In vitro and in vivo brain slice techniques were used to examine phencyclidine (PCP) effects on the lateral propagation of epileptiform field potentials (EFP) across adjacent areas of rat frontal neocortex. Epileptiform activity was induced by perfusing slices with Mg 2+-free artificial cerebrospinal fluid. Simultaneous field potential recordings of EFP were obtained from four microelectrodes placed 2-3 mm apart across coronal slices in the third layer. PCP, applied focally between recording sites, blocked rapid propagation across treated areas and resulted in the emergence of spatially separate, independent pacemakers. The characteristics of paroxysmal depolarization shifts did not change significantly by the blockade of lateral propagation of EFP. The same asynchronized pattern of EFP conduction was observed after local application of the N-methyl-D-aspartate (NMDA)-receptor antagonist DL-2-amino-5-phosphono-valeric acid. Local administration of haloperidol as well as NMDA before PCP application reversibly prevented appearance of multiple pacemakers. Focal application of dopamine produced an abnormal pattern of lateral conduction of EFP in 50 % of tested slices. Pacemaker failure as an indicator of functional impairment of cortical integration is the proposed mechanism for developing of schizophrenia-like psychosis associated with epilepsy. Abbreviations. APV: DL-2-amino-5-phosphono-valeric acid EEG:electroencephalogram EFP:epileptiform field potentials NMDA:N-methyl-D-aspartate PCP:phencyclidine SLPE:Schizophrenialike psychosis associated with epilepsy

Cortical spreading depression (CSD) occurrence has been suggested to be associated with seizures, migraine aura, head injury and brain ischemia-infarction. Only few studies identified CSD in human neocortical slices and no comprehensive study so far evaluated this phenomenon in human. Using the neocortical tissue excised for treatment of intractable epilepsy, we aimed to investigate CSD in human. CSD was induced by KCl injection and by modulating T-type Ca(2+) currents in incubated human neocortical tissues in an interphase mode. The DC-fluctuations were recorded by inserting microelectrodes into different cortical layers. Local injection of KCl triggered single CSD that propagated at 3.1+/-0.1 mm/min. Repetitive CSD also occurred spontaneously during long lasting application (5 h) of the T-type Ca(2+) channel blockers amiloride (50 microM) or NiCl(2) (10 microM) which was concomitant with a reversible extracellular potassium increase up to 50 mM. CSD could be blocked by the N-methyl-D-aspartate receptor antagonist 2-amino-5-phosphonovaleric acid in all cases. The results demonstrate that modulation of the Ca(2+) dynamics conditioned human neocortical slices and increased their susceptibility to generate CSD. Furthermore, these data indicate that glutamatergic pathway plays a role in CSD phenomenon in human.

Spreading depressions (SD) occur in association with ischaemia, epilepsy and migraine. Intracellular calcium oscillations have been suggested to be involved in the generation and propagation of SD. The present study was performed to study the mechanism of conditioning guinea pig hippocampal slices by the T-type calcium channel blockers NiCl2 and amiloride. SD-like fluctuations of DC potential were recorded by inserting microelectrodes into the CA1 and CA3 regions. The SD occurrence was significantly greater with 10 µmol/ l NiCl2 as well as with 25 and 50 µmol/ l amiloride than with other concentrations of these substances. The concentration response curve was inversely U-shaped with the maximum repetition rates of SDs being achieved at 10 µmol/ l NiCl2 as well as at 25 and 50 µmol/ l amiloride. SD occurrence could be completely blocked by the NMDA antagonist APV (10 µmol/ l) in all cases. These data demonstrate that modulation of the Ca2+ dynamics conditioned guinea pig hippocampal slices and increased their susceptibility to generate SD.

Generalized neurotransmitter overflow into the extracellular space, after cerebral ischemia, has been suggested to contribute to subsequent neuronal death. This study aims to investigate the striatal release of the neurotransmitters dopamine (DA), glutamate (Glu) and gamma-aminobutyric acid (GABA) by means of microdialysis, in a rat model for focal transient cerebral ischemia. Ischemia was induced by the application of 120 pmol endothelin-1 (Et-1), adjacent to the middle cerebral artery (MCA) in freely moving rats. Ischemia produced a large increase in extracellular striatal DA concentrations (2400%), Glu (5500%) and GABA (800%) concentrations. Laser Doppler flowmetry in anaesthetized rats, indicated that the blood flow within the striatum decreased by 75±11%. The period of sustained drop of blood flow, was dose-dependently related to the concentration Et-1 injected. Histological analysis of brain slices, taken from anaesthetized and conscious animals, indicated a 500 pmol dose of Et-1 was required to produce a similar infarct in anaesthetized rats to a 120 pmol dose of Et-1 in freely moving rats. The immediate drop in striatal blood flow, and the prompt increase of extracellular DA, after the micro-application of Et-1, were quite striking. This suggests that the DA release, rather than the Glu overflow may be the primary event initiating the cascade of processes ultimately leading to cell death and neurological deficits.

Taurine and glutamate were monitored by microdialysis technique during various cerebral insults: a. Application of K+ triggered a cortical spreading depression (CSD). Taurine and glutamate increased concomitantly but recovery of glutamate was faster than that of taurine. b. Application of NMDA induced also CSD but only taurine increased. c. Induction of an infarct triggered repetitive CSDs. Taurine increased rapidly whereas glutamate rose slowly starting with some delay. d. After induction of ischemia, taurine and glutamate increased after onset of depolarisation. The increase of glutamate occurred late after a small, transient increase in parallel with the depolarisation. These data suggest a close functional relationship between the changes of both amino acids. Therefore, they should be monitored together especially in clinical settings: during excitation, only taurine will increase; during overexcitation, taurine will also increase but to a higher maximum followed by a moderate rise of glutamate; after energy failure, taurine will accumulate to its highest level followed by a continuous rise of glutamate.

The K+-channel blocker tetraethylammonium (TEA) was applied to study its effects in self-sustaining, circling spreading depression in the chicken retina (RSD), Extracellular K+ (K-e(+)) activities and the direct-current (DC) signal were recorded using double-barrelled microelectrodes. Superfusion of TEA-concentrations of 10 to 250 mu M for 4 to 7 min reduced the RSD-associated DC amplitude (by 9 to 45%) and the maximum of the K-e(+) concentration (by 9 to 34%) in a dose-dependent manner, Propagation velocity of the RSD was lowered by 24%, At concentrations higher than 250 mu M TEA (0.5 to 10 mM), the propagation was slowed by more than 60%, after which the RSD disappeared, Recovery upon reperfusion with Ringer was immediate, These observations illustrate: 1) TEA affects the K-e(+) changes during RSD at very low concentrations. 2) The reduced K-e(+) transients are accompanied by a reduction of the DC shifts, 3) These changes of the electrical properties of the RSDs are paralleled by a reduction of the propagation velocity, 4) The effects of TEA are reversible, 5) The changes of these parameters occur dose-dependently, These data suggest a close relationship between the amplitudes of the ionic/electric changes during RSDs and the mechanisms of propagation. (C) 1998 Wiley-Liss, Inc.

The present study was performed in order to determine the oxygen consumption of a rat brain perfused with an artificial oxygen carrier. Measurements were performed prior to and 2 or 30 min after an ischemic period of 5 min. In addition, the energy-related metabolites were determined. Basal oxygen consumption and energy state were comparable to in vivo conditions. The tissue concentration of the energy metabolites decreased during ischemia and completely recovered during 30 min of reperfusion. The oxygen consumption was higher in the early phase of reperfusion than under pre-ischemic conditions. However, the oxygen consumption in the later phase of reperfusion was lower than the basal consumption. The data demonstrate that the addition of an artificial oxygen carrier to the perfusate provides sufficient amounts of oxygen to the in vitro preparation and that the measurement of the oxygen consumption during the post-ischemic reperfusion is a more sensitive parameter for the detection of emerging deficits than the measurement of the tissue levels of the energy metabolites.

The ability of retinal tissue to propagate "spreading depressions" (rSDs) depends on the endogenous properties of the tissue and requires an intact neuro-glial network. The phenomenon of rSDs, therefore, may be used as a tool to assess the functional integrity of brain tissue. We interfered with transmembranal Cl- movements in order to characterise the reactivity of the neuro-glial network subsequent to a pharmacological treatment. The rSDs were monitored by means of double-barrelled microelectrodes for de and extracellular Cl- recordings using the in vitro model of circling spreading depressions in the chicken retina. The application of the Cl-/HCO3-; exchange inhibitor DNDS caused the rSD related amplitude of the de signal as well as that of the Cl- signal to decrease until the point that rSDs finally ceased. The drug-induced changes of the de potential as well as those of the Cl- signal occurred in close correlation. This suggests that rSD related Cl- transients are an intrinsic property of de generating mechanisms. The data also show that modulation of only one of several possible mechanisms involved in the regulation of Cl- homeostasis as eg by DNDS is sufficient to prevent propagation of rSDs. Since propagation of rSDs requires the interaction of several excitatory and inhibitory processes, the present experiments indicate that the retinal tissue was made less susceptible to hyperexcitation by blocking the Cl-/HCO3-; exchanger with DNDS. Thus, Cl- exchange mechanisms seem to play a more important role in maintaining cellular excitability than taken into account so far. (C) 1997 Published by Elsevier Science Ltd.

Lubeluzole is a neuroprotective compound that has been shown to stereoselectively rescue sensorimotor function and reduce infarct size in a photochemical stroke model in rats. Tissue swelling, which occurs in the peri-infarct zone, is accompanied by a compensatory taurine release. Therefore, using a microdialysis technique, we aimed at measuring changes of extracellular concentrations of taurine in the peri-infarct zone and the effects of lubeluzole and its R-isomer. Lubeluzole blocked the increase of taurine in tissue immediately surrounding a photochemically induced thrombotic neocortical infarct. By contrast, the R-isomer was completely inactive. We hypothesize that lubeluzole may reduce osmoregulatory stress in peri-infarct tissue.

The measurement of cerebral extracellular lactate levels has been suggested to be used to monitor cerebral function in intensive care. However, although an increase of extracellular lactate levels is a sensitive parameter for increased cellular activity in general, it will be shown that its prognostic value is limited in regard to the severity of the impairment of cellular function. As an alternative, the measurement of the extracellular levels of inorganic phosphate (IP) or adenosine is proposed here: Whereas extracellular lactate levels increased rapidly to about the same extents during ischemia (IS) and spreading depression (SD), IP rose during IS only. Adenosine, on the other hand, increased during both events to a different degree. If, therefore, lactate was the only parameter to be monitored after a cerebral insult, the results would not allow to discriminate between a transient, spontaneously recovering event as a SD and a long-lasting or an irreversible loss of cell function as in persisting ischemia/hypoxia. The measurement of IP, therefore, seems to be more suitable than that of lactate or adenosine since IP will appear within the extracellular space only after a sustained failure of membrane function. Thus, the measurement of IP changes turned out to be the more useful parameter for intensive care supervision.

The chicken retina is an accessible piece of intact gray matter in which a self-sustained form of the 'Spreading Depression' (SD) wave can be easily elicited and recorded for many hours with double barrel ion-sensitive electrodes in the extracellular space. The blockade of glial (Müller) cell potassium channels with barium chloride added to the perfusing Ringer depressed both the negative potential shift typical of SDs and the velocity of spread. Moreover, there was separation of the extracellular increase of potassium and the drop in the extracellular potential: the peak of the potassium wave was increased, as well as its duration whereas the potential wave could be depressed to zero or even inverted to positive. By contrast the transient extracellular calcium drop could not be separated from the extracellular potential wave but appeared related to it: no transient calcium drop was observed when the negative potential was completely depressed or inverted. Both, the amplitude of the extracellular potential and extracellular calcium activity appeared to be important factors controlling the velocity of spread.

Long-term changes of learning behavior and of the striatal dopaminergic system were observed in a rat model of early postnatal hypoxia. Striatal dopamine (DA) concentration, K+-stimulated DA release from slices, and DA uptake into crude synaptosomal preparations (S1 fractions) were used as markers of the striatal DAergic system. Active avoidance learning was tested as behavioral criterion. Cyclodextrin and flunarizine were found to produce long-term effects on the DAergic system in control animals. While cyclodextrin normalized hypoxia-induced effects in DA release, flunarizine prevented those in DA uptake and improved avoidance learning.

pH sensitive microcelectrodes were used in combination with microdialysis (MD) technique to measure extracellular pH (pHe) and extracellular lactate (lace) within the cortex of rat brains during cortical spreading depression (SD). SD was induced by local K(+)-application and identified by DC recordings. It was accompanied by an extracellular acidification of 0.34 +/- 0.06 pH units and by a 2.8 +/- 0.80 fold increase of lace; the recovery of pHe took place within three phases, that of lace within 2 phases. The recovery of both parameters was complete about 45 min after the onset of SD. We conclude that the changes of lace and pHe are closely related. This indicates both lactate and protons to be transported in parallel.

Tissue levels of inorganic phosphate (iP-) and lactate (lac) increase during cerebral ischemia and cortical spreading depression (SD). Since cell membranes become leaky during these insults, iP- and lac were expected to leak into the extracellular space (ECS). In order to find out whether this occurs or does not, a microdialysis (MD) fiber was implanted into the cortex of anesthetized rats and extracellular lactate (lac(e)) and extracellular iP- (iPe-) were determined during various insults. Extracellular lactate increased to about the same extent during ischemia and SD. In contrast, iPe- increased during ischemia but not during SD. Instead, iPe- started to rise after SD and reached its maximum about 45 min later. The distinct pattern of iPe- in comparison to lac(e) during the above mentioned insults points to a qualitative difference of the underlying mechanisms: whereas lac appears within the ECS at any stressful situation, elevation of iP- within the ECS indicates depletion of energy stores in parallel to the lack of control of ion homeostasis.