year-old boy who underwent surgery for a severe seizure disorder. This patient had dysgenesis of the right hemisphere, which was composed of a giant central frontoparietal nodular gray matter heterotopia with overlying large islands of cortical dysplasia around a displaced central fissure. Exceptional insight into the function, biochemistry, electrophysiology, and histological structure of this lesion was obtained from neurological studies that revealed complementary information: magnetic resonance (MR) imaging, [18]fluoro-2-deoxy-D-glucose positron emission tomography (PET), functional PET scanning, proton MR spectroscopic (1H-MRS) imaging, intraoperative cortical mapping and electrocorticography, in vitro electrophysiology, and immunocytochemistry. These studies demonstrated compensatory cortical reorganization and showed that large areas of heterotopia and cortical dysplasia in the central area may retain normal motor and sensory function despite strikingly altered cytoarchitectonic organization and neuronal metabolism. Such lesions necessitate appropriate functional imaging studies prior to surgery and cortical mapping to avoid creating neurological deficits. Integrated studies, such as PET, 1H-MRS imaging, cortical mapping, immunocytochemistry, and electrophysiology may provide information on the function of developmental disorders of cerebral organization.

Adolescent↗
PubMed1997

Functional and pharmacological properties of GABA-mediated inhibition in the human neocortex.

This paper describes some functional and pharmacological properties of GABA-mediated mechanisms in the human neocortex maintained in vitro in a slice preparation. Neocortical neurons recorded intracellularly under normal conditions generate stimulus-induced and spontaneous potentials that are mediated by the activation of postsynaptic GABAA and GABAB receptor subtypes. As reported in other species, pharmacological blockade of the GABAA receptor makes epileptiform bursts appear in response to extracellular focal stimuli, thus indicating that inhibition mediated through the activation of the GABAA receptor exerts an important role in controlling neuronal excitability in the human neocortex. Spontaneous, prolonged epileptiform discharge are recorded when slices are bathed in Mg(2+)-free medium. Under these experimental conditions GABAA receptor mediated potentials occur between epileptiform events; moreover their rate of occurrence decreases shortly before the onset of each discharge. Blockade of GABAA receptor mediated potentials during application of Mg(2+)-free medium (i) prolongs the epileptiform discharges, (ii) increases the amplitude of their field potential DC shifts, and (iii) augments the concomitant decreases in [Ca2+]0 and increases in [K+]0. These findings indicate therefore that GABAA receptor mediated inhibitory potentials are operant during Mg(2+)-free epileptiform activity, and modulate the occurrence of epileptiform discharges. Moreover, they may also play a role in controlling the changes in [Ca2+]0 and [K+]0 that accompany each epileptiform event.

Bicuculline↗
PubMed1996

Excitatory effects induced by carbachol on bursting neurons of the rat subiculum.

Conventional intracellular recordings were made from neurons of the rat subiculum in an in vitro slice preparation. Intracellular pulses of depolarizing current (duration, 10-120 ms) delivered at a resting membrane potential of -62.2 +/- 7.7 mV (mean +/- SD, n = 14) induced bursts of 3-5 fast, action potentials riding on a slow depolarization. The burst was terminated by an afterhyperpolarization (burst AHP) that lasted 117 +/- 26 ms and reached peak amplitude of 5.1 +/- 1.8 mV (n = 8). Bath application of the cholinergic agonist carbachol (CCh; 30-100 microM; n = 20) in the presence of ionotropic excitatory amino acid receptor antagonists induced a steady depolarization (4.6 +/- 2.7 mV) of the membrane potential, and a small increase in input resistance. Action potential bursts continued to occur in response to intracellular depolarizing pulses during CCh application. However, this cholinergic agonist reduced and eventually blocked the burst AHP, which was replaced by action potentials firing. In the presence of CCh (> 70 microM; n = 9) the burst response, was followed by a depolarizing plateau potential (PP) that outlasted the intracellular depolarizing pulse by 731 +/- 386 ms (range 160-1900 ms), and could trigger repetitive action potential firing at 35-116 Hz. The effects induced by CCh were reversed by bath application of the muscarinic antagonist atropine (0.5-1 microM; n = 4). Our findings demonstrate that CCh exerts in the rat subiculum an excitatory action that is dependent upon muscarinic receptor stimulation. This cholinergic mechanism may play a physiological role in the subicular processing of signals arising from the hippocampus proper, and may also contribute to the generation of sustained epileptiform discharges induced in the limbic system by cholinergic agents.

Animals↗
PubMed1996

Synchronization of rat hippocampal neurons in the absence of excitatory amino acid-mediated transmission.

Extracellular and intracellular recordings and measurements of extracellular K+ concentration ([K+]o) were performed in the adult rat hippocampus in an in vitro slice preparation. Excitatory amino acid receptor antagonists, as well as the K(+)-channel blockers 4-aminopyridine (4AP, 50 microM) and/or tetraethylammonium (TEA, 5 mM), were added to the bath. Synchronous, negative-going field potentials were recorded in the CA3 stratum radiatum during application of 4AP and excitatory amino acid receptor antagonists. Each of these events was associated with an intracellular long-lasting depolarization and a concomitant rise in [K+]o that attained peak values of 4.3 +/- 0.1 mM (mean +/- S.E.M., n = 6 slices) and lasted 29 +/- 3 s. These field potentials were still recorded in CA3 stratum radiatum after addition of TEA. Under these conditions, prolonged field potentials (40.2 +/- 4.5 s, n = 18) characterized by a prominent positive component; discharge of population spikes also occurred. [K+]o increases associated with these prolonged field-potential discharges had a considerable variability in magnitude (peak value = 3.8-14.1 mM, 6.1 +/- 0.7 mM, n = 5) and duration (14-210 s; 48 +/- 13 s, n = 5). In 8% of the cases spreading depression-like episodes were observed. [K+]o increases during spreading depression-like episodes attained peak values of 11-27 mM (22.8 +/- 0.2 mM, n = 2) and had a duration of 160-396 s (244 +/- 29 s, n = 2). All types of synchronous activity were abolished by the GABAA-receptor antagonist bicuculline methiodide (10 microM) (n = 11). A similar effect was obtained by applying Ca(2+)-free/high-Mg2+ medium (n = 5). Simultaneous field-potential recordings in CA3, CA1, dentate area and subiculum demonstrated that negative-going potentials and prolonged field-potential discharges occurred in all areas in a synchronous fashion. Spreading depression-like episodes were more frequently recorded in the CA1 than in the CA3 area and were not seen in the subiculum or dentate area. These experiments indicate that a glutamatergic-independent, synchronous GABA-mediated potential which is elicited by 4AP in the adult rat hippocampus continues to occur in the presence of TEA. In addition, concomitant application of these K(+)-channel blockers induces a novel type of prolonged field-potential discharge as well as spreading depression-like episodes. Since all synchronous potentials (including spreading depression-like episodes) were abolished by bicuculline methiodide, we conclude that their occurrence is presumably dependent upon the post-synaptic activation of GABAA receptors located on neuronal and glial elements. As excitatory synaptic transmission was nominally blocked under our experimental conditions, we also propose that rises in [K+]o and consequent redistribution processes are per se sufficient to make all types of synchronous activity propagate.

4-Aminopyridine↗
PubMed1996

Extracellular free potassium and calcium during synchronous activity induced by 4-aminopyridine in the juvenile rat hippocampus.

1. Field potential recordings and measurements of the extracellular concentration of free K+ ([K+]o) and Ca2+ ([Ca2+]o) were made during application of 4-aminopyridine (4-AP, 50 microM) in hippocampal slices that were obtained from 11- to 32-day-old rats. 2. Spontaneous field potentials recorded under this experimental condition in the CA3 stratum radiatum of slices from rats < 23 days old consisted of interictal (duration, 0.2-1.4 s; intervals of occurrence, 0.9-3.4 s) and ictal epileptiform discharges (duration, 5-46 s; intervals of occurrence, 22-259 s) and negative-going potentials that often preceded the onset of ictal discharge. Ictal activity became rare in slices from rats > 25 days old. 3. The negative-going potential (which also corresponded to the ictal discharge onset) was associated with [K+]o increases to 9.4 +/- 3.6 mM (mean +/- S.D.) from 3.25 mM baseline (n = 11 slices). [K+]o remained elevated at 5-6 mM throughout the ictal event. Decreases in [Ca2+]o (from 1.8 mM baseline to 1.3 +/- 0.1 mM, n = 7) were observed during the ictal discharge. 4. Interictal and ictal discharges were abolished by the non-N-methyl-D-aspartate (NMDA) receptor antagonist 6-cyano-7-nitroquinoxaline-2, 3-dione (CNQX, 10 microM). CNQX and the NMDA receptor antagonist 3-((+/-)-2-carboxypiperazine-4-yl)propyl-1-phosphonic acid (CPP) did not influence negative-going potentials or the associated [K+]o increases (peak values were 8.7 +/- 3.2 mM, n = 8), that were blocked, however, by bicuculline methiodide (BMI, 10 microM). 5. The mu-opioid receptor agonist (D-Ala2,N-Me-Phe4,Gly5-ol)-enkephalin (DAGO, 10 microM) which inhibits GABA release from interneurons, prevented the occurrence of both GABA-mediated synchronous potentials and subsequent ictal discharges (n = 6) as well as the [K+]o elevations. DAGO effects were antagonized by naloxone (10 microM; n = 4). 6. The GABA-mediated [K+]o elevations changed as a function of age. In hippocampal slices obtained from 11- to 17-day-old rats, peak values of 10.6 +/- 2.0 mM (n = 10) and half-width durations of 8.7 +/- 1.3 s (n = 7) were observed. In slices obtained from 25- to 32-day-old animals these parameters were 5.2 +/- 0.5 mM (n = 13) and 4.6 +/- 1.1 s (n = 4), respectively. 7. This study shows that, in the juvenile rat hippocampus, 4-AP induces a glutamatergic independent synchronous potential that is due to GABA released from inhibitory terminals and is associated with an increase in [K+]o. This [K+]o elevation undergoes age-dependent changes, and is instrumental in synchronizing neurons thus initiating prolonged epileptiform discharges.

4-Aminopyridine↗
PubMed1996

Synchronous GABA-mediated potentials and epileptiform discharges in the rat limbic system in vitro.

Application of 4-aminopyridine (4AP, 50 microM) to combined slices of adult rat hippocampus-entorhinal cortex-induced ictal and interictal epileptiform discharges, as well as slow field potentials that were abolished by the mu-opioid agonist [D-Ala2,N-Me-Phe4,Gly-ol5] enkephalin (DAGO, 10 microM) or the GABAA receptor antagonist bicuculline methiodide (BMI, 10 microM); hence, they represented synchronous GABA-mediated potentials. Ictal discharges originated in the entorhinal cortex and propagated to the hippocampus, whereas interictal activity of CA3 origin was usually recorded in the hippocampus. The GABA-mediated potentials had no fixed site of origin or modality of propagation; they closely preceded (0.2-5 sec) and thus appeared to initiate ictal discharges. Only ictal discharges were blocked by the antagonist of the NMDA receptor 3,3-(2-carboxypiperazine-4-yl)propyl-1-phosphonate (CPP, 10 microM), whereas the non-NMDA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 microM) abolished all epileptiform activities. The GABA-mediated potentials continued to occur synchronously in all regions even after concomitant application of CNQX and CPP. [K+]o elevations were recorded in the entorhinal cortex during the ictal discharge (peak values = 13.9 +/- 0.9 mM) and the synchronous GABA-mediated potentials (peak values = 4.2 +/- 0.1 mM); the latter increases were presumably attributable to postsynaptic GABAa-receptor activation because they were abolished by DAGO or BMI. Their role in initiating ictal activity was demonstrated by using DAGO, which abolished both GABA-mediated synchronous potentials and ictal discharges. These data indicate that NMDA-mediated ictal discharges induced by 4AP originate in the entorhinal cortex; such a conclusion is in line with clinical evidence obtained in temporal lobe epilepsy patients. 4AP also induces GABA-mediated potentials that spread within the limbic system when excitatory transmission is blocked and may play a role in initiating ictal discharge by increasing [K+]o.

4-Aminopyridine↗
PubMed1996

Developmental features of 4-aminopyridine induced epileptogenesis.

4-Aminopyridine (4-AP, 50 microM), perfused in rat hippocampal slices from postnatal days 2-30 (P2-P30), induced in the CA3 area the appearance of spontaneous epileptiform discharges, short (interictal-like) and sustained (ictal-like), as well as slow potential. The duration of epileptiform discharges decreased and their rate of occurrence (frequency) increased with maturation: their duration during the 1st postnatal week was 4-6 times longer and their frequency 5 times lower, compared to those of the 4th postnatal week. Ictal discharges gradually disappeared at the end of the 4th postnatal week. Spontaneous synchronous activity-as a whole-often appeared in clusters separated by equal or longer length inactive periods, during the first two postnatal weeks. At the same period, ictal discharges were often followed by repetitive afterdischarges, forming sequences which lasted 0.7-1.5 min. Sectioning experiments showed that epileptiform discharges were generated in CA3, and their presence in CA1 depended on the integrity of CA1-CA3 synaptic connections. In conclusion, these findings demonstrate that (i) immature CA3 can generate synchronous epileptiform discharges as early as P2, (ii) such discharges are longer lasting and more complex during the early developmental stages and (iii) there are two time points (end of 2nd, end of 4th postnatal weeks), when maturational changes alter the epileptogenic properties of immature hippocampus.

4-Aminopyridine↗
PubMed1996

Reverberation of chloride-dependent synaptic potentials in the rat entorhinal cortex in vitro.

The spontaneous activity generated by rat entorhinal neurons during application of 4-aminopyridine (4AP; 50 microM) was studied with intracellular and extracellular field-potential recordings in an vitro slice preparation. Long-lasting depolarizations (LLDs) with amplitudes of 15 +/- 7.6 mV (mean +/- SD; n = 14) and durations of 1.65 +/- 0.77 s (n = 14) occurred at 0.036 +/- 0.01/s (n = 14). Each LLD was followed by a rhythmic sequence of depolarizing potentials (up to 22 events) with amplitudes of 4-30 mV, durations of 40-500 ms and frequency of 0.9 +/- 0.2/s (n = 14). These intracellular potentials were mirrored by negative-going field potentials, suggesting that they represented synchronous events. Membrane input resistance decreased by 79-86% during both LLDs and subsequent rhythmic depolarizations. Intracellular injection of steady depolarizing or hyperpolarizing current modified the amplitude of these potentials in a similar manner: the reversal potential of the LLDs and of the rhythmic depolarizations was -66.4 +/- 4 mV and -67.9 +/- 3.2 mV, respectively (n = 7). Intracellular injection of Cl- increased the amplitude of both types of potentials. Spontaneous LLDs continued to occur during application of the non-N-methyl-D-aspartate (NMDA) receptor antagonist 6-cyano-7-nitro-quinoxaline-2,3-dione (10 microM), a procedure that abolished the subsequent rhythmic depolarizations (n = 3). LLDs were blocked by further addition of the gamma-aminobutyric acid (GABA)A receptor antagonist bicuculline methiodide (10 microM, n = 3). Our findings demonstrate that during 4AP application entorhinal neurons generate glutamatergic-independent LLDs as well as synchronous, Cl(-)-dependent depolarizations that reverberate through non-NMDA-mediated excitatory circuits.

4-Aminopyridine↗
PubMed1996

Extracellular K+ accumulations and synchronous GABA-mediated potentials evoked by 4-aminopyridine in the adult rat hippocampus.

Transient changes in extracellular potassium concentration ([K+]o) and field potentials were evoked by 4-aminopyridine (4-AP; 50-100 microM) and recorded with ion-selective microelectrodes in CA1b, CA3b and dentate sectors of adult rat hippocampal slices. Long-lasting field potentials recurred at a frequency of approximately 1/60 s (0.016 +/- 0.003 Hz) in association with increases in [K+]o which were largest and most sustained in the dendritic regions where afferent fibers terminate (dentate > CA1 > CA3) and in the hilus. In stratum radiatum of CA1 or stratum moleculare of the dentate these fields had a peak amplitude of 1.4 +/- 0.29 mV, duration 8.3 +/- 1.6 s, and were accompanied by increases in [K+]o of 1.8 +/- 0.22 mM that lasted 32 +/- 5.5 s (n = 17 slices). Interictal epilentiform potentials, which were brief (< 0.2 s) and more frequent at approximately 1/3 s (0.30 +/- 0.02Hz) were also present in CA1, CA3 and the hilus and associated with small increases in [K+]o (< or = 0.5 mM, duration < or = 2 s). Interictal activity was blocked by 6-cyano-7-nitroquinoxalone-2,3-dione (CNQX; 5-20 microM); the slow, less frequent potentials were resistant to both CNQX and DL-2-amino-5-phosphonovaleric acid (APV; 50 microM) and reversibly blocked (or attenuated by approximately 80%) by bicuculline methiodide (BMI) (25-100 microM). The BMI-sensitive potentials were also abolished by baclofen (100 microM), an effect which was reversed by 2-OH-saclofen (100 microM). Focal application of KCI or GABA in the absence of 4-AP evoked long-lasting field and [K+]o potentials which were similar to those evoked by 4-AP but more sustained. The proportional relationship between the amplitudes of field and K+ potentials with GABA closely resembled that observed for 4-AP; in contrast the slope of KC1-evoked responses was lower. Our results demonstrate that in the adult rat hippocampus 4-AP induces in many different regions accumulations of [K+]o in synchrony with the long-lasting field potentials, which are known to correspond to an intracellular long-lasting depolarization of the pyramidal cells. These changes are smaller than those which occur in the immature rat hippocampus--which may be related to differences in Na-K-ATPase and susceptibility to seizures. These events involve the activation of GABAA receptors, are under the modulatory control of GABAB receptors, and likely arise from the activity of GABAergic interneuron and/or afferent terminals. The long-lasting field potentials appear to reflect mainly the direct depolarizing actions of GABA and to much more limited extent the associated accumulation of [K+]o.

4-Aminopyridine↗
PubMed1996

Functional and pharmacological properties of human neocortical neurons maintained in vitro.

The availability of neocortical tissue obtained during brain surgery has allowed for detailed studies of the membrane and synaptic properties of neurons maintained in vitro in a slice preparation. Many of the findings obtained in these studies are summarized here. The majority of the basic electrophysiological properties appear to be similar when human and rodent neurons are compared. However, some notable exceptions regarding specific membrane properties have been reported. Since the majority of the material used in these studies is obtained from epileptic patients, several neuroscientists have tried to determine whether this tissue retains any sign of epileptogenicity when analyzed in vitro. Abnormal synaptic activity was only seen in a fraction of neurons near identified anatomical foci, including tumors, or within neocortical areas that displayed abnormal electrographic activity in situ. This cellular activity included both the presence of all-or-none and graded synaptic bursts. Epileptiform activity comparable to that seen in rodent tissue has been obtained in vitro using several pharmacological procedures including the disinhibition and the Mg(2+)-free model. In conclusion, electrophysiological and pharmacological studies of the human neocortex obtained during surgery have so far been unsuccessful in isolating any definite cellular mechanism that may account for the expression of the epileptiform activity in situ. Nevertheless, these studies have provided valuable information on the cellular and synaptic properties of human neocortex under normal conditions, and following experimental procedures capable of increasing neuronal excitability.

Cells, Cultured↗
PubMed1996

Epileptiform activity induced by pilocarpine in the rat hippocampal-entorhinal slice preparation.

An in vitro slice preparation of combined hippocampus and entorhinal cortex from adult rats was used to study the modalities of generation and propagation, as well as the pharmacological properties of the epileptiform activity induced by the muscarinic agonist pilocarpine (10 microM). Simultaneous field potentials recordings were made from the medial entorhinal cortex and from the dentate gyrus, CA3 and CA1 subfields. Pilocarpine application induced two types of interictal epileptiform discharges. The first occurred in the entorhinal cortex and consisted of bursts of population spikes lasting 408 +/- 135 ms (n = 20 slices) and repeating at a rate of 0.26 +/- 0.07 Hz (n = 20); this interictal activity propagated to the hippocampus via the perforant path. The second type was only observed in CA3 and CA1 subfields, had shorter duration (82 +/- 16 ms; n = 20) and occurred at a higher rate (1.42 +/- 0.7 Hz; n = 20) than the first type. Ictal epileptiform discharges (duration: 11.5 +/- 4.1 s; rate: 0.002 +/- 0.0009 Hz; n = 10) were also seen in the entorhinal cortex, from where they propagated to the dentate, CA3 and CA1 via the hippocampal trisynaptic loop as revealed by latency analysis and lesion experiments. Ictal and interictal discharges of entorhinal origin disappeared in the hippocampal sectors, but continued to occur in the entorhinal cortex following a cut of the perforant path (n = 5). Fast interictal discharges observed solely in the hippocampus originated in CA3, since sectioning the Schaffer collaterals made them disappear in CA1 (n = 7). All types of epileptiform activity disappeared during application of the non-N-methyl-D-aspartate receptor antagonist 6-cyano-7-nitroquinoxaline-2, 3-dione (10 microM; n = 7). By contrast, the N-methyl-D-aspartate receptor antagonist 3-3(2-carboxy-piperazine-4-yl) propyl-1-phosphonate (10 microM) abolished ictal discharges in the entorhinal cortex and reduced the duration of the interictal events recorded in this area (n = 7). Interictal discharges originating from CA3 continued to occur at a higher rate than in control during application of this N-methyl-D-aspartate receptor antagonist. Our study confirms that the combined hippocampal-entorhinal slice preparation represents a suitable model for understanding the modalities of origin and propagation of epileptiform activity within the limbic system. In this in vitro preparation, the entorhinal cortex is the site of origin for ictal discharges. Moreover, the different types of epileptiform activity induced by this muscarinic agonist have specific, structure-dependent pharmacological profiles. These results are discussed in relation to those obtained in vivo.

Animals↗