Effects of arginine vasopressin on water space in astrocytes and in whole brain.
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Animals↗
Biomedical subjects
L Hertz
Publications and source records attributed to L Hertz.
Noradrenaline effects on pyruvate decarboxylation: correlation with calcium signaling.
Noradrenaline effects on the rate of metabolism of pyruvate to acetyl coenzyme A, catalyzed by the pyruvate dehydrogenase complex, was measured in primary cultures of mouse astrocytes as rate of production of labeled CO(2) from 1-[(14) C]pyruvate in the absence of competing glucose in the medium. The subtype specificity of a noradrenaline-stimulated increase in rate of CO(2) formation was identical to that for noradrenaline-induced increase in free intracellular calcium ([Ca(2+)](i)), suggesting a causal relationship between these two phenomena. The noradrenaline-induced stimulation of pyruvate decarboxylation was abolished in the presence of 10 mM magnesium chloride in the medium, combined with the omission of calcium, a procedure known to prevent an increased [Ca(2+)] in the cytosol from raising intramitochondrial [Ca(2+)]. Thus, the stimulation of metabolic flux through the reaction catalyzed by the pyruvate dehydrogenase complex appears to result from an increase in intramitochondrial [Ca(2+)] ions in astrocytes. Such a mechanism for stimulation of the same enzyme has been convincingly demonstrated in other cell types, primarily heart muscle and hepatic cells, but it has not previously been demonstrated in any cell type from the central nervous system.
Adrenergic Agonists↗
Astrocytes: glutamate producers for neurons.
In order for the brain to use the common amino acid glutamate as a neurotransmitter, it has been necessary to introduce a series of innovations that greatly restrict the availability of glutamate, so that it cannot degrade the signal-to-noise ratio of glutamatergic neurons. The most far-reaching innovations have been: i) to exclude the brain from access to glutamate in the systemic circulation by the blood-brain barrier, thereby making the brain autonomous in the production and disposal of glutamate; ii) to surround glutamatergic synapses with glial cells and endow these cells with much more powerful glutamate uptake carriers than the neurons themselves, so that most released transmitter glutamate is rapidly inactivated by uptake in glial cells; iii) to restrict to glial cells a key enzyme (glutamine synthetase) that is involved in the return of accumulated glutamate to neurons by amidation to glutamine, which has no transmitter activity, and can be safely released to the extracellular space, returned to neurons and deaminated to glutamate; iv) to restrict to glial cells two key enzymes (pyruvate carboxylase and cytosolic malic enzyme) that are involved in, respectively, de novo synthesis (from glucose) of the carbon skeleton of glutamate, and the return of the carbon skeleton of excess glutamate to the metabolic pathway for glucose oxidation. As a consequence of these innovations, neurons constantly require new carbon skeletons from glial to sustain their TCA cycle. When these supplies are withdrawn, neurons are unable to generate amino acid transmitters and their rate of oxidative metabolism is impaired. Given the commensalism that exists between neurons and glia, it may be fruitful to view brain function not just as a series of interactions between neurons, but also as a series of interactions between neurons and their collaborating glial cells.
Animals↗
Does the 'mystery of the extra glucose' during CNS activation reflect glutamate synthesis?
The hypothesis is proposed that the repeatedly demonstrated rise in local cerebral blood flow and glucose utilisation during neuronal activation, without a corresponding increase in oxygen utilisation, may reflect glutamine formation from glucose, followed by complete oxidative degradation of glutamate along complex and extended, but well described pathways, known to operate in the brain. The former process requires large amounts of glucose but little oxygen. The latter utilises oxygen but no additional glucose, is a prolonged process and so at any one time involves only a small increase in the rate of oxygen utilisation which is difficult to demonstrate experimentally.
Brain↗
Hexokinase in astrocytes: kinetic and regulatory properties.
Hexokinase (HK, EC 2.7.1.1) is a key enzyme in the control of brain glucose metabolism. The regulatory role of HK in different neural cell types has not been elucidated. In this study we determined some kinetic and regulatory properties of HK in mouse cerebrocortical astrocytes in primary culture. Astroglial HK showed an absolute requirement for Mg-ATP and D-glucose. The pH optimum of HK was between 7.4 and 8.0. For astroglial HK, the Km for Mg-ATP was approximately 208 microM and Vmax approximately 35.4 mU/mg protein. At levels higher than 0.2 mM, D-glucose-1,6-bisphosphate, a known regulator of glycolysis, inhibited astroglial HK in a concentration-dependent manner, with an IC50 of approximately 0.4 mM; at 1.2 mM, it almost completely inhibited HK activity. The results obtained for astroglial HK are compatible with those reported for the highly purified preparations of brain HK. These data are of direct relevance to the assessment of glycolytic flux and its regulation in astrocytes.
Animals↗
Chronic treatment with lithium and pretreatment with excess inositol reduce inositol pool size in astrocytes by different mechanisms.
Chronic treatment with a lithium salt is the classical treatment for manic-depressive disorder. It is hypothesized that the therapeutic action of lithium is caused by its inhibition of inositol phosphatases which leads to a relative deficiency of inositol and, therefore, an impairment of inositol recycling and production of precursor for the second messengers inositol triphosphate (IP3) and diacylglycerol (DAG). However, peculiarly enough, treatment with high doses of inositol also has an antidepressant effect. In the present work, we have studied the acute and chronic effects of lithium and of excess inositol, in separation or together, on accumulation of 50 microM [3H]inositol (a physiologically relevant concentration) into primary cultures of mouse astrocytes. Two parameters were investigated: (1) rate of unidirectional uptake across the cell membrane (measured during short-term exposure to the radioisotope), and (2) magnitude of the intracellular pool of inositol, equilibrating with extracellular inositol (measured during long-term exposure to the radioisotope). Inositol uptake was highly concentrative and occurred with a Km of approximately 500 microM and a Vmax of 1.5 nmol/min/mg protein. The uptake rate was not affected by either acute or chronic treatment with LiCl (or both), but it was substantially reduced ('down-regulated') after pretreatment with a high concentration of inositol. The inositol pool size was decreased to a similar extent as the uptake rate by previous exposure to excess inositol. In spite of the fact that inositol uptake rate was unaffected by lithium, the magnitude of the inositol pool was significantly decreased by chronic treatment with a pharmacologically relevant concentration of LiCl (1 mM), but not by treatment with lower concentrations. This decrease is likely to reflect a reduction in either inositol synthesis or replenishment of inositol from IP3, due to the inhibition of inositol phosphatases by the lithium ion. In agreement with the different mechanisms by which lithium and pretreatment with excess inositol appear to reduce the pool size of inositol, the effects of pretreatment with excess inositol and of LiCl were additive. It is noteworthy that both effects could be observed in astrocytes, suggesting that there might be a significant astrocytic target during clinical treatment.
Animals↗
Peroxide-scavenging deficit underlies oligodendrocyte susceptibility to oxidative stress.
Previous work showed that the susceptibility of oligodendroglial progenitors to oxidative stress is related to their low reduced-glutathione (GSH) and high iron contents. This suggests that these cells have a poor ability to scavenge peroxides. All peroxides are scavenged by glutathione peroxidase. Glutathione peroxidase activity requires GSH as an electron donor resulting in the formation of oxidized-glutathione. Cellular GSH content is dependent upon synthesis as well as reduction of oxidized-glutathione. The objectives of the present study were to compare several parameters important in the ability to scavenge peroxides between astrocytes and oligodendroglia. Three stages of oligodendroglial differentiation were examined: the proliferative oligodendrocyte progenitor, the proliferative oligodendroblast, and the post-mitotic oligodendrocyte. We demonstrate that oligodendroglia at all stages of differentiation have less than one-half the content of GSH compared to astrocytes. This low level of GSH is due in part to a lower rate of GSH synthesis in oligodendroglia compared to astrocytes and in part to their having only one-half of the glutathione reductase activity of astrocytes. Glutathione peroxidase activity of oligodendroglia is less than 15% of that found in astrocytes. The low GSH and concomitant low glutathione peroxidase activity would tend to maintain peroxides at levels that are dangerously high if iron is released from iron stores. Oligodendroglia have high iron stores, and thus these findings emphasize how vulnerable the oligodendroglial lineage is to perturbations that result in oxidative stress.
Animals↗
Functional studies in cultured astrocytes.
Studies using primary cultures of astrocytes have made essential contributions to the understanding of astrocytic functions and neuronal-astrocytic interactions. The purposes of this article are to (i) outline principles and methodologies used in the preparation of such cultures and caveats for the interpretation of the observations made; (ii) summarize astrocytic functions in turnover of the amino acid transmitters glutamate and gamma-aminobutyric acid (GABA), in energy metabolism and in Na+,K+-ATPase-catalyzed processes and emphasize the degree to which the observations have been confirmed in intact tissue; (iii) describe regulations of astrocytic functions by transmitters and by calcium channel activity; and (iv) indicate suggestions for future functional studies using astrocytes in primary cultures and emphasize that some of the conclusions about neuronal-astrocytic interactions reached on the basis of studies in cultured cells and confirmed in intact tissue may not yet have been completely integrated into general neuroscience knowledge.
Adenosine Triphosphatases↗
Mitochondrial heterogeneity in the brain at the cellular level.
Classically, compartmentation of glutamate metabolism in the brain is associated with the fact that neurons and glia exhibit distinct differences with regard to metabolism of this amino acid. The recent use of 13C-labeled compounds to study this metabolism in conjunction with the availability of cell type-specific tissue culture modes has led to the notion that such compartmentation may even be present in individual cell types, neurons as well as glia. To better understand and explain this, it is proposed that mitochondrial heterogeneity may exist resulting in tricarboxylic acid cycles with different properties regarding cycling rates and ratio as well as coupling to amino acid biosynthesis, primarily involving glutamate and aspartate. These hypotheses are evaluated in the light of current knowledge about mitochondrial structure and function.
Animals↗
Can experimental conditions explain the discrepancy over glutamate stimulation of aerobic glycolysis?
Uncertainty reigns over whether or not glutamate uptake in astrocytes leads to strong stimulation of glucose utilization, measured as accumulation of radioactive deoxyglucose-6-phosphate. This is an important issue, not only because glutamate is the major excitatory transmitter, but also because it has been postulated that glutamate-induced stimulation of glycolysis links brain excitation with activation of energy production. The effect of glutamate on deoxyglucose utilization in cultured rat and mouse astrocytes grown in different media and incubated under various conditions during the deoxyglucose assay has, therefore, been studied. Under most conditions, no stimulation occurred but rather a decrease in deoxyglucose utilization during exposure to glutamate; under certain conditions, the contribution of non-metabolized deoxyglucose to the intracellular 14C signal was significant.
Aerobiosis↗
Effects of a low-voltage static electric field on energy metabolism in astrocytes.
Mouse astrocytes (glial cells) in primary cultures were exposed to a low-voltage static DC electric field with no current flow and thus with no generation of magnetic fields. The electric field altered the rate of glycolysis, measured by 2-deoxyglucose accumulation. The magnitude and direction of this effect depended on the polarization of the field and the applied voltage. The maximum effect was an increase of approximately 30%, which occurred with field across the cells at an intensity that can be calculated to be 0.3 mV/cm or less. Reversal of the polarization converted the stimulation to a small but statistically significant inhibition.
Animals↗
Complex roles of glutamate in the Gibbs-Ng model of one-trial aversive learning in the new-born chick.
Glutamate is the most widespread excitatory transmitter in the CNS and is probably involved in LTP, a neural phenomenon which may be associated with learning and memory formation. Intracerebral injection of large amounts of glutamate between 5 min and 2.5 min after passive avoidance learning in young chicks inhibits short-term memory, which occurs between 0 and 10 min post-learning in a three-stage model of memory formation first established by Gibbs and Ng(25) [Physiol. Behav. 23:369-375; 1979]. This effect may be attributed to non-specific excitation. Blockade of glutamate uptake by L-aspartic and beta-hydroxamate also abolishes this stage of memory, provided the drug is administered within 2.5 min of learning. Interference with either production of percursors for transmitter glutamate in astrocytes or with glutamate receptors is also detrimental to memory formation, but the effects appear much later. After its release from glutamatergic neurons, glutamate is, to a large extent, accumulated into astrocytes where it is converted to glutamine, which can be returned to glutamatergic neurons and reutilized for synthesis of transmitter glutamate, and partly oxidized as a metabolic substrate. The latter process leads to a net loss of transmitter glutamate which can be compensated for by de novo synthesis of a glutamate precursor alpha-ketoglutarate (alpha KG) in astrocytes, a process which is inhibited by the astrocyte-specific toxin fluoroacetate (R. A. Swanson, personal communication). Intracerebral injection of this toxin abolishes memory during an intermediate stage of memory processing occurring between 20 and 30 min post-training (50) [Cog. Brain Res, 2:93-102; 1994]. Injection of methionine sulfoximine (MSO), a specific inhibitor of glutamine synthetase, which interferes with the re-supply of transmitter glutamate to neurons by inhibition of glutamine synthesis in astrocytes, has a similar effect. This effect of MSO is prevented by intracerebral injection of glutamate, glutamine, or a combination and alpha KG and alanine. MSO must be administered before learning, but does not interfere with acquisition since short-term memory remains intact. Administration of either the NMDA antagonist AP5, the AMPA antagonist DNQX, or the metabotropic receptor antagonist MCPF, also induces amnesia. Memory loss in each case does not occur until after 70 min post-training, during a protein synthesis-dependent long-term memory stage which begins at 60 min following learning. However, to be effective, AP5 must be administered within 60 s following learning, MCPG before 15 min post-learning, and DNQX between 15 and 25 min after learning. Together, these findings suggest that learning results in an immediate release of glutamate, followed by a secondary release of this transmitter at later stages of processing of the memory trace, and that one or both of these increases in extracellular glutamate concentration are essential for the consolidation of long-term memory. Since both fluoroacetate and MSO act exclusively on glial cells, the findings also show that neuronal-glial interactions are necessary during the establishment of memory.
Animals↗
Cell culture studies of the role of elevated extracellular glutamate and K+ in neuronal cell death during and after anoxia/ischemia.
During vascular insults in the brain (stroke) the extracellular concentrations of glutamate and K+ increase. It is well acknowledged that the increase in glutamate contributes to the death of neuronal cells at an earlier time than they would have succumbed to energy deprivation as such, but the origin of the released glutamate is not known and cannot easily be studied in the brain in vivo. We have therefore resorted to cell culture studies which have shown that the neuronal rate of formation of glutamate from glutamine is substantially increased during anoxia, especially in glutamatergic neurons. This increase is further enhanced in the presence of excess K+. Phenylsuccinate, a compound that decreases formation of glutamate from glutamine in glutamatergic neurons, counteracts the increase in glutamate formation and, by doing so, improves cell survival. Astrocytes (glial cells) in neuronal-astrocytic co-cultures to some extent protect against anoxic neuronal damage by accumulating glutamate and thus keeping the extracellular glutamate concentration lower than in isolated neuronal cultures.
Animals↗
Inhibition of noradrenaline stimulated increase in [Ca2+]i in cultured astrocytes by chronic treatment with a therapeutically relevant lithium concentration.
Chronic treatment of mouse astrocytes in primary cultures with 1 mM lithium chloride for 7-14 days decreased the basal level of free cytosolic calcium concentration ([Ca2+]i) from 50-70 nM to approximately 70% of this value and reduced the increase in [Ca2+]i caused by exposure to 1 microM noradrenaline (normally to 500-700 nM) by almost one half. A similar, but much smaller, response to serotonin was unaffected by chronic treatment with lithium. Acute exposure to lithium (30 min) had no effect on either basal or noradrenaline stimulated [Ca2+]i. The dependence on chronic, versus acute treatment suggests that this effect may be related to the therapeutic effect of lithium as a mood-stabilizing drug, which likewise requires chronic treatment. Since good evidence is found that noradrenaline increases [Ca2+]i by activation of the phosphoinositol second messenger system the present findings are also consistent with literature data that lithium acts by interfering with this system.
Animals↗
Glutathione release and catabolism during energy substrate restriction in astrocytes.
This study examined the effect of simulated ischemia (deprivation of both oxygen and substrate) on astrocyte reduced-glutathione (GSH). We have demonstrated that under normoxic conditions there is no GSH efflux from living astrocytes; this suggests that the high levels of GSH in astrocytes in vivo are not available for neighbouring neural cells. Under simulated ischemia there is release of GSH from astrocytes only when astrocytes die. Furthermore, when astrocytic energy stores are depleted GSH is catabolized, such that after 12 h of simulated ischemia approximately 20% of GSH is catabolized. This GSH catabolism can be increased at an earlier time by causing increased ATP utilization through activating the sodium pump either by introducing glutamate into the culture medium or by raising medium potassium. Since GSH is catabolized into glycine, glutamate and cysteine, the latter two amino acids being neurotoxic, our findings indicate that the high levels of GSH in astrocytes may be used by these cells to survive ischemic insults, but the catabolism of GSH may result in increased neuronal damage.
Adenosine Triphosphate↗
Astrocyte-neuron interaction during one-trial aversive learning in the neonate chick.
During two specific stages of the Gibbs-Ng model of one-trial aversive learning in the neonate chick, we have recently found unequivocal evidence for a crucial involvement of astrocytes. This evidence is metabolic (utilization of the astrocyte-specific energy store, glycogen, during normal learning and inhibition of memory formation by the astrocyte specific metabolic inhibitors, fluoroacetate and methionine sulfoximine) as well as physiological (abolition of memory formation in the presence of ethacrynic acid, an astrocyte-specific inhibitor of cellular reaccumulation of potassium ions). These findings are discussed in the present review in the framework of a more comprehensive description of metabolic and physiological neuronal-astrocytic interactions across an interstitial (extracellular) space bounded by minute processes from either cell type.
Animals↗
Acute and chronic effects of potassium and noradrenaline on Na+, K+-ATPase activity in cultured mouse neurons and astrocytes.
Acute and chronic effects of elevated extracellular concentrations of potassium ions ([K+]0) and/or noradrenaline were studied in homogenates of primary cultures of mouse astrocytes, from the cerebral cortex or the spinal cord, and of primary cultures of mouse cerebral cortical neurons. NA+, K+-ATPase activity in cerebral cortical astrocytes showed a Km value of 1.9 mM with confidence limits of 1.3-2.9 mM and a Vmax of 5.4 mumol/h/mg protein with confidence limits of 3.3-8.1 mumol/h/mg protein. Due to the high Km value, the activity of the enzyme was significantly increased by an increase in [K+]0 in the interval 5-12 mM. In cerebral cortical neurons, Vmax was lower (1.77 +/- 0.06 mumol/h/mg protein) but the affinity was higher (Km 0.43 +/- 0.8 mM). With these kinetics, there is no stimulation of enzyme activity when [K+]0 is increased beyond control levels. In spinal cord astrocytes, the relative effect of increasing [K+]0 above 6 mM was larger than in cerebral astrocytes but the absolute activity of the enzyme was lower. Na+, K+-ATPase activity in both types of astrocyte was stimulated by noradrenaline and its beta-adrenergic subtype agonist isoproterenol but mainly or exclusively at 6 mM [K+]0. Noradrenaline also caused a stimulation in cortical neurons, but at non-physiological K+ concentrations this stimulation was converted to an inhibition, and isoproterenol had no stimulatory effect. Chronic exposure of cerebral cortical astrocytes to elevated [K+]0 caused a decrease in Na+, K+-ATPase activity when enzyme activity in the cells was subsequently measured at normal [K+]0. During exposure to 30 mM [K+]0 this "down-regulation" took place within 10 min. Conversely, chronic exposure to reduced [K+]0 led to an increase in Na+, K+-ATPase activity. Chronic exposure to noradrenaline had no significant effect but there was a tendency towards an increase.
Adrenergic alpha-Agonists↗
Inhibition of glutamine synthetase activity prevents memory consolidation.
Methionine sulfoximine, a specific inhibitor of the exclusively glial enzyme glutamine synthetase, was shown, at a concentration of 3.5-4.5 mM, to prevent consolidation of memory for a passive avoidance task in day-old chicks. Provided the drug was administered 5-20 min before the learning task, significant retention loss was observed from the normal time of onset of the second of three postulated stages in the memory formation sequence but the drug had to be administered considerably earlier. The amnestic effect of methionine sulfoximine was successfully counteracted by L-glutamine (10 mM) and monosodium glutamate (4 mM), and also by a cocktail of alpha-ketoglutarate (5 mM) and alanine (5 mM). This effect of methionine sulfoximine is attributed to its blockade of the production of glutamine via the glutamate-glutamine cycle, leading to a reduced capacity of neurons to replenish their transmitter glutamate.
Alanine↗