Perspectives on spreading depression

Spreading depression (SD) consists of a transient suppression of all neuronal activity that spreads slowly across regions of gray matter. The paper is divided into three parts. Martins-Ferreira describes 30 years of research on SD in the isolated retina. Much of this work has relied on the prominent intrinsic optical signals that accompany SD in the retina. By inducing SD to propagate in circles with a velocity of 3.7 mm min⁻¹, it is possible to investigate the finely balanced electrochemical equilibrium that maintains the traveling wave. SD is accompanied by a slow negative extracellular voltage and ion movements that are greatest in the inner plexiform layer of the retina. Nedergaard discusses the role of astrocytes in SD propagation. Astrocytes mediate slowly moving waves of intracellular Ca²⁺ increase, for which gap junctions are essential. SD is accompanied by entry of Ca²⁺ into cells and fails when gap junctions are blocked. SD, however, is blocked by glutamate receptor antagonists but glial Ca²⁺ waves are not. Astrocytic Ca²⁺ waves are probably involved in the initiation of SD but other factors, including K⁺, glutamate and purinergic receptors, are necessary for sustained propagation. Nicholson describes studies on the different preparations that helped clarify the role of extracellular space in SD. It has long been known that extracellular K⁺ reaches levels of 50 mM or more during SD. Studies with ion-selective microelectrodes showed that extracellular Na⁺ and Cl⁻ fall by as much as 100 mM during SD, and water leaves the extracellular space. Further work showed that extracellular Ca²⁺ falls 10-fold during SD and significant changes in extracellular pH and ascorbate occur. These studies imply that large perturbations of the extracellular milieu occur during SD and are an essential part of the interlocking cascade of events that produce this still mysterious phenomenon.     

Cortical spreading depression recorded from the human brain using a multiparametric monitoring system

The number of parameters (i.e., EEG or ICP-intracranial pressure) routinely monitored under clinical situations is limited. The brain function analyzer described in this paper enables simultaneous, continuous on-line monitoring of cerebral blood flow (CBF) and volume (CBV), intramitochondrial NADH redox state, extracellular K⁺ concentrations, DC potential, electrocorticography and ICP from the cerebral cortex. Brain function of 14 patients with severe head injury (GCS ≤ 8), who were hospitalized in the neurosurgical or general intensive care unit was monitored using this analyzer. Leao cortical spreading depression (SD) has been reported in many experimental animals but not in the human cerebral cortex. In one of the patients monitored, spreading depression was observed. This is the first time that spontaneous repetitive cortical SD cycles have been recorded from the cerebral cortex of a patient suffering from severe head injury. Typical SD cycles appeared 4–5 h after the beginning of monitoring this patient. During the first 3–4 cycles the responses of this patient were very similar to the responses to SD recorded in normoxic experimental animals. Electrocorticography was depressed whereas extracellular K⁺ levels increased. The metabolic response to spreading depression was characterized by oxidation of intramitochondrial NADH concomitant to a large increase in CBF. During brain death, an ischemic depolarization, characterized by decrease in CBF and an irreversible increase in extracellular K⁺, was recorded.     

Phenytoin and retinal spreading depression

Several studies indicate that spreading depression is fundamentally related to seizure marches and to the aura of classical migraine. Moreover, recent investigations call attention to its possible relevance in clinical disturbances associated with brain ischemia, trauma, and hypoglycemia. The anticonvulsant phenytoin has been shown to protect the nervous tissue from the effects of anoxia and ischemia. These properties suggest that phenytoin should be able to counteract spreading depression. Therefore, we investigated its effect on spreading depression elicited by mechanical or chemical (KCl) stimulation, in isolated chick retinas. The results showed that phenytoin: (1) increases the threshold concentration of KCl to initiate the phenomenon; (2) decreases the velocity of propagation of spreading depression; (3) shortens considerably the duration of the slow potential, ionic (K+, Ca2+, Cl-), and volume changes of the extracellular compartment during spreading depression. Possible mechanisms underlying the observed effects are discussed.     

Energy failure in astrocytes increases the vulnerability of neurons to spreading depression

A neuroprotective role of astrocytes has been hypothesized, but the mechanism is debated and in vivo evidence is limited. To test this hypothesis, a sublethal stressor (spreading depression) and fluorocitrate (FC), a selective inhibitor of the astrocytic Krebs cycle, were used in urethane-anaesthetized adult rats. Neuronal damage was assessed 24 h after treatment with silver stain and immunoreactivity for a 72-kDa heat-shock protein. ATP levels and mitochondrial aconitase activity, a marker indicating exposure to reactive oxygen species, were measured after 4 and 24 h. Spreading depression alone did not affect ATP levels, mitochondrial aconitase activity, or induce neuronal injury in the cortex. Local or intraventricular injection of FC significantly decreased ATP levels and mitochondrial aconitase activity, but did not produce neuronal damage. In animals receiving injections of FC and then spreading depression, there was evidence of significant neuronal stress and damage. Isocitrate, which bypasses the metabolic inhibition produced by FC, prevented all of the changes seen after the combination of FC and spreading depression. One-hour pretreatment with dimethyl sulfoxide (a scavenger of hydroxyl radicals), deferoxamine (an iron chelator) or fructose-1,6-bisphosphate also blocked inactivation of mitochondrial aconitase, ATP depletion and the neuronal damage induced by FC and spreading depression. These experiments demonstrate that inhibition of the metabolism of astrocytes, with a decrease in ATP levels, will increase the susceptibility of neurons to the stress induced by spreading depression. The neuroprotective effects of dimethyl sulfoxide, deferoxamine and fructose-1,6-bisphosphate suggest that oxidative stress contributes to the neurotoxicity in this situation.     

Gabapentin suppresses cortical spreading depression susceptibility

Cortical spreading depression (CSD) is an intense depolarization wave implicated in the pathophysiology of brain injury states and migraine aura. As Ca_v2.1 channels modulate CSD susceptibility, we tested gabapentin, which inhibits Ca_v2.1 through high-affinity binding to its α₂δ subunit, on CSD susceptibility in anesthetized rats. Gabapentin, 100 or 200 mg/kg, elevated the electrical threshold for CSD and diminished recurrent CSDs evoked by topical KCl, when administered 1 hour before testing. With its favorable safety and tolerability profile, gabapentin may have a role in suppression of injury depolarizations in stroke, intracranial hemorrhage, and traumatic brain injury.     

Regenerative glutamate release by presynaptic NMDA receptors contributes to spreading depression

Spreading depression (SD) is a slowly propagating neuronal depolarization that underlies certain neurologic conditions. The wave-like pattern of its propagation suggests that SD arises from an unusual form of neuronal communication. We used enzyme-based glutamate electrodes to show that during SD induced by transiently raising extracellular K⁺ concentrations ([K⁺]_o) in rat brain slices, there was a rapid increase in the extracellular glutamate concentration that required vesicular exocytosis but unlike fast synaptic transmission, still occurred when voltage-gated sodium and calcium channels (VGSC and VGCC) were blocked. Instead, presynaptic N-methyl-D-aspartate (NMDA) receptors (NMDARs) were activated during SD and could generate substantial glutamate release to support regenerative glutamate release and propagating waves when VGSCs and VGCCs were blocked. In calcium-free solutions, high [K⁺]_o still triggered SD-like waves and glutamate efflux. Under such a condition, glutamate release was blocked by mitochondrial Na⁺/Ca²⁺ exchanger inhibitors that likely blocked calcium release from mitochondria secondary to NMDA-induced Na⁺ influx. Therefore presynaptic NMDA receptor activation is sufficient for triggering vesicular glutamate release during SD via both calcium entry and release from mitochondria by mitochondrial Na⁺/Ca²⁺ exchanger. Our observations suggest that presynaptic NMDARs contribute to a cycle of glutamate-induced glutamate release that mediate high [K⁺]_o-triggered SD.     

Factors affecting the oxygen balance in the awake cerebral cortex exposed to spreading depression

Oxygen balance was evaluated in the cerebral cortex, using the surface fluorometry technique of the intramitochondrial NADH redox state, exposed to various physiological and pathological situations. Using flexible fiber optic light guide, connected to the brain surface via a cemented holder, the measurements were done continuously from the awake rat and gerbil. In few experiments the NADH redox state was correlated with the electrical and ionic activity (measured by surface K⁺ and DC electrodes). Three different animal models were used in the study: the adult rat, the very young rat (20 g) and the adult gerbil. Those 3 models were used in studying the effect of hypoxia, partial ischemia, and anesthesia on the metabolic and ionic activities measured from the awake brain. Spreading cortical depression (elicited by topical KCl solution) was used as a standard stimulation of the ionic and metabolic activities of the cerebral cortex. Two typical metabolic responses to spreading depression (SD) were recorded, namely ‘oxidation cycle’ and ‘reduction cycle’ depending upon the ability of the tissue to compensate for the extra amount of oxygen needed for the higher mitochondrial activity.It was found that the adult rat brain showed oxidation cycles in most conditions (besides partial ischemia), while the young rat and the gerbil brains were much more sensitive to the various perturbations of the brain and exhibited reduction cycle (as a response to SD) under all pathological situations tested.We conclude from our detailed studies that the type of response to SD, as measured by NADH surface fluorometry, represents the oxygen balance which exists in the tissue under various conditions.     

Cortical spreading depression transiently activates MAP kinases

Cortical spreading depression (CSD) has been shown to have neuroprotective effects when administered in advance of cerebral ischemia. The mechanism by which CSD induces its neuroprotective effect however remains to be elucidated. Since MAP kinases have been shown to impart neuroprotection in ischemic preconditioning paradigms, we attempted to determine the role CSD may have in the activation of MAPK. We show that CSD is capable of increasing the phosphorylation of ERK in a MEK-dependent manner. This phosphorylation is, however, transient, as phosphorylated ERK levels return to control levels 45 min after 2 h of CSD elicitation. Immunohistochemical analysis reveals that the phosphorylated form of ERK is located ubiquitously in cells of the CSD-treated cortex while CSD-elicited MEK phosphorylation resides solely in the nuclei. These data suggest that CSD may act via the MAP kinase pathways to mediate preconditioning.     

Leao's spreading depression: synaptic regulation of a diffuse self-oscillating process in the central nervous system

The evidence reviewed in the paper shows that the spreading depression (SD) belongs to the mass of closely packed neurons which can support qualitatively new phenomena occurring at a more primitive level of cellular interaction. The latter becomes prepotent under extreme conditions, overloading the local energy supply and homeostatic mechanisms. The two levels of neural integration, the highly heterogeneous specifically connected synaptic level and the statistically uniform diffusively organized nonsynaptic level, are always present in all forms of brain activity and mutually influence each other. While phenomena mediated by nonsynaptic interaction overrule the synaptic ones under resting conditions, excessive synaptic excitation may either switch the affected brain region from synaptic to nonsynaptic interaction (spike-triggered SD) or assert the synaptic interaction over the nonsynaptic one (functional blockade of SD). Nonsynaptic interaction is an inherent property of brain, an obligatory consequence of the trade-off between maximal packing density and independent function of individual elements of the system which sets definite limits to the maximum activation of synaptic processes, contributes to slow synchronization of neural populations and plays an important role in various instances of brain pathology. The phenomena generated at this level belong to the family of autowaves, share their typical properties and can be described by similar formal rules.     

Responses of rat brain to induced spreading depression following exposure to carbon monoxide

Until recently carbon monoxide (CO) was known only for its noxious effects. Exposure to CO results in an autoregulatory increase in cerebral blood flow (CBF). Little information is available on brain energy metabolism under low CO concentrations and on the effect of CO on the stimulated brain. In this study cortical spreading depression (SD) was induced in order to cause transient brain depolarization and increased energy demand. The multisite assembly (MSA), which contains four bundles of optical fibers for monitoring the intramitochondrial NADH redox state and tissue reflectance as well as four DC electrodes enabling measurement from four consecutive points on the cerebral cortex, was used to measure energy metabolism and the propagation of SD waves during exposure to CO. CBF in the contralateral hemisphere was measured using the laser Doppler technique. Three experimental groups of animals were examined: SD was induced during exposure to 1000 ppm CO, immediately after exposure to CO and 90 min after cessation of exposure to CO. Three control groups were also examined, in which the animals underwent the same procedures but were not exposed to CO. In all animals exposure to CO was followed by a significant increase in CBF. The greatest effect was found when SD was induced immediately after cessation of exposure to CO. SD wave frequency decreased when induced immediately after exposure to CO, whereas it increased when SD was induced 90 min after exposure. The amplitude of the NADH oxidation waves and their integral were smaller during SD induced immediately after exposure to CO. The DC potential did not change, suggesting that CO did not affect the SD initiation mechanism but rather resulted in energy depletion during recovery from SD. This study demonstrates that even at a concentration of 1000 ppm CO interferes with the metabolic activity of the brain during repolarization of the SD-induced negativity.