Glucose Regulates Glutamate-Evoked Arachidonic Acid Release from Cultured Striatal Neurons

Abstract: l -Glutamate stimulates the liberation of arachidonic acid from mouse striatal neurons via the activation of N -methyl- d -aspartic acid (NMDA) receptors and by the joint stimulation of α-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) and metabotropic receptors. In this study, we investigated whether starving cultured mouse striatal neurons of glucose would modify glutamatergic receptor-mediated arachidonic acid release. Glucose deprivation for 30 min led to enhancement of the NMDA-evoked release of arachidonic acid, compared with that observed in the presence of glucose. This enhanced response depended on both the concentration of glucose and the length of time of glucose deprivation. The enhanced NMDA response appeared to result from both a release of glutamate and the subsequent additional release of arachidonic acid due to the activation of AMPA and metabotropic receptors. Indeed, the increased NMDA response was completely reversed when extracellular glutamate was enzymatically removed. Moreover, glucose deprivation potentiated the combined AMPA/metabotropic receptor-evoked release of arachidonic acid, even in the absence of extracellular glutamate. However, removing glucose did not improve the calcium rise induced by AMPA or NMDA. The ATP-evoked release of arachidonic acid from striatal astrocytes was not altered by glucose starvation. In summary, glucose deprivation affected two properties of striatal neurons: (a) it induced an NMDA-evoked release of glutamate from striatal neurons and (b) it selectively potentiated the AMPA/(1 S ,3 R )-1-aminocyclopentane-1,3-dicarboxylic acid-evoked release of [³H]arachidonic acid without altering the authentic NMDA-mediated response.     

Studies on cerebral energy metabolism during the course of galactose neurotoxicity in chicks

—At various times during a 2-day study, the levels of adenine nucleotides and selected glycolytic intermediates were determined in brains of chicks fed a diet containing d -galactose (40%, w/w). The levels of ATP and glucose 6-phosphate had decreased by 9 h after initiation of the diet, whereas those of fructose 1,6-diphosphate, 3-phosphoglycerate, l -α-glycerophosphate, and lactate were not reduced until after 18 h had elasped. Although glucose 1-phosphate was not appreciably affected, glucose and glycogen were depleted during the latter stages of the toxicity. The cerebral levels of 3′,5′-cyclic AMP and citrate did not differ significantly between the two dietary groups at 48 h. The changes in the levels of cerebral glycolytic intermediates and high-energy phosphates during ischemia indicated that the glycolytic rate was diminished in the chicks fed galactose and that high-energy phosphate compounds were depleted sooner than in controls. After intraperitoneal injection of [¹⁴C]glucose, the specific radioactivity and levels of glucose in the plasma from chicks fed either diet were similar, whereas they were significantly reduced in the brains from galactosefed animals. We suggest that galactose interferes with the uptake of glucose into the brain and that this mechanism may be an important factor in d -galactose-induced neurotoxicity in the chick.     

EFFECTS OF KAINIC ACID ON ION DISTRIBUTION AND ATP LEVELS OF STRIATAL SLICES INCUBATED IN VITRO

Abstract— To determine the mechanism of neurotoxicity of kainic acid, striatal slices (350μ) were incubated in oxygenated Krebs buffer with kainic acid and other depolarizing agents; and the alterations in the uptake and retention of ²²Na⁺, ⁸⁶Rb⁺ (as a measure of K ⁺), ³H_zO and the levels of ATP were determined. The excitatory amino acid, L-glutamate (10 mM) increases striatal slice uptake and retention of Na⁺, K⁺ and H₂O but decreases ATP levels whereas the neuroexcitant, A'-methyl aspartate, increases only Na⁺ and H₂O. Veratridine (100μM), which opens electrogenic sodium channels, and ouabain (100μM), which inhibits Na⁺-K⁺ ATPase, both elevate striatal Na⁺ and H₂O but considerably reduce K⁺ and ATP. The effects of these different depolarizing agents on the parameters examined are consistent with their mechanisms of actions and support the validity of this in vitro method. Although 10mM-kainate significantly depresses striatal K⁺ and ATP, lower concentrations of kainate (5mM-0.1μ) elevate striatal uptake of Na⁺ but do not markedly affect H₂O, K⁺ or ATP. Kainate (10mM-lμM) does not exhibit additivity with 10 mM-glutamate with respect to Na⁺ permeability but does significantly potentiate glutamate's ATP depleting effects. Injection of 10 nmol of kainate into the striatum in vivo causes a reduction in striatal ATP 1 h afterward which is comparable to that occurring in vitro with 10mM-kainate alone or with lower concentrations of kainate (≥1/μM) with 10 mM-glutamate. These results suggest that kainate alone is directly neurotoxic at 10mM or neurotoxic at lower concentrations in combination with the high intrasynaptic levels of glutamate on neurons receiving glutamatergic innervation.     

The nucleotide metabolism in lactate perfused hearts under ischaemic and reperfused conditions

It was examined whether lactate influences postischaemic hemodynamic recovery as a function of the duration of ischaemia and whether changes in high-energy phosphate metabolism under ischaemic and reperfused conditions could be held responsible for impairment of cardiac function. To this end, isolated working rat hearts were perfused with either glucose (11 mM), glucose (11 mM) plus lactate (5 mM) or glucose (11 mM) plus pyruvate (5 mM). The extent of ischaemic injury was varied by changing the intervals of ischaemia, i.e. 15, 30 and 45 min. Perfusion by lactate evoked marked depression of functional recovery after 30 min of ischaemia. Perfusion by pyruvate resulted in marked decline of cardiac function after 45 min of ischaemia, while in glucose perfused hearts hemodynamic performance was still recovered to some extent after 45 min of ischaemia. Hence, lactate accelerates postischaemic hemodynamic impairment compared to glucose and pyruvate. The marked decline in functional recovery of the lactate perfused hearts cannot be ascribed to the extent of degradation of high-energy phosphates during ischaemia as compared to glucose and pyruvate perfused hearts. Glycolytic ATP formation (evaluated by the rate of lactate production) can neither be responsible for loss of cardiac function in the lactate perfused hearts. Moreover, failure of reenergization during reperfusion, the amount of nucleosides and oxypurines lost or the level of high-energy phosphates at the end of reperfusion cannot explain lactate-induced impairment. Alternatively, the accumulation of endogenous lactate may have contributed to ischaemic damage in the lactate perfused hearts after 30 min of ischaemia as it was higher in the lactate than in the glucose or pyruvate perfused hearts. It cannot be excluded that possible beneficial effects of the elevated glycolytic ATP formation during 15 to 30 min of ischaemia in the lactate perfused hearts are counterbalanced by the detrimental effects of lactate accumulation.     

Excitatory Amino Acid-Evoked Release of γ-[3H]Aminobutyric Acid from Striatal Neurons in Primary Culture

The actions of excitatory amino acids on the release of previously incorporated gamma-[3H]aminobutyric acid ([3H]GABA) were examined in purified (greater than 93%) striatal neurons derived from the fetal mouse brain and differentiated in primary culture. Glutamate, KCl, and veratrine evoked a dose-dependent, saturable, and reversible release of [3H]GABA from striatal neurons. Glutamate actions were not reduced in the absence of calcium, and were insensitive to tetrodotoxin. The dose-response relationships of excitatory amino acids demonstrated the following rank order of potency: glutamate greater than aspartate = N-methyl-D-aspartate greater than kainate much greater than quisqualate. Kainate, however, was the most effective agonist, evoking an eightfold increase over baseline levels of [3H]GABA release. Aspartate- and N-methyl-D-aspartate-evoked release was abolished in the presence of either 2-aminophosphonovaleric acid or gamma-D-glutamylglycine. Release due to glutamate and kainate was partially or ineffectively attenuated by these agents. Glutamate-, aspartate-, and N-methyl-D-aspartate-evoked GABA releases were augmented when calcium was omitted from the bathing medium and reduced when sodium was replaced with choline or lithium. Kainate-evoked release was unaffected when calcium was omitted, virtually unchanged when choline replaced sodium, and markedly potentiated when lithium was substituted for sodium. These findings suggest that at least two distinct receptor systems for excitatory amino acids mediate the evoked release of [3H]GABA from striatal neurons in primary culture. These two systems, aspartate/N-methyl-D-aspartate- and kainate-preferring, are distinguishable on the basis of their pharmacological and ionic properties.     

The effects of acid and acid/aluminum exposure on circulating plasma cortisol levels and other blood parameters in the rainbow trout, Salmo gairdneri

Softwater (Ca²⁺=50, Na⁺= 50(μequiv. l⁻¹) acclimated rainbow trout were fitted with chronic arterial catheters to allow for repetitive blood sampling. After 48 h recovery they were then exposed to either control (pH 6.5, Al = 0μg l⁻¹), acid (pH 4.8, Al = 0μg l⁻¹) or acid plus aluminum (pH 4.8, A1 = 112 μg l⁻¹) conditions for 72 h. Parameters measured included blood glucose, lactate, haemoglobin, haematocrit and plasma Na⁺, Cl⁻, protein and cortisol.
Exposure to pH 4'8 alone caused no mortality, a moderate ionoregulatory disturbance and a transient elevation in plasma cortisol. All other parameters were not significantly different from controls. Addition of aluminum to this exposure caused 100% mortality with a mean survival time of only 27.0 h. There was a marked decrease in plasma ions, hyperglycemia, lactate accumulation, haemoconcentration, red cell swelling, and a sharp rise in plasma cortisol becoming greatly increased as the fish neared death. The mechanism of toxicity of acute acid/aluminum exposure, the role for cortisol under such conditions, and the validity of cortisol and glucose as indicators of stress in fish are discussed.     

Effects of ammonium on energy metabolism and intracellular pH in guinea pig cerebral cortex studied by P and H nuclear magnetic resonance spectroscopy

³¹P and ¹H nuclear magnetic resonance spectroscopy were used to study the effects of ammonium on high-energy phosphates, intracellular pH and lactate in guinea pig cerebral cortex in vitro. In the presence of glucose, 1 mM ammonium caused an intracellular acidification by 0.2–0.3 pH units without a change in phosphocreatine/ATP (PCr/ATP) ratio, lactate concentration or oxygen uptake. At concentrations of 5 mM or greater, NH₄⁺ caused an energy failure and an increase in tissue lactate, together with a drop in intracellular pH. A split in the inorganic phosphate resonance was observed during the exposure to both 20 mM NH₄⁺ and 20 mM K⁺ indicating heterogeneity of the volume-averaged intracellular pH. Cortical brain slices incubated in the presence of 10 mM lactate maintained PCr/ATP ratio and intracellular pH at similar levels as in the presence of glucose, but 1 mM NH₄⁺ caused a fall in PCr/ATP. Both 20 mM NH₄⁺ and 20 mM K⁺ stimulated oxygen uptake of the preparation with glucose or lactate as substrate. These results show that the only acute effect of 1 mM NH₄⁺ in the presence of glucose is an intracellular acidification whereas energetic consequences develop at high levels of this neurotoxic agent.     

Domoic acid, the alleged "mussel toxin," might produce its neurotoxic effect through kainate receptor activation: an electrophysiological study in the dorsal hippocampus

Domoic acid, an excitatory amino acid structurally related to kainate, was recently identified as being presumably responsible for the recent severe intoxication presented by more than 100 people having eaten mussels grown in Prince Edward Island (Canada). The amino acid kainate has been shown to be highly neurotoxic to the hippocampus, which is the most sensitive structure in the central nervous system. The present in vivo electrophysiological studies were undertaken to determine if domoic acid exerts its neurotoxic effect via kainate receptor activation. Unitary extracellular recordings were obtained from pyramidal neurons of the CA1 and the CA3 regions of the rat dorsal hippocampus. The excitatory effect of domoic acid applied by microiontophoresis was compared with that of agonists of the three subtypes of glutamatergic receptors: kainate, quisqualate, and N-methyl-D-aspartate. In CA1, the activation induced by domoic acid was about threefold greater than that induced by kainate; identical concentrations and similar currents were used. In CA3, domoic acid was also three times more potent than kainate. However, the most striking finding was that domoic acid, similar to kainate, was more than 20-fold more potent in the CA3 than in the CA1 region, whereas no such regional difference could be detected with quisqualate and N-methyl-D-aspartate. As the differential regional response of CA1 and CA3 pyramidal neurons to kainate is attributable to the extremely high density of kainate receptors in the CA3 region, these results provide the first electrophysiological evidence that domoic acid may produce its neurotoxic effects through kainate receptor activation.     

Rapid ATP Loss Caused by Methamphetamine in the Mouse Striatum: Relationship Between Energy Impairment and Dopaminergic Neurotoxicity

Abstract: To study the relationship between energy impairment and the effects of α-methamphetamine (METH) on dopaminergic neurons, ATP and dopamine levels were measured in the brain of C57BL/6 mice treated with either a single or four injections of METH (10 mg/kg, i.p.) at 2-h intervals. Neither striatal ATP nor dopamine concentrations changed after a single injection of METH, but both were significantly decreased 1.5 h after the multiple-dose regimen. The effects of METH on ATP levels appear to be selective for the striatum, as ATP concentrations were not affected in the cerebellar cortex and hippocampus after either a single or multiple injections of METH. In a second set of experiments, an intraperitoneal injection of 2-deoxyglucose (2-DG; 1 g/kg), an inhibitor of glucose uptake and utilization, was given 30 min before the third and fourth injections of METH. 2-DG significantly potentiated METH-induced striatal ATP loss at 1.5 h and dopamine depletions at 1.5 h and 1 week. These results indicate that a toxic regimen of METH selectively causes striatal energy impairment and raise the possibility that perturbations of energy metabolism play a role in METH-induced dopaminergic neurotoxicity.     

Immediate and Long-Term Effects of 5,7-Dihydroxytryptamine on Rat Striatal Serotonergic Neurons Measured Using In Vivo Voltammetry

The immediate and long-term effects of the selective serotonergic neurotoxin 5,7-dihydroxytryp-tamine (5,7-DHT) on rat striatal serotonergic neurons were examined after its intracerebroventricular administration using in vivo voltammetry. Extracellular concentration of 5-hydroxyindoles increased immediately following intracerebroventricular 5,7-DHT injection (200 g in 24 l, 18 min), peaked at 1.5-2 h, and returned to normal by 4 h. 5,7-DHT diffused to the contralateral striatum in detectable amounts 9 to 12 min after the start of injection and returned to basal levels by 1.5 h. Three to 6 days after 5,7-DHT lesions, 5-hydroxytryptophan administration produced an increase in striatal 5-hydroxyindoles that was greater than that produced in pre-lesioned rats. This effect was maximal at 14 to 17 days post-lesion, and remained even after 50 days. The short-term effect of 5,7-DHT may be attributable to increased serotonin release, inhibition of uptake, or monoamine oxidase inhibition. The long-term effect of 5,7-DHT lesions may attributable to increased synthesis of serotonin or decreased reuptake in remaining serotonergic neurons.     

Neurons and Neuronal Stem Cells Survive in Glucose-Free Lactate and in High Glucose Cell Culture Medium During Normoxia and Anoxia

Several questions concerning the survival of isolated neurons and neuronal stem and progenitor cells (NPCs) have not been answered in the past: (1) If lactate is discussed as a major physiological substrate of neurons, do neurons and NPCs survive in a glucose-free lactate environment? (2) If elevated levels of glucose are detrimental to neuronal survival during ischemia, do high concentrations of glucose (up to 40 mmol/L) damage neurons and NPCs? (3) Which is the detrimental factor in oxygen glucose deprivation (OGD), lack of oxygen, lack of glucose, or the combination of both? Therefore, in the present study, we exposed rat cortical neurons and NPCs to different concentrations of d-glucose ranging from 0 to 40 mmol/L, or 10 and 20 mmol/L l-lactate under normoxic and anoxic conditions, as well as in OGD. After 24 h, we measured cellular viability by biochemical assays and automated cytochemical morphometry, pH values, bicarbonate, lactate and glucose concentrations in the cell culture media, and caspases activities. We found that (1) neurons and NPCs survived in a glucose-free lactate environment at least up to 24 h, (2) high glucose concentrations >5 mmol/L had no effect on cell viability, and (3) cell viability was reduced in normoxic glucose deprivation to 50% compared to 10 mmol/L glucose, whereas cell viability in OGD did not differ from that in anoxia with lactate which reduced cell viability to 30%. Total caspases activities were increased in the anoxic glucose groups only. Our data indicate that (1) neurons and NPCs can survive with lactate as exclusive metabolic substrate, (2) the viability of isolated neurons and NPCs is not impaired by high glucose concentrations during normoxia or anoxia, and (3) in OGD, low glucose concentrations, but not low oxygen levels are detrimental for neurons and NPCs.     

Biochemical and Neurotoxicological Effects of L-2-Chloropropionic Acid on Rodent Brain

L-2-Chloropropionic acid (L-CPA) is selectively toxic to cerebellar granule cells; necrosis is first observed in rats 36 h after L-CPA administration (750 mg/kg p.o.) and becomes marked by 48 h. L-CPA has also been shown to activate the mitochondrial pyruvate dehydrogenase (PDH) complex in fasted adult rats, resulting in reduced blood glucose and lactate levels. This study aimed to investigate the biochemical and neurotoxicological effects of L-CPA on the brain. Extracts, prepared from guinea-pig cerebellar and cerebral cortex slices incubated in the presence of L-CPA, were analysed using 1H magnetic resonance spectroscopy, 31P magnetic resonance spectroscopy, and amino acid analysis. Glucose metabolism was studied by monitoring the metabolism of [1-(13)C]glucose using gas chromatography/mass spectrometry. Increased glucose metabolism and decreases in the pool sizes of lactate and alanine were observed in both tissues, demonstrating activation of the PDH complex. Extracts were also prepared from the forebrain and cerebellum of animals that had been treated in vivo with L-CPA and analysed as described for the in vitro studies. Similar evidence for PDH activation was demonstrated at 2 and 24 h after dosing in both tissues. At 48 h after dosing, when signs of toxicity are observed, an increase in the lactate concentration and a decrease in N-acetylaspartate in the cerebellum but not in the forebrain confirmed the selective neurotoxic action of L-CPA. These results suggest that activation of the PDH complex does not directly lead to the delayed selective neurotoxicity of L-CPA.     

Effect of lactic acid on growth and butanediol production byKlebsiella oxytoca

Summary Production of 2,3-butanediol byKlebsiella oxytoca was enhanced in the presence of low levels (<8 g/l) of added sodium lactate. Cell growth was inhibited, however, and essentially stopped above 15 g/l added lactate. Levels of by-products (acetic acid and ethanol) were also higher. With 3 g/l lactate and an initial glucose level of 98 g/l, butanediol concentration and productivity increased 164% with 98% utilization of glucose. With high glucose concentration (219 g/l), addition of 2.64 g/l lactate after the growth phase resulted in 81 g/l butanediol, with a productivity of 0.65 g/l/h and 71% glucose utilization.     

Expression of Excitatory Amino Acid Receptors by Cerebellar Cells of the Type-2 Astrocyte Cell Lineage

Abstract: We have used postnatal rat cerebellar astrocyte-enriched cultures to study the excitatory amino acid receptors present on these cells. In the cultures used, type-2 astrocytes (recognized by the monoclonal antibodies A2B5 and LB1) selectively took up γ-[³H]aminobutyric acid ([³H]GABA) and released it when incubated in the presence of micromolar concentrations of kainic and quisqualic acids. The releasing effect of kainic acid was concentration dependent in the range of 5–100 μ M . Quisqualate was more effective than kainate in the lower concentration range but less effective at concentrations at which its releasing activity was maximal (∼50 μ M ). N -Methyl- d -aspartic acid and dihydrokainate (100 μ M ) did not stimulate [³H]GABA release from cultured astrocytes. l -Glutamic acid (20–100 μ M ) stimulated [³H]GABA release as effectively as kainate. The stimulatory effects of kainate and quisqualate on [³H]GABA release were completely Na⁺ dependent; that of kainate was also partially Ca²⁺ dependent. Kynurenic acid (50–200 μ M ) selectively antagonized the releasing effects of kainic acid and also that of l -glutamate; quisqualate was unaffected. Quisqualic acid inhibited the releasing effects of kainic acid when both agonists were used at equimolar concentrations (50 μ M ). d -[³H]aspartate was taken up by both type-1 and type-2 astrocytes, but only type-2 astrocytes released it in the presence of kainic acid. Excitatory amino acid receptors with a pharmacology similar to that of the receptors present in type-2 astrocytes were also expressed by the immature, bipotential progenitors of type-2 astrocytes and oligodendrocytes.     

YM-09151-2 but Not l-Sulpiride Induces Transient Dopamine Release in Rat Striatum via a Tetrodotoxin-Insensitive Mechanism

Abstract: The effects of the selective dopamine D₂ receptor antagonists YM-09151-2 and l -sulpiride on the in vivo release of dopamine (DA), l -3,4-dihydroxyphenylacetic acid (DOPAC), and homovanillic acid (HVA) in rat striatum were investigated. The drugs were injected into the striatum through a microinjection needle attached to a dialysis probe. YM-09151-2 (0.1 or 1.0 μg/0.5 μl) injected into the striatum produced a dramatic rapid-onset transient increase in striatal DA release in a dose-dependent manner. However, the DA increase induced by l -sulpiride (15 or 75 ng/0.5 μl) was small and of slower onset. An increase of DOPAC levels by YM-09151-2 was biphasic: The first peak occurred at 40 min, followed by a delayed-onset gradual increase. Slower-onset gradual increases were also found in DOPAC levels after l -sulpiride injection and in HVA levels after injections of both YM-09151-2 and l -sulpiride. The infusion of tetrodotoxin (TTX; 2 μM) revealed two different types of DA release mechanisms: The rapid-onset transient DA release induced by YM-09151-2 was TTX insensitive, whereas the slower-onset DA release induced by l -sulpiride was TTX sensitive. Moreover, the rapid-onset transient DA release was Ca²⁺ independent and was not affected by pre-treatment with l -sulpiride or nomifensine. Therefore, it is concluded that YM-09151-2 injected into the striatum produced a transient striatal DA release that is independent of D₂ receptors and the action potential.     

Decarboxylation of Exogenous l-3,4-Dihydroxyphenylalanine in Rat Striatum as Studied by In Vivo Voltammetry

An in vivo voltammetric technique was used to determine whether striatal nondopaminergic neurons take up and decarboxylate exogenous L-3,4-dihydroxyphenylalanine (L-DOPA) and release it as dopamine. After the striatal serotonergic neurons of the rat had been destroyed by intraventricular injection of 5,7-dihydroxytryptamine, L-DOPA was administered intraperitoneally. It was found that changes in the dopamine concentration in the striatal extracellular fluid of the rat were the same as those in the nonlesioned rat. L-DOPA was also administered to the rat after the striatal perikarya had been destroyed by the intrastriatal injection of kainate. The striatal dopamine concentrations of the lesioned rat changed in parallel with 5,7-dihydroxytryptamine-lesioned rats, as well as the nonlesioned rats. Moreover, when normal rats were administered L-DOPA, the dopamine concentration was not increased in the cerebellum, where dopamine neurons do not exist. From these observations, it is concluded that exogenous L-DOPA is taken up, decarboxylated to dopamine, and released only in the striatal dopamine neurons.     

Behavioral and biochemical changes following acute administration of MPTP and MPP+

The acute effects of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and 1-methyl-4-phenylpyridinium ion (MPP+) on mouse locomotor activity and striatal dopamine (DA) and 5-hydroxytryptamine (5-HT) levels were investigated. A single dose of either MPTP (10-30 mg/kg, i.p.) or MPP+ (5-20 ug/mouse, i.c.v.) decreased locomotor activity 10-40 min after injection: this locomotor effect was significantly suppressed by either pretreatment with nomifensine or 1-deprenyl alone, or by the combination of desmethylimipramine and 6-hydroxydopamine. Pretreatment with clorgyline did not suppress this behavior and a single dose of haloperidol enhanced the effect. The striatal levels of DA, 3-methoxytyramine and 5-HT increased in parallel with the decrease in locomotor activity caused by MPTP or MPP+. In contrast, levels of 3,4-dihydroxyphenylacetic acid, homovanillic acid and 5-hydroxyindoleacetic acid were decreased by injection of either MPTP or MPP+. Possible mechanism(s) of the behavioral and biochemical changes caused by the acute actions of MPTP and MPP+ with respect to their neurotoxic effects on the nigrostriatal DA system are discussed.     

3-Nitropropionic acid-induced neurotoxicity--assessed by ultra high resolution positron emission tomography with comparison to magnetic resonance spectroscopy

To explore acute and long-term effects of 3-nitropropionic acid (3-NP)-induced neurotoxicity, longitudinal positron emission tomography (PET) studies of energy metabolism and magnetic resonance spectroscopic (MRS) studies of neurochemicals were conducted in a rat model. The first injection of 3-NP (20 mg/kg i.p.) was followed by MRS study of neurochemicals and PET study of glucose utilization using [(18)F]2-fluorodeoxy-D-glucose ((18)F-FDG). After that, 3-NP administration was done two times a day with a dose of 10 mg/kg i.p. until animals were symptomatic or for a maximum of 5 days combined with daily PET studies. Long-term effects were investigated 4 weeks and 4 months after cessation of 3-NP. These studies showed a significant inter-animal variation in response of 3-NP toxicity. Animals that developed large striatal lesions had decreased glucose utilization in the striatum and cortex 1 day after starting 3-NP injections. Similarly succinate and lactate/macromolecule levels were enhanced; these changes being, however, reversible. Progressive degeneration was observed by decreasing striatal glucose utilization and N-acetylaspartate (NAA) and increasing choline. These observations paralleled with weight loss and deficits in behavior. Animals that did not develop lesions showed reversible enhancement in cortical glucose utilization and no change in striatal glucose utilization or neurochemicals or locomotor activity.     

Excitatory Amino Acids: Studies on the Biochemical and Chemical Stability of Ibotenic Acid and Related Compounds

The complex pharmacological profile (excitation/inhibition) of ibotenic acid on single neurons in the mammalian CNS prompted studies on the stability of ibotenic acid and a number of structurally related excitatory amino acids under different in vitro conditions in the presence or absence of enzymes. Ibotenic acid, (RS)-3-hydroxy-4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridine-7-carboxylic acid (7-HPCA), (RS)-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), and (RS)-alpha-amino-3-hydroxy-4-bromo-5-isoxazolepropionic acid (4-Br-homoibotenic acid) were all inhibitors of (S)-glutamic acid decarboxylase (GAD) in mouse brain homogenates, but only ibotenic acid was shown to undergo decarboxylation during incubation with brain homogenates. The formation of the decarboxylated product, muscimol, which primarily occurred in a synaptosomal fraction, was dependent on the presence of pyridoxal-5-phosphate (PALP) and was inhibited by (S)-glutamic acid, 3-mercaptopropionic acid (3MPA), aminooxyacetic acid (AOAA), and allyglycine, suggesting that ibotenic acid is a substrate for GAD. The overall decomposition rate for ibotenic acid (8.7 nmol min-1 mg-1 of protein), which apparently embraces other reactions in addition to decarboxylation to muscimol, was higher than the rate of decarboxylation of (S)-glutamic acid (3.2 nmol min-1 mg-1 of protein). At pH 7.4 and 37 degrees C, but in the absence of enzymes, none of the excitatory amino acids under study underwent any detectable decomposition, whereas ibotenic acid and 7-HPCA, but not AMPA and 4-Br-homoibotenic acid, decomposed, partially by decarboxylation, at 100 degrees C in a pH-dependent manner. In the presence of liver homogenates, ibotenic acid was also shown to decompose.(ABSTRACT TRUNCATED AT 250 WORDS)