Sándor Borbély

Lateral entorhinal cortex lesions rearrange afferents, glutamate receptors, increase seizure latency and suppress seizure-induced c-fos expression in the hippocampus of adult rat.

The entorhinal cortex (EC) provides the predominant excitatory drive to the hippocampal CA1 and subicular neurones in chronic epilepsy. Here we analysed the effects of one‐sided lateral EC (LEC) and temporoammonic (alvear) path lesion on the development and properties of 4‐aminopyridine‐induced seizures. Electroencephalography (EEG) analysis of freely moving rats identified that the lesion increased the latency of the hippocampal seizure significantly and decreased the number of brief convulsions. Seizure‐induced neuronal c‐fos expression was reduced in every hippocampal area following LEC lesion. Immunocytochemical analysis 40 days after the ablation of the LEC identified sprouting of cholinergic and calretinin‐containing axons into the dentate molecular layer. Region and subunit specific changes in the expression of ionotropic glutamate receptors (iGluRs) were identified. Although the total amount of AMPA receptor subunits remained unchanged, GluR1flop displayed a significant decrease in the CA1 region. An increase in NR1 and NR2B N‐methyl‐d‐aspartate (NMDA) receptor subunits and KA‐2 kainate receptor subunit was identified in the deafferented layers of the hippocampus. These results further emphasize the importance of the lateral entorhinal area in the spread and regulation of hippocampal seizures and highlight the potential role of the rewiring of afferents and rearrangement of iGluRs in the dentate gyrus in hippocampal convulsive activity.

Modification of ionotropic glutamate receptor-mediated processes in the rat hippocampus following repeated, brief seizures.

The seizure-induced molecular and functional alterations of glutamatergic transmission in the hippocampus have been investigated. Daily repeated epileptic seizures were induced for 12 days by intraperitoneal administration of 4-aminopyridine (4-AP; 4.5 mg/kg) in adult Wistar rats. The seizure symptoms were evaluated on the Racines scale. One day after the last injection, the brains were removed for in vitro electrophysiological experiments and immunohistochemical analysis. The glutamate receptor subunits NR1, NR2A, NR2B, GluR1, GluR1(flop), GluR2, and KA-2 were studied using the histoblotting method. The semi-quantitative analysis of subunit immunoreactivities in hippocampal layers was performed with densitometry. In the hippocampus, increase of GluR1, GluR1(flop) and NR2B immunostaining was observed in most of the areas and layers. The significant decrease of GluR2 staining intensity was observed in the CA1 and dentate gyrus. Calcium permeability of hippocampal neurons was tested by a cobalt uptake assay in hippocampal slices. The uptake of cobalt increased in the CA1 area and dentate gyrus, but not in the CA3 region following 4-AP treatment. Effects of AMPA and NMDA (N-methyl-d-aspartate) glutamate receptor antagonists (1-(4-aminophenyl)-4-methyl-7,8-methylenedioxy-5H-2,3-benzodiazepine hydrochloride (GYKI 52466) and D-APV respectively) were measured in hippocampal slices using extracellular recording. Analysis of the population spikes revealed the reduced effectiveness of the AMPA receptor antagonist GYKI 52466, while the effect of the NMDA receptor antagonist d-(2R)-amino-5-phosphonovaleric acid was similar to controls. The results demonstrated that repeated convulsions induced structural and functional changes in AMPA receptor-mediated transmission, while NMDA and kainate receptor systems displayed only alterations in receptor subunit composition.

Repeated 4-aminopyridine induced seizures diminish the efficacy of glutamatergic transmission in the neocortex

Systemic administration of the potassium channel blocker 4-aminopyridine (4-AP) elicits acute convulsions. Synchronized tonic-clonic activity develops during the first hour after the treatment. However, subsequent chronic spontaneous seizures do not appear which suggests changes in neuronal excitability. The aim of our present work was to evaluate alterations in the glutamatergic transmission in the somatosensory cortex of rats following daily, brief convulsions elicited by 4-AP treatment. Changes in general neuronal excitability and pharmacological sensitivity of glutamate receptors were tested in ex vivo electrophysiological experiments on brain slices. In parallel studies quantitative changes in subunit composition of glutamate receptors were determined with immunohistoblot technique, together with the analysis of kainate induced Co2+ uptake. The results of our coordinated electrophysiological, receptor-pharmacological and histoblot studies demonstrated that repeated, daily, short convulsions resulted in a significant decrease of the general excitability of the somatosensory cortex together with changes in ionotropic glutamate receptor subunits. The relative inhibitory effect of the AMPA receptor antagonist, however, did not change. The NMDA receptor antagonist exerted somewhat stronger effect in the slices from convulsing animals. 4-AP pretreatment resulted in the attenuation of kainate induced Co2+ uptake, which suggests either reduction in non-NMDA receptors numbers or reduction in their Ca2+ permeability. Repeated seizures decreased GluR1-4 AMPA receptor subunit levels in all cortical layers with a relaitve increase in GluR1 subunits. While the principle NR1 NMDA receptor subunit showed no significant change, the staining density of NR2A subunit increased. These changes in ionotropic glutamate receptors are consistent with reduced excitability at glutamatergic synapses following repeated 4-AP induced seizures.

Laminar analysis of initiation and spread of epileptiform discharges in three in vitro models

Overexcitation of neuronal networks in some forebrain structures and pathological synchronization of neuronal activity play crucial role in epileptic seizures. Seizure activity can be elicited experimentally by different convulsants. Because of various distribution of excitatory and inhibitory connections in the neocortex there might be laminar differences in seizure sensitivity. Current source density (CSD) analysis or immunocytochemical c-Fos localization offer suitable tools to localize increased activation of neurons during seizure. In the present experiments, interictal epileptiform activity elicited by 4-aminopiridine, bicuculline or Mg(2+)-free solution was recorded with a 16-channel multielectrode assembly in different layers of the somatosensory cortex, and CSDs were calculated. Parallel c-Fos immunocytochemistry was applied. Each convulsant elicited characteristic activation pattern. 4-aminopiridine induced relatively short discharges, which were associated with a huge sink in layer V, the sink and source pattern was relatively simple. Mg(2+)-free solution elicited the longest discharges, sinks appeared typically in the supragranular layers II and III than quickly distributed toward layers V and VI. Bicuculline induced rather similar seizure pattern as Mg(2+)-free solution, but the amplitudes of field potentials were larger, while the durations shorter. The peak of c-Fos activation, however, was not parallel with the largest electrical activation. Larger amount of stained cells appeared in layers II and III in 4-aminopiridine and bicuculline, respectively. In Mg(2+)-free solution the highest c-Fos activity was detected in upper layer VI. Long-lasting cellular effects do not always correspond to the largest electrical responses, which are primarily determined by the activation of asymmetrical pyramidal neurons. Interneurons, which possess more symmetric process arborisation, play less important role in the generation of field potentials, although they may be intensively activated during seizure.

Protein kinase D promotes plasticity-induced F-actin stabilization in dendritic spines and regulates memory formation

PKD regulates the stabilization of the F-actin network within dendritic spines upon chemically induced plasticity changes and is needed for proper hippocampal LTP and spatial memory formation.

Synthetic calpain activator boosts neuronal excitability without extra Ca2

Earlier we have shown that an equimolar mixture of calpastatin subdomains A and C (19 amino acids each) strongly activates m-calpain in vitro. In the present work we developed a membrane-permeable activator system, by conjugating an oligo-arginine tail to both peptides. We tested calpain activation as well as synaptic excitability on rat brain slices ex vivo. In hippocampal slices both basic excitability and long-term synaptic efficacy were significantly increased upon treatment with the activator. We propose that the activator peptide conjugates can be used with any mammalian cell, to specifically challenge the calpain system apparently without raising cytoplasmic Ca2+. Such an effector may be a useful tool in dissecting intracellular mechanisms involving the calpain system.

In vitro intrinsic optical imaging can be used for source determination in cortical slices.

In the last decades intrinsic optical imaging has become a widely used technique for monitoring activity in vivo and in vitro. It is assumed that in brain slices the source of intrinsic optical signals (IOSs) is the change in light scattering caused by cell swelling or shrinkage. The aim of the present study was to find a correlation between electrical activity and parallel optical characteristics, elicited by 4‐aminopyridine‐containing or Mg2+‐free medium in rat cortical brain slices. Electrophysiological signals and reflected light alterations were recorded during spontaneous seizure activity. Current source density (CSD) analysis was performed on the electrophysiological records. Direct correlation analysis of IOS to CSD was made, and source distribution provided by IOS and CSD methods was compared by determining Matthews correlation coefficient. The gradual development of seizure‐like activity elicited the reduction of light reflectance. The main findings of our experiments are that long‐term epileptiform activity resulted in persistent alteration in IOSs of brain slices. The observed IOS pattern remained stable after 1 h incubation in convulsants. The pattern of IOS shows good correlation with the data obtained from the CSD analysis. Persistent IOS changes provide information about the area‐specific changes of basic excitability, which can serve as a background for ictal and interictal‐like epileptiform activity. We can conclude that changes in IOSs correlate well with electrophysiological recordings under different conditions. Our experiments provide evidence that underlying synchronised neuronal processes produce parallel alterations in IOSs and electrophysiological activity.

Toward the Full Quantum Dynamical Description of Photon Induced Processes in D2

The dissociative ionization (multiphoton regime) of the D2+ ion by ultrashort laser pulses has been studied theoretically using ab initio calculations. The combined ionization and dissociation spectrum was explored for fixed molecular axis orientations. In accordance with previous investigations, the dominant features in the obtained joint energy spectrum were multiphoton peaks. In addition to this, in the present work, photoelectron angular distributions were analyzed as well. By performing a partial wave analysis for each multiphoton peak, we have identified the number of absorbed photons. Moreover, we also found that the angular distribution can significantly change inside a multiphoton peak as a function of electron and nuclear kinetic energy.

An efficient numerical discretization method for the study of the H2+ in intense laser fields

We present an accurate and efficient numerical discretization approach for the calculation of the electronic eigenstates of the H2+ molecule, and the transition dipole moments between them. The obtained high precision wave functions and dipole moments are planned to be used during the direct numerical solution of the time-dependent Schrodinger equation (TDSE) for the H2+ molecule irradiated with ultrashort laser pulses.

Over‐the‐barrier ionization of H2O by intense ultrashort laser pulses

The ionization of the H2O molecule in intense ultrashort electric fields was studied theoretically. Ionization probability densities were calculated using classical and quantum mechanical approaches. The water molecule is treated within the hydrogenic approximation. Classical calculations were carried out using the classical trajectory Monte Carlo method, while quantum mechanical calculations are based on the sudden strong field approximation. A good agreement between classical and quantum calculations was found.