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The supportive role of glial cells on neuronal function and survival has been studied in anesthetized rats by using the selective gliotoxin fluorocitrate. Disabling glia operation reproduced many features of ischemic penumbra. An initial mild acidosis and increased interstitial potassium but not glutamate was followed after 3-4 h by repetitive spreading depression waves. These gradually provoked higher levels of acidosis, potassium and glutamate, gradual neuronal function decay and finally, neuron death. We conclude that neurons become highly vulnerable to spreading depression waves only in absence of normal glia operation. Our findings directly associate early glial disfunction to neuronal loss and lead to new insights for the understanding of ischemic pathology.

The neuroprotective effect of acidic fibroblast growth factor (aFGF) has been analysed in a rat model of seizures-associated brain damage. We report that after treatment with a convulsivant dose of Kainic acid, systemically administered aFGF prevents neuronal degeneration in specific brain areas, mainly in the hippocampal formation. Our findings extend the potential pharmacological use of fibroblast growth factors and afford new data to understand the neurophysiology of these proteins.

Spreading depression of Leão (SD) can be provoked by numerous nonspecific mechanical, electrical, and chemical stimuli. A similar, if not identical, phenomenon can be provoked by hypoxia. SD is characterized by drastic depolarization of neurons, severe reduction of membrane resistance, and redistribution of ions across cell membranes. Glial cells also depolarize but retain membrane resistance. Tetraethylammonium hastens the onset of hypoxic SD but reduces the sustained potential shift and K+ outflow from cells; 4-aminopyridine also accelerates SD but has no effect on the voltage shift. N-Methyl-D-aspartate receptor antagonists delay the onset of SD, while nickel and cobalt reduce the amplitude of SD-related redistribution of Ca2+. Yet, no specific blocker of SD has been found. Microdialysis of high-K+ solution in hippocampal CA1 region induces recurrent waves of SD propagating semi-independently in adjacent tissue layers, and a prolonged unstable depressed state that has not previously been described. Neither the release of K+ ions nor of glutamate is the unique agent of SD propagation. On the basis of recent findings we propose a hypothetical sequence of events that reconcile many of the previously seemingly paradoxical observations.

The glutamate analogues N-methyl-D-aspartate (NMDA), kainic acid (KA), and quisqualic acid (QA), prepared in different hypertonic media, were perfused in vivo in the hippocampal CA1 field of rats using a microdialysis technique. Extracellular taurine levels, estimated after analysis of the taurine content of dialysates, increased during perfusion of all three agonists but varied according to the osmolarity of the medium. The NMDA-induced increase in extracellular taurine content was only slightly inhibited by perfusion of 150 and 300 mM sucrose. The KA-evoked increase was partially dependent on extracellular osmolarity, because addition of 50 and 150 mM sucrose caused a dose-dependent inhibition that was not augmented using higher sucrose concentrations. QA caused a taurine increase that was totally abolished by addition of 50 mM sucrose. These results indicate that the rise in extracellular taurine level elicited by QA and part of the increase elicited by KA are probably due to a release caused by the cellular swelling that these substances evoke, a finding substantiating the previously proposed osmoregulatory role of taurine. However, almost all the increase in extracellular taurine content caused by NMDA and all the osmotically insensitive part of the KA-evoked rise cannot be explained as release triggered by cell swelling and may reflect a function of taurine other than osmoregulation.

Taurine increases in brain extracellular space due to glutamate agonists were studied in vivo in the rat hippocampus using a dialysis technique, both in the absence and in the presence of glutamate receptor antagonists. Extracellular taurine levels increased during perfusions of agonists, listed in descending order of potency: kainate (KA), N-methyl-D-aspartate (NMDA), and quisqualate (QA). While taurine increases due to KA or QA perfusions were inhibited by 6,7-dinitro-quinoxaline-2,3-dione (DNQX), those induced by NMDA were abolished in the presence of 3-(carboxypiperazin-4-yl)-propyl-1-phosphonic acid (CPP). These results indicate that increases in extracellular taurine levels evoked by NMDA, KA or QA in the rat hippocampus are caused by activation of their specific receptors. Field potentials, concomitantly recorded, were quickly abolished during NMDA or KA perfusions (0.1 mM), while QA (0.25 mM) induced the appearance of bicuculline-like evoked responses. Since taurine has been proposed as an osmoregulatory substance in the rat brain, and cell swelling is known to be an early component of glutamate agonists neurotoxicity, the increases in extracellular taurine reported here could be due to taurine released through an osmoregulatory process, counteracting the neurotoxic cellular oedema induced by glutamate agonists.

6,7-Dinitro-quinoxaline-2,3-dione (DNQX, FG 9041), a new non-N-methyl-D-aspartate (NMDA) glutamate receptor antagonist, has been reported to block non-NMDA receptor-mediated excitatory amino acidic responses in cultured neurons. We have perfused this compound in vivo through a dialysis fiber placed in the CA1 regions of anesthetized rats to test its effects on CA1 field-evoked potentials. Perfusions of 25-100 microM DNQX completely abolished field excitatory postsynaptic potentials (EPSP) and orthodromic population spikes (PS). This effect was dose-dependent and was reversed after washing with fresh Krebs-Ringer-bicarbonate. Antidromic population spikes and fiber volley potentials were unaffected by perfusions of DNQX up to 100 microM. On the contrary, perfusion of 50 microM D-2-amino-5-phosphonovalerate, a specific NMDA receptor antagonist, left unchanged both field EPSP and orthodromic PS. Results demonstrate that low-frequency transmission at the Schaffer collaterals-CA1 synapse is mediated by non-NMDA glutamate receptors.

The effects of different K+ concentrations (3-100 mM) on both the extracellular amino acid levels and field potentials, evoked by perforant pathway stimulation, were studied 'in vivo' in the rat dentate gyrus by means of a brain dialysis device, formed by a hollow fiber plus a stainless-steel electrode. Perfusion with low K+ concentrations (3-12 mM; Krebs-Ringer bicarbonate) specifically enhanced the dialysate levels of taurine and concomitantly increased the population spike amplitude. High K+ concentrations in perfusate (greater than 25 mM) did not further increase the levels of taurine but enhanced both glutamate and gamma-aminobutyric acid levels, whereas the population spike diminished drastically. The absence of calcium ions in the perfusion liquid increased both basal and K+-enhanced taurine levels. The specific enhancement of extracellular taurine by physiological K+ concentrations may represent an autoregulative mechanism of nervous tissue excitability.

The effects of hypotonic media on extracellular free amino acid levels were studied 'in vivo' in the rat dentate gyrus by means of the brain dialysis technique. Extracellular taurine levels increased specifically during perfusions with Krebs-Ringer bicarbonate in which the NaCl concentration was reduced by 25 or 50 mmol/l (hypotonic solutions). These taurine increases were markedly reduced in the presence of furosemide. With further NaCl reductions the enhanced taurine levels remained stable, whereas other amino acids such as glutamate started in increase in a dose-dependent manner. Isoosmolar replacement of NaCl by sucrose did not affect extracellular amino acid levels. These results indicate the possible involvement of taurine in osmoregulatory processes in the brain.