The ability of neuroglia to buffer local increases of extracellular K(+) has been known from in vitro studies. This property may confer on these cells an active role in the modulation and spreading of cortical oscillatory activities. We addressed the question of the spatial buffering in vivo by performing single and double intraglial recordings, together with measures of the extracellular K(+) and Ca(2+) concentrations ([K(+)](out) and [Ca(2+)](out)) in the cerebral cortex of cats under ketamine and xylazine anesthesia during patterns of slow sleep oscillations and spike-wave seizures. In addition, we estimated the fluctuations of intraglial K(+) concentrations ([K(+)](in)). Measurements obtained during the slow oscillation indicated that glial cells phasically take up part of the extracellular K(+) extruded by neurons during the depolarizing phase of the slow oscillation. During this condition, the redistribution of K(+) appeared to be local. Large steady increases of [K(+)](out) and phasic potassium accumulations were measured during spike-wave seizures. In this condition, [K(+)](in) rose before [K(+)](out) if the glial cells were located at some distance from the epileptic focus, suggesting faster K(+) diffusion through the interglial syncytium. The simultaneously recorded [Ca(2+)](out) dropped steadily during the seizures to levels incompatible with efficient synaptic transmission, but also displayed periodic oscillations, in phase with the intraseizure spike-wave complexes. In view of this fact, and considering the capability of K(+) to modulate neuronal excitability both at the presynaptic and postsynaptic levels, we suggest that the K(+) long-range spatial buffering operated by glia is a parallel synchronizing and/or spreading mechanism during paroxysmal oscillations.

The mechanisms that control the periodicity of spontaneous epileptiform cortical potentials were investigated in the in vitro isolated guinea pig brain preparation. A brief intracortical application of bicuculline in the piriform cortex induced spontaneous interictal spikes (sISs) that recurred with high periodicity (8.5 Ϯ 3.1 sec, mean Ϯ SD). Intracellular recordings from principal neurons showed that the early phase of the inter-sIS period is caused by a GABAb receptor-mediated inhibitory potential. The late component of the interspike period correlated to a slowly decaying depolarization abolished at membrane potentials positive to Ϫ32.1 Ϯ 5.3 mV and was not associated with membrane conductance changes. Specific pharmacological tests excluded the contribution of synaptic and intrinsic conductances to the late inter-sIS interval. Recordings with ion-sensitive electrodes demonstrated that sISs determined both a rapid increase in extracellular K ϩ concentration (0.5-1 mM) and an extracellular alkalinization (0.05-0.08 pH units) that slowly decayed during the inter-sIS period and returned to control values just before a subsequent sIS was generated. These observations were not congruous with the presence of a silent period, because both extracellular increase in K ϩ and alkalinization are commonly associated with an increase in neuronal excitability. Extracellular alkalinization could be correlated to an sIS-induced intracellular acidification, a phenomenon that reduces cell coupling by impairing gap junction function. When intracellular acidification was transiently prevented by arterial perfusion with NH 4 Cl (10-20 mM), spontaneous ictal-like epileptiform discharges were induced. In addition, the gap junction blockers octanol (0.2-2 mM) and 18-␣-glycyrrethinic acid (20 M) applied either via the arterial system or locally in the cortex completely and reversibly abolished the sIS. The results reported here suggest that the massive cell discharge associated with an sIS induce a strong inhibition, possibly secondary to a pH-dependent uncoupling of gap junctions, that regulates sIS periodicity.

There is a growing interest for connections between the central nervous system (CNS) and systemic immune and/or inflammatory responses. The inflammatory cytokine IL-1 has a wide spectrum of targets, which include the CNS. Intracerebroventricular as well as systemic administration of IL-1 were reported to induce central effects including fever (1), slow-wave sleep (2), anorexia (3), and activation of the hypothalamus-pituitary axis leading to release of adrenal corticosteroids (4, 5). Moreover, different brain cells such as astrocytes, microglia, and neurons can respond and/or have been found to contain various cytokines like IL-1, IL-2, IL-6, and TNF, and there is evidence that these factors act on neuronal survival, growth, and differentiation (1, 3, 6-10).It is also possible that some of the systemic activities of IL-1 are, at least in part, centrally mediated . In fact, early reports indicated that central administration of crude leukocytic endogenous mediators (presumably containing, among other cytokines, IL-1) induced an increase of acute-phase proteins (11) . The mechanism by which centrally administered IL-1 can activate the synthesis of hepatic acute-phase proteins is still unknown, and the present study was aimed at revisiting this early observation by investigating how intracerebroventricularly administered rIL-1 induced a systemic response. Our attention was focused on IL-6, since this cytokine, induced by IL-1, plays a crucial role in the acute-phase response as a hepatocyte-stimulating factor, and the levels of circulating IL-6 were reported to correlate with the levels of acute-phase proteins in some infective and inflammatory diseases (12)(13)(14).

Interleukin-1 (IL-1) and IL-1 receptors are constitutively expressed in normal brain. IL-1 increases non-rapid eye movements (NREM) sleep in several animal species, an effect mediated in part by interactions with the serotonergic system. The site(s) in brain at which interactions between IL-1 and the serotonergic system increase NREM sleep remain to be identified. The dorsal raphe (DRN) is the origin of the major ascending serotonergic pathways to the forebrain, and it contains IL-1 receptors. This study examined the hypothesis that IL-1 increases NREM sleep by acting at the level of the DRN. IL-1beta (0.25 and 0.5 ng) was microinjected into the DRN of freely behaving rats and subsequent effects on sleep-wake activity were determined. IL-1beta 0.5 ng increased NREM sleep during the first 2 h post-injection from 33.5 +/- 3.7% after vehicle microinjection to 42.9 +/- 3.0% of recording time. To determine the effects of IL-1beta on electrophysiological properties of DRN serotonergic neurons, intracellular recordings were performed in a guinea-pig brain stem slice preparation. In 26 of 32 physiologically and pharmacologically identified serotonergic neurons, IL-1beta superfusion (25 ng/mL) decreased spontaneous firing rates by 50%, from 1.6 +/- 0.2 Hz (before IL-1beta superfusion) to 0.8 +/- 0.2 Hz. This effect was reversible upon washout. These results show that IL-1beta increases NREM sleep when administered directly into the DRN. Serotonin enhances wakefulness and these novel data also suggest that IL-1beta-induced enhancement of NREM sleep could be due in part to the inhibition of DRN serotonergic neurons.

We have recently demonstrated that the myelinolytic lesions in the spinal cord (SC) of rats made deficient in vitamin B(12) (cobalamin) (Cbl) through total gastrectomy (TG) are tumor necrosis factor-alpha (TNF-alpha)-mediated. We investigate whether or not permanent Cbl deficiency, induced in the rat either through TG or by chronic feeding of a Cbl-deficient diet, might modify the levels of three physiological neurotrophic factors-epidermal growth factor (EGF), vasoactive intestinal peptide (VIP), and somatostatin (SS)-in the cerebrospinal fluid (CSF) of these rats. We also investigated the ability of the central nervous system (CNS) in these Cbl-deficient rats to synthesize EGF mRNA and of the SC to take up labeled Cbl in vivo. Cbl-deficient rats, however the vitamin deficiency is induced, show a selective decrease in EGF CSF levels and an absence of EGF mRNA in neurons and glia in various CNS areas. In contrast, radiolabeled Cbl is almost exclusively taken up by the SC white matter, but to a much higher degree in totally gastrectomized (TGX) rats. Chronic administration of Cbl to TGX rats restores to normal both the EGF CSF level and EGF mRNA expression in the various CNS areas examined. This in vivo study presents the first evidence that the neurotrophic action of Cbl in the CNS of TGX rats is mediated by stimulation of the EGF synthesis in the CNS itself. It thus appears that Cbl inversely regulates the expression of EGF and TNF-alpha genes in the CNS of TGX rats.