Effects Of Kynurenic Acid Research Articles

P2-297 Differential and isoform-specific effects of apolipoprotein E4 on the initiation of deposition and on the reversible dissolution and fibrillization of Ab in vivo

Kynurenic acid (KYNA), one of the main product of the kynurenine pathway originating from tryptophan, is considered to be neuroprotective. Dysregulation of KYNA activity is thought to be involved in neurodegenerative diseases, the physiopathology of which evokes excitotoxicity, oxidative stress and/or protein aggregation. The neuroprotective effect of KYNA is generally attributed to its antagonistic action on NMDA receptors. However, this single target action appears insufficient to support KYNA beneficial effects against complex neurodegenerative processes including neuroinflammation, β-amyloid peptide (Aβ) toxicity and apoptosis. Novel insights are therefore required to elucidate KYNA neuroprotective mechanisms. Here, we combined cellular, biochemical, molecular and pharmacological approaches to demonstrate that low micromolar concentrations of KYNA strongly induce neprilysin (NEP) gene expression, protein level and enzymatic activity increase in human neuroblastoma SH-SY5Y cells. Furthermore, our studies revealed that KYNA exerts a protective effect on SH-SY5Y cells by increasing their viability through a mechanism independent from NMDA receptors. Interestingly, KYNA also induced NEP activity and neuroprotection in mouse cortical neuron cultures the viability of which was more promoted than SH-SY5Y cell survival under KYNA treatment. KYNA-evoked neuroprotection disappeared in the presence of thiorphan, an inhibitor of NEP activity. NEP is a well characterized metallopeptidase whose deregulation leads to cerebral Aβ accumulation and neuronal death in Alzheimer's disease. Therefore, our results suggest that a part of the neuroprotective role of KYNA may depend on its ability to induce the expression and/or activity of the amyloid-degrading enzyme NEP in nerve cells.

Comparative study on the effects of kynurenic acid and glucosamine–kynurenic acid

Kynurenic acid (KYNA) is the only known endogenous N-methyl- d-aspartate (NMDA) receptor inhibitor and might therefore come into consideration as a therapeutic agent in certain neurobiological disorders. However, its use as a neuroprotective compound is practically excluded because KYNA does not readily cross the blood-brain barrier (BBB). We recently synthetized a new compound, glucosamine–kynurenic acid (KYNA-NH-GLUC), which is presumed to cross the BBB more easily. In this study, the effects of KYNA and KYNA-NH-GLUC on behavior and cortical activity were investigated in adult rats. The results show that (1) on intracerebroventricular application, the behavioral changes induced by KYNA and by KYNA-NH-GLUC are quite similar; (2) on intravenous administration, KYNA (25 mg/kg) has no effect on the somatosensory-evoked cortical potentials, whereas KYNA-NH-GLUC (25 mg/kg) causes transient but appreciable reductions in the amplitudes of the evoked responses within 5 min after application; and (3) the results of in vitro studies demonstrated that both KYNA and KYNA-NH-GLUC reduced the amplitudes of the field excitatory postsynaptic potentials (fEPSPs). These observations suggest that the two compounds have similar effects, but that KYNA-NH-GLUC passes the BBB much more readily than does KYNA. These results imply that the conjugated NH-GLUC is of importance in the passage across the BBB.

Effects of kynurenic acid as a glutamate receptor antagonist in the guinea pig.

Glutamate excitotoxicity is implicated in both the genesis of neural injury and noise-induced hearing loss (NIHL). Acoustic overstimulation may result in excessive synaptic glutamate, resulting in excessive binding to post-synaptic receptors and the initiation of a destructive cascade of cellular events, thus leading to neuronal degeneration and NIHL. The purpose of this study was to determine whether this apparent excitotoxicity can be attenuated by kynurenic acid (KYNA), a broad-spectrum glutamate receptor antagonist, and protect against noise-induced temporary threshold shifts (TTS). Guinea pigs were randomly assigned to three separate groups. Base-line compound action potentials (CAP) thresholds and cochlear microphonics (CM) were recorded. Group I was treated with physiologic saline as a vehicle control applied to the round window membrane that was followed by 110 dB SPL wide-band noise for 90 min. Group II received 5 mM KYNA followed by noise exposure, and group III received 5 mM KYNA alone without noise exposure. Post-drug and noise levels of CAP thresholds and CM were then obtained. Noise exposure in the control group caused a significant temporary threshold shift (TTS) of 30-40 dB across the frequencies tested (from 3 kHz to 18 kHz). Animals that received 5 mM KYNA prior to noise exposure (group II) showed statistically significant protection against noise-induced damage and demonstrated a minimal TTS ranging between 5 and 10 dB at the same frequencies. Animals in group III receiving KYNA without noise exposure showed no change in thresholds. Additionally, cochlear microphonics showed no considerable difference in threshold shifts when controls were compared to KYNA-treated animals. These results show that antagonizing glutamate receptors can attenuate noise-induced TTS, suggesting that glutamate excitotoxicity may play a role in acoustic trauma.

Respiratory effects of kynurenic acid microinjected into the ventromedullary surface of the rat.

Several studies demonstrate that, within the ventral medullary surface (VMS), excitatory amino acids are necessary components of the neural circuits involved in the tonic and reflex control of respiration and circulation. In the present study we investigated the cardiorespiratory effects of unilateral microinjections of the broad spectrum glutamate antagonist kynurenic acid (2 nmol/200 nl) along the VMS of urethane-anesthetized rats. Within the VMS only one region was responsive to this drug. This area includes most of the intermediate respiratory area, partially overlapping the rostral ventrolateral medulla (IA/RVL). When microinjected into the IA/RVL, kynurenic acid produced a respiratory depression, without changes in mean arterial pressure or heart rate. The respiratory depression observed was characterized by a decrease in ventilation, tidal volume and mean inspiratory flow and an increase in respiratory frequency. Therefore, the observed respiratory depression was entirely due to a reduction in the inspiratory drive. Microinjections of vehicle (200 nl of saline) into this area produced no significant changes in breathing pattern, blood pressure or heart rate. Respiratory depression in response to the blockade of glutamatergic receptors inside the rostral VMS suggests that neurons at this site have an endogenous glutamatergic input controlling the respiratory cycle duration and the inspiratory drive transmission.

3-Nitropropionic acid (3-NPA)-induced toxicity of endothelial cells in vitro is mediated by nitric oxide dependent and independent mechanisms

Kynurenic acid (KYNA) is an endogenous metabolite of the kynurenine pathway for tryptophan degradation and an antagonist of both N-methyl-d-aspartate (NMDA) and alpha-7 nicotinic acetylcholine (α7nACh) receptors. KYNA has also been shown to scavenge hydroxyl radicals (OH) under controlled conditions of free radical production. In this work we evaluated the ability of KYNA to scavenge superoxide anion (O2−) and peroxynitrite (ONOO−). The scavenging ability of KYNA (expressed as IC50 values) was as follows: OH = O2− > ONOO−. In parallel, the antiperoxidative and scavenging capacities of KYNA (0–150 μM) were tested in cerebellum and forebrain homogenates exposed to 5 μM FeSO4 and 2.5 mM 3-nitropropionic acid (3-NPA). Both FeSO4 and 3-NPA increased lipid peroxidation (LP) and ROS formation in a significant manner in these preparations, whereas KYNA significantly reduced these markers. Reactive oxygen species (ROS) formation were determined in the presence of FeSO4 and/or KYNA (0–100 μM), both at intra and extracellular levels. An increase in ROS formation was induced by FeSO4 in forebrain and cerebellum in a time-dependent manner, and KYNA reduced this effect in a concentration-dependent manner. To further know whether the effect of KYNA on oxidative stress is independent of NMDA and nicotinic receptors, we also tested KYNA (0–100 μM) in a biological preparation free of these receptors – defolliculated Xenopus laevis oocytes – incubated with FeSO4 for 1 h. A 3-fold increase in LP and a 2-fold increase in ROS formation were seen after exposure to FeSO4, whereas KYNA attenuated these effects in a concentration-dependent manner. In addition, the in vivo formation of OH evoked by an acute infusion of FeSO4 (100 μM) in the rat striatum was estimated by microdialysis and challenged by a topic infusion of KYNA (1 μM). FeSO4 increased the striatal OH production, while KYNA mitigated this effect. Altogether, these data strongly suggest that KYNA, in addition to be a well-known antagonist acting on nicotinic and NMDA receptors, can be considered as a potential endogenous antioxidant.

The WAG/Rij Rat Model for Nonconvulsive Absence Epilepsy: Involvement of NonNMDA Receptors

The involvement of AMPA and kainate receptors in nonconvulsive epilepsy was studied by intracerebroventricular injections of AMP A, GDEE, kainic acid and kynurenic acid in WAG/Rij rats. The WAG/Rij rat strain is recognized as an animal model for human absence epilepsy. EEG registrations showed that AMPA (0.1 pmol/5 μl; 1 pmol/5 μl; 10 pmol/5 μl) dose-dependently increased the nonconvulsive absence epilepsy while GDEE (0.2 μmol/5 μl; 1 μmol/5 μl; 5 umol/5 μl) caused a dose-dependent decrease. All effects of GDEE could be blocked by an inactive AMPA dosage. Kainic acid (0.01 nmol/5 μl; 0.1 nmol/5 μl; 0.15 nmol/5 μl) had no effects on the nonconvulsive epilepsy but induced convulsions in the two highest dosages. Kynurenic acid (50 nmol/5 μl; 100 nmol/5 μl; 500 nmol/5 μl) decreased dose-dependently the incidence of nonconvulsive epilepsy. The effect of kynurenic acid could be blocked by a nonconvulsive dosage of kainic acid. These results show that the AMPA and kainate receptor appear to be involved in nonconvulsive epilepsy. Furthermore, blockage of these two receptor subtypes led to an antiepileptic effect without inducing behavioural alterations. Therefore, selective AMPA and kainate receptor antagonists might be potent antiepileptics.

NMDA receptor antagonists that bind to the strychnine-insensitive glycine site and inhibit NMDA-induced Ca 2+ fluxes and [ 3H]GABA release

We have examined the actions of putative antagonists of the strychnine-insensitive glycine-mediated modulation of the N-methyl-D-aspartate (NMDA) receptor using [ 3H]MK801 binding, Ca 2+ influx and [ 3H]GABA release assays. Kynurenic acid and HA-966 inhibited [ 3H]MK801 binding, NMDA and glycine induced Ca 2+ influx measured using fura-2 and NMDA and glycine simulated [ 3H]GABA release. The effects of kynurenic acid could be partially overcome by the addition of excess glutamate and glycine, indicating limited selectivity for the glycine binding site. In addition, a component of the action of kynurenic acid was insensitive to agonist concentration, indicating a third action of kynurenic acid at high concentrations. In contrast, HA-966 was 100-foled selective for the glycine compared to the NMDA site. HA-966 only partially inhibited [ 3H]MK801 binding (IC 50 19.7 μM), NMDA-induced Ca 2+ influx and neurotransmitter release. The failure of HA-966 to completely block NMDA responses, even at high concentrations, suggests that glycine may not be an absolute requirement for the activation of NMDA receptors under these experimental conditions.

Effects of kynurenic acid and ketamine on neonatal sleep in rats.

Kynurenic acid and ketamine are, besides non-specific actions, antagonists of excitatory acidic amino acids in the rat brain. At intraperitoneal doses of 300 mg/kg (1.6 mmol/l) and 5 mg/kg (18 mmol/l), respectively, they are equipotent antagonists of N-methyl-D-aspartate receptors. We studied the acute effects of ketamine and kynurenic acid, at these doses, on sleep patterns of rats during the second and third postnatal weeks with the so called static charge sensitive mattress (SCSB). Both kynurenic acid and ketamine decreased active sleep; ketamine additionally increased quiet state and kynurenic acid increased waking. The decrease in active sleep may be related to glutaminergic NMDA and serotonergic responses in the rat brain but non-specific actions and interactions with other transmitter systems are also involved.

Dyskinesia in the primate following injection of an excitatory amino acid antagonist into the medial segment of the globus pallidus

Injection of an excitatory amino acid antagonist, kynurenic acid, into the medial segment of the globus pallidus of the conscious monkey elicited dyskinesia of the contralateral limbs. In most respects the dyskinesia was indistinguishable from the disorder that is produced by ablation of the subthalamic nucleus, or injection of a GABA antagonist into the subthalamic nucleus. Injections of kynurenic acid into the lateral segment of the globus pallidus, by contrast, did not provoked dyskinesia. The effect of kynurenic acid is attributed to the blockade of neurotransmission from the subthalamic nucleus to the medial pallidal segment, and the results suggest that the neurotransmitter utilised by this pathway is an excitatory amino acid.

The effects of kynurenic acid, quinolinic acid and other metabolites of tryptophan on the development of the high pressure neurological syndrome in the rat

The effects of some biologically active metabolites of tryptophan on the high pressure neurological syndrome (HPNS) were studied. Kynurenic acid, quinolinic acid, 5-hydroxytryptophan, kynurenine and 3-hydroxyanthranilic acid, at doses within the physiological range, were administered exogenously to rats prior to exposure to increased pressure and any effects on the tremor, myoclonus and convulsion end points of the high pressure neurological syndrome were observed. Quinolinic acid (25 and 50 mg/kg) and kynurenine (50 mg/kg) reduced the onset pressure for tremor, but not myoclonus or convulsions. Kynurenic acid (100mg/kg) increased tremor onset pressure; 5-hydroxytryptophan (20 mg/kg) slightly increased onset pressure for tremor but decreased that for myoclonus. 3-Hydroxyanthranilic acid (20 mg/kg) had no significant effect on any of the motor signs of the syndrome. These data provide further support for the idea that the motor events seen in the high pressure neurological syndrome are not produced by a single mechanism. Differences between the responses to related metabolites suggest that the precise balance between compounds such as kynurenic acid and quinolinic acid may be important in the appearance of the high pressure neurological syndrome.

Effects of kynurenic acid on amygdaloid kindling in the rat

The anticonvulsant properties of the endogenous excitatory amino acid antagonist, kynurenic acid (KYA), were studied in prepubescent and adult rats using the amygdaloid kindling model of epilepsy. Treatment with intracerebroventricular KYA (360 nmoles (adult dose) or 240 nmoles (prepubescent dose)) prior to administration of the electrical kindling stimulus significantly reduced the rate of kindling in both age groups. However, there was no significant difference between the KYA-treated and the controls in mean afterdischarge threshold in either age group. The results of this study indicate that with the kindling model KYA has a significant anti-convulsant effect in both prepubescent and adult rats.

Effects of kynurenic acid on evoked extracellular field potentials in the rat olfactory bulb in vivo

The effects of excitatory amino acid antagonists on extracellular field potentials in the olfactory bulb produced by lateral olfactory tract stimulation were analyzed in vivo. The two compounds tested, d-2-amino-5-phosphonovalerate and kynurenic acid, were administered by brain dialysis. Only kynurenic acid was able to effectively suppress the monosynaptic excitation of the major class of interneurons, the granule cells. This pharmacological profile suggests the involvement of non-NMDA receptors.

Non- N-methyl- d-aspartate receptors mediating synaptic transmission in the avian cochlear nucleus: Effects of kynurenic acid, dipicolinic acid and streptomycin

We have examined the effects of a number of excitatory amino acid antagonists on transmission at the cochlear nerve-nucleus magnocellularis synapse in the chicken. Using an in vitro preparation and bath application of drugs, we studied the effects of kynurenic acid and several related substances, streptomycin and a selective N-methyl- d-aspartate receptor antagonist, dl-α-aminosuberate. The last compound had no effect on evoked transmission. Of the various kynurenic acid-related compounds tested, only kynurenic and dipicolinic acid selectively altered responses in nucleus magnocellularis. Quinolinic acid, a kynurenic acid analogue that is structurally akin to dipicolinic acid but which acts selectively at N-methyl- d-aspartate receptors, was without effect. The effect of kynurenic acid was solely inhibitory, completely blocking postsynaptic responses with a potency dependent on the frequency of nerve stimulation. No such frequency dependence was seen with dipicolinic acid although this compound also completely suppressed evoked responses. In addition dipicolinic acid potentiated postsynaptic responses at concentrations only slightly lower than those causing inhibition. Streptomycin inhibited responses in nucleus magnocellularis but this effect seems to result partially from the ability of the drug to inhibit presynaptic calcium influx. Our finding that selective antagonists of N-methyl- d-aspartate receptors were ineffective while antagonists of both receptor types, such as kynurenic and dipicolinic acids, inhibited evoked responses reinforces the conclusion that postsynaptic receptors mediating transmission at this synapse are of thenon- N-methyl- d-aspartate type[Nemeth et al. (1983) Neurosci. Lett. 40, 39–44]. However, differences in the effects of kynurenic and dipicolinic acids on evoked responses indicate that they do not have a single common mechanism of action. Our results with streptomycin, although possibly reflecting the involvement of quisqualate-preferring receptors in transmission, also indicate that the usefulness of this drug as a probe for that receptor subtype may be limited by its non-specific actions. Finally, our findings differ from those in the cat anteroventral cochlear nucleus, mammalian homologue of nucleus magnocellularis, where N-methyl- d-aspartate receptors appear to mediate transmission. This difference between otherwise very similar synapses in two different species suggests that selection of a particular receptor and/or transmitter is largely independent of other functional properties.

The antagonistic effects of kynurenic acid on visually- and acidic amino acid-induced excitation in the cat visual cortex

The antagonistic effects of kynurenic acid on visually- and acidic amino acid-induced excitation in the cat visual cortex