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R Schwarcz

Publications and source records attributed to R Schwarcz.

At least 19 recordsLinked to original sources
PubMed1997

Enzyme-catalyzed production of the neuroprotective NMDA receptor antagonist 7-chlorokynurenic acid in the rat brain in vivo.

NMDA receptors play a critical role in neurotransmission and are also involved in the occurrence of excitotoxic nerve cell death. Synthetic halogenated analogs of the endogenous broad spectrum excitatory amino acid receptor blocker kynurenic acid are among the most potent and selective antagonists of the glycine co-agonist site of the NMDA receptor complex. Pharmacological blockade of this site provides neuroprotection in animal models of cerebral ischemia, epilepsy and neurodegenerative disorders, and does not appear to be associated with some of the undesirable side effects linked to classic competitive and non-competitive NMDA receptor antagonists. Here we demonstrate the neuroprotective quantities of 7-chloro-kynurenic acid (7-Cl-KYNA), one of the most selective and well-studied glycine site antagonists, can be synthesized in the brain from its bioprecursor L-4-chlorokynurenine (4-Cl-KYN). Intracerebral infusion of 4-Cl-KYN dose-dependently reduced quinolinate neurotoxicity in the rat hippocampus after enzymatic conversion to 7-Cl-KYNA by kynurenine aminotransferase. In accordance with previous studies demonstrating that kynurenine aminotransferase is preferentially localized in astrocytes, both the enzymatic formation of 7-Cl-KYNA and the neuroprotective potency of 4-Cl-KYN were substantially reduced following an intrahippocampal injection of the gliotoxin fluorocitrate. In situ produced 7-Cl-KYNA offers a novel neuroprotective strategy for targeting the glycine/NMDA site while avoiding excessive receptor blockade and reducing the clinical risks associated with conventional NMDA receptor antagonism.

Animals↗
PubMed1996

Facilitated brain uptake of 4-chlorokynurenine and conversion to 7-chlorokynurenic acid.

7-Chlorokynurenic acid (7-Cl-KYNA) and 5,7-dichlorokynurenic acid (5,7-Cl2-KYNA) are of therapeutic interest as potent glycine/N-methyl-D-aspartate NMDA) receptor antagonists, but are excluded from brain by the blood-brain barrier. We examined whether these compounds could be delivered to brain through their respective precursors, L-4-chlorokynurenine (4-Cl-KYN) and L-4,6-dichlorokynurenine (4,6-Cl2-KYN), which are amino acids. 4-Cl-KYN was shown to be rapidly shuttled into the brain by the large neutral amino acid transporter of the blood-brain barrier (K(m) = 105 +/- 14 microM, Vmax = 16.9 +/- 2.3 nmol min-1 g-1) and to be converted intracerebrally to 7-Cl-KYNA. 4,6-Cl2-KYN also expressed affinity for the transporter, but four-fold less than that of 4-Cl-KYN. In summary, the results show that because of their facilitated uptake 4-Cl-KYN and 4,6-Cl2KYN might be useful prodrugs for brain delivery of glycine-NMDA receptor antagonists.

Amino Acids↗
PubMed1996

Characterization of L-alpha-aminoadipic acid transport in cultured rat astrocytes.

The mechanism of the selective gliotoxicity of L-alpha-aminoadipate (L-alpha AA) is thought to involve its entry into glia as a substrate for glutamate transporters or, alternatively, its ability to inhibit glial glutamate transport. To clarify the properties of L-alpha AA as a transport substrate, we explored the ionic dependence, kinetics and pharmacology of L-[3H] alpha AA uptake in rat cortical astrocytes. We observed two components of saturable L-alpha AA uptake, one Na(+)-dependent and the other Na(+)-independent. These components exhibited the characteristics of system X-AG, the widespread family of Na(+)-cotransporters of aspartate and glutamate, and system x-c, a Cl(-)-dependent glutamate/cystine exchanger, respectively. The K(m) value of Na(+)-dependent L-alpha AA uptake was 629 +/- 42 microM, and Vmax was 62 +/- 4 nmol.min-1.mg-1 protein, which was more than twice the capacity of Na(+)-dependent glutamate uptake. The kinetic parameters of Na(+)-dependent L-alpha AA uptake (K(m) of 20 +/- 2 microM, Vmax of 1.7 +/- 0.4 nmol.min-1.mg-1 protein did not differ from the values for Na(+)-independent glutamate uptake, indicating that L-alpha AA and glutamate are equally good substrates for system x-c.

2-Aminoadipic Acid↗
PubMed1996

(R,S)-3,4-dichlorobenzoylalanine (FCE 28833A) causes a large and persistent increase in brain kynurenic acid levels in rats.

Kynurenic acid is an endogenous excitatory amino-acid receptor antagonist with neuroprotective and anticonvulsant properties. We demonstrate here that systemic administration of the new and potent kynurenine 3-hydroxylase inhibitor (R,S)-3,4-dichlorobenzoylalanine (FCE 28833A) causes a dose-dependent elevation in endogenous kynurenine and kynurenic acid levels in rat brain tissue. In hippocampal microdialysates, peak increases of 10- and 80-fold above basal kynurenic acid concentrations, respectively, were obtained after a single oral or intraperitoneal administration of 400 mg/kg FCE 28833A. After intraperitoneal treatment with FCE 28833A, extracellular brain kynurenic acid levels remained significantly elevated for at least 22 h, rendering this compound a far more effective enhancer of kynurenic acid levels than the previously described kynurenine 3-hydroxylase blocker m-nitrobenzoylalanine. FCE 28833A and similar molecules may have therapeutic value in diseases which are linked to a hyperfunction of excitatory amino-acid receptors.

Animals↗
PubMed1996

Study of receptor-mediated neurotoxins released by HIV-1-infected mononuclear phagocytes found in human brain.

Although there is growing evidence that neurotoxic molecules produced by HIV-1-infected mononuclear phagocytes damage neurons, the precise mechanisms of neuronal attack remain uncertain. One class of cytotoxin involves neuronal injury mediated via the NMDA receptor. We examined blood monocytes and brain mononuclear cells isolated at autopsy from HIV-1-infected individuals for the ability to release NMDA-like neuron-killing factors. We found that a neurotoxic amine, NTox, was produced by blood monocytes and by brain mononuclear phagocytes infected with retrovirus. In vivo injections of minute quantities of NTox produced selective damage to hippocampal pyramidal neurons. NTox can be extracted directly from brain tissues infected with HIV-1 and showed structural features similar to wasp and spider venoms. In contrast to NTox, HIV-1 infection did not increase the release of the NMDA excitotoxin quinolinic acid (QUIN) from mononuclear cells. Although we found modest elevations of QUIN in the CSF of HIV-1-infected individuals, the increases were likely attributable to entry through damaged blood-brain barrier. Taken together, our data pinpoint NTox, rather than QUIN, as a major NMDA receptor-directed toxin associated with neuro-AIDS.

Acquired Immunodeficiency Syndrome↗
PubMed1996

Excitotoxic lesions of the rat striatum: different responses of kynurenine pathway enzymes during ontogeny.

Excitotoxic lesions of the adult rat striatum result in reactive gliosis and an associated increase in the activities of the astrocytic enzymes 3-hydroxyanthranilic acid oxygenase (3HAO) and kynurenine aminotransferase (KAT), which are responsible for the biosynthesis of the neurotoxin quinolinic acid and the neuroprotectant kynurenic acid, respectively. Unilateral ibotenate injections were made in the striatum of 7-, 14-, 21- and 28-day- and 2.5-month-old rats to study the reaction of 3HAO and KAT when injury is inflicted during ontogeny. By one week, all lesioned striata showed a > 50 percent decrease in the activity of the neuronal marker enzyme glutamic acid decarboxylase. At this timepoint, lesion-induced elevations in 3HA0 activity increased progressively from 130 to 206, 280, 385 and 456 percent of the contralateral striatum in the five age groups studied. In contrast, in the same animals the respective increases in striatal KAT activity were 601, 350, 312, 259 and 159 percent (n = 6-13 per group). In all age groups, statistically significant lesion-induced increases in 3HA0 and KAT were seen up to 4 weeks after the ibotenate injection. Rats receiving an intrastriatal injection of ibotenate on postnatal day 7 also showed an increase in the striatal tissue level of kynurenic acid 1 week after the lesion. These data demonstrate that substantial qualitative differences exist between the immature and adult rat in the reaction of two glial enzymes to striatal injury. Moreover, the ability of the immature brain to mobilize kynurenic acid production preferentially may play a role in the brain's response to perinatal injury.

3-Hydroxyanthranilate 3,4-Dioxygenase↗
PubMed1996

Afferents to the seizure-sensitive neurons in layer III of the medial entorhinal area: a tracing study in the rat.

Neurons in layer III of the medial entorhinal area (MEA) in the rat are extremely vulnerable to local injections of amino-oxyacetic acid and to experimentally induced limbic seizures. A comparable specific pathology has been noted in surgical specimens from patients with temporal lobe epilepsy. Efforts to understand this preferential neuronal vulnerability led us to study the neural input to this layer in the rat. Iontophoretic injection of the retrograde tracer fast blue, aimed at layer III of the MEA, resulted in retrogradely labeled neurons in the presubiculum in all the injected hemispheres. The nucleus reuniens thalami, the anteromedial thalamic nucleus, the ventral portion of the claustrum (endopiriform nucleus), the dorsomedial parts of the anteroventral thalamic nucleus, and the septum-diagonal band complex were labeled less frequently. In only one experiment, retrogradely labeled neurons were observed in the ventrolateral hypothalamus and in the brainstem nucleus raphe dorsalis. Since projections from claustrum to the entorhinal cortex has not been studied in the rat with modern sensitive anterograde tracing techniques, iontophoretic injections of the anterograde tracer Phaseolus vulgaris-leucoagglutinin were placed into the ventral portion of the claustrum. Anterogradely labeled fibers in the entorhinal area proved not to be confined to the MEA, since a prominent projection distributed to the lateral entorhinal area as well. In both areas, the densest terminal labeling was present in layers IV-VI, whereas layer III appeared to be only sparsely labeled. The present data indicate that of all potential afferents only those from the presubiculum distribute preferentially to layer III of the MEA. This, in turn, suggests a potentially important role of the presubiculum in the seizure-related degeneration of neurons in layer III of the MEA.

Afferent Pathways↗
PubMed1996

Seizure activity causes elevation of endogenous extracellular kynurenic acid in the rat brain.

This study was designed to examine the effects of several classic convulsants on the extracellular concentration of the anticonvulsant and neuroprotective brain metabolite kynurenic acid (KYNA) in the rat brain. Drug effects were investigated in vivo, mostly by unilateral microdialysis in the dorsal hippocampus. Systemic administration of pentylenetetrazole (60 mg/kg, SC), pilocarpine (325 mg/kg, SC), bicuculline (6 mg/kg, SC), or kainic acid (10 mg/kg, SC) caused characteristic clonic and/or tonic convulsions. In all seizure paradigms, KYNA levels in the dialysate began to rise within 1 h and gradually reached a plateau approximately 4 h after administration of the convulsants. Peak increases were 1.5-3-fold over basal levels. The duration of the elevation in KYNA levels was significantly prolonged following kainic acid application. In the kainic acid model, extracellular KYNA was also measured and found to be increased in the ventral hippocampus, piriform cortex, and striatum. Moreover, temporary intrahippocampal infusion of the KYN synthesis inhibitor aminooxyacetic acid (1 mM) in the kainic acid- and pentylenetetrazole models attenuated the increase in extracellular KYNA levels, demonstrating that de novo production of KYNA in the brain accounts for the seizure-induced KYNA overflow. A separate group of animals received a unilateral intrahippocampal injection of the endogenous convulsant excitotoxin quinolinic acid (120 nmol) and showed long-lasting (> 24 h) bilateral increases in extracellular KYNA levels. Taken together, these data indicate that an increase in extracellular KYNA may constitute a common occurrence in response to seizures and that KYNA elevations may signify the brain's attempt to counteract seizure activity.

Animals↗
PubMed1995

Electrical kindling is associated with a lasting increase in the extracellular levels of kynurenic acid in the rat hippocampus.

Endogenous kynurenic acid (KYNA), an excitatory amino acid receptor antagonist with antineurotoxic and anticonvulsant activity, was assessed by microdialysis in the hippocampus of kindled rats. One week after the completion of amygdala or hippocampal kindling (stage 5), the dialysate concentration of KYNA in the hippocampus of both hemispheres was 1.7 +/- 0.1-fold higher than in shams (P < 0.01). Veratridine (50 microM), applied through the probe, reduced extracellular KYNA by 28% within 1 h in controls (P < 0.05), but was ineffective in stage 5 kindled rats. At the preconvulsive stage 2, dialysate KYNA concentration and the effect of veratridine were similar to controls. The activity of KYNA's biosynthetic enzyme, kynurenine aminotransferase, did not change in the hippocampus 1 week after stage 5 seizures. These data indicate an enhanced liberation of KYNA in teh hippocampus of fully kindled animals due to an impairment of normal regulatory mechanisms. This may be of relevance for the control of hippocampal excitability during epileptogenesis.

Animals↗
PubMed1995

L-alpha-aminoadipic acid as a regulator of kynurenic acid production in the hippocampus: a microdialysis study in freely moving rats.

L-alpha-Aminoadipic acid is a lysine metabolite with neuroexcitatory properties, and has previously been shown to inhibit the production of the broad spectrum excitatory amino acid receptor antagonist kynurenic acid in brain tissue slices. The effects of L-alpha-aminoadipic acid on the levels of extracellular kynurenic acid were now studied by microdialysis in the dorsal hippocampus of freely moving rats. Application of L-alpha-aminoadipic acid through the microdialysis probe dose dependently decreased both the concentration of endogenous kynurenic acid and of kynurenic acid which was produced de novo from its bioprecursor L-kynurenine (500 microM applied through the probe). 500 microM L-alpha-aminoadipic acid lowered the kynurenic acid concentration in the dialysate by 47% and 28% with and without precursor loading, respectively, whereas D-alpha-aminoadipic acid was without effect. Co-administration of 500 microM L-alpha-aminoadipic acid with 50 microM veratridine, which by itself produces a substantial decrease in the levels of extracellular kynurenic acid, did not result in a further reduction in kynurenic acid concentrations. Extensive neuronal degeneration caused by an intrahippocampal injection of quinolinic acid (120 nmol) did not interfere with the effect of L-alpha-aminoadipic acid. Taken together, these data suggest that the effect of L-alpha-aminoadipic acid on extracellular kynurenic acid levels is likely due to its direct action on astrocytes, which are known to harbor kynurenic acid's biosynthetic enzyme, kynurenine aminotransferase. L-alpha-Aminoadipic acid may modulate kynurenic acid function in the brain and thus play a role in the pathogenesis of neurodegenerative and seizure disorders.

2-Aminoadipic Acid↗
PubMed1995

Dysfunction of brain kynurenic acid metabolism in Huntington's disease: focus on kynurenine aminotransferases.

The levels of the neuroprotective excitatory amino acid receptor antagonist kynurenic acid (KYNA) have been previously shown to be reduced in several regions of the brain of Huntington's disease (HD) patients. Thus, KYNA has been speculatively linked to the pathogenesis of HD. We have examined KYNA levels and the activity of its two biosynthetic enzymes (kynurenine aminotransferases (KAT) I and II) in 12 regions of brains from late-stage HD patients and control donors (n = 17 each). KYNA levels were measured in the original tissue homogenate. Using [3H]kynurenine as the substrate, enzyme activities were determined in dialyzed tissue homogenates. KYNA levels in the caudate nucleus decreased from 733 +/- 95 in controls to 401 +/- 62 fmol/mg tissue in HD (p < 0.01). The activity of both enzymes was highest in cortical areas (e.g. control frontal cortex: KAT I: 148 +/- 18 fmol/mg tissue/h; KAT II: 25 +/- 2 fmol/mg tissue/h). The activities of both KAT I and KAT II, when expressed per mg original weight, showed significant decreases (48-55%) in the HD putamen (p < 0.01). Trends toward lower enzyme activities and KYNA concentrations were detected in other brain areas as well. Kinetic analyses, performed in putamen and cerebellum, showed an approximately 3-fold increase in Km values for both KAT I and KAT II in the putamen only. Vmax values remained unchanged in the HD brain. These findings indicate a selective impairment in KYNA biosynthesis in the neostriatum of HD patients, possibly due to the loss of (an) endogenous KAT activator(s).(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗
PubMed1995

Differential neuronal vulnerability to amino-oxyacetate and quinolinate in the rat parahippocampal region.

Injection of the "indirect" excitotoxin amino-oxyacetate into the entorhinal area causes acute behavioral seizures and preferential neuronal loss in layer III of the medial entorhinal cortex in rats. We examined here whether the effects of amino-oxyacetate could be duplicated by local injections of the endogenous N-methyl-D-aspartate receptor agonist and direct excitotoxin, quinolinate. Amino-oxyacetate (685 nmol) or quinolinate (30, 45 or 60 nmol) were injected into the entorhinal cortex of rats anesthetized with choral hydrate (360 mg/kg). Separate groups of animals were co-treated with the N-methyl-D-aspartate receptor antagonist dizocilpine maleate (2 mg/kg) or given a higher dose of chloral hydrate (500 mg/kg). Rats that received amino-oxyacetate and a low anesthetic dose consistently displayed acute behavioral seizures and showed preferential loss of neurons in layer III of the medial entorhinal cortex. Animals that were given quinolinate did not display behavioral seizures, and showed preferential degeneration of neurons in layer V of the entorhinal cortex. Moreover, quinolinate-injected rats frequently exhibited neuronal loss in the superficial layers of the dorsal perirhinal cortex. The behavioral and neuropathological sequelae of amino-oxyacetate, but not quinolinate-induced neurotoxicity, were abolished by prolonged chloral hydrate anesthesia. In spite of these apparent qualitative differences between the two toxins, neurodegeneration induced by either amino-oxyacetate or quinolinate was completely prevented by dizocilpine maleate. These data suggest that a heterogeneous distribution of pharmacologically distinct N-methyl-D-aspartate receptor subtypes in the parahippocampal region may underlie the distinct neurodegenerative properties of the two toxins. Since the lesion caused by amino-oxyacetate bears remarkable similarities to neuropathological changes which have been described in this structure in temporal lobe epilepsy, further elucidation of the mechanisms of cellular toxicity of amino-oxyacetate may hold clues for the pathogenesis of this disease.

Aminooxyacetic Acid↗