6-cyano-7-nitroquinoxaline-2-3-dione and homocysteic-acid

A brief exposure of hippocampal slices to L-quisqualic acid (QUIS) sensitizes CA1 pyramidal neurons 30- to 250-fold to depolarization by certain excitatory amino acids analogues, e.g., L-2-amino-6-phosphonohexanoic acid (L-AP6), and by the endogenous compound, L-cystine. This phenomenon has been termed QUIS sensitization. A mechanism similar to that previously described for QUIS neurotoxicity has been proposed to describe QUIS sensitization. Specifically, QUIS has been shown to be sequestered into GABAergic interneurons by the System x(c)(-) and subsequently released by heteroexchange with cystine or L-AP6, resulting in activation of non-NMDA receptors. We now report two additional neurotoxins, the Lathyrus excitotoxin, beta-N-oxalyl-L-alpha,beta-diaminopropionic acid (ODAP), and the endogenous compound, L-homocysteic acid (HCA), sensitize CA1 hippocampal neurons >50-fold to L-AP6 and >10-fold to cystine in a manner similar to QUIS. While the cystine- or L-AP6-mediated depolarization can be inhibited by the non-NMDA receptor antagonist CNQX in ODAP- or QUIS-sensitized slices, the NMDA antagonist D-AP5 inhibits depolarization by cystine or L-AP6 in HCA-sensitized slices. Thus, HCA is the first identified NMDA agonist that induces phosphonate or cystine sensitization. Like QUIS sensitization, the sensitization evoked by either ODAP or HCA can be reversed by a subsequent exposure to 2 mM alpha-aminoadipic acid. Finally, we have demonstrated that there is a correlation between the potency of inducers for triggering phosphonate or cystine sensitivity and their affinities for System x(c)(-) and either the non-NMDA or NMDA receptor. Thus, the results of this study support our previous model of QUIS sensitization and have important implications for the mechanisms of neurotoxicity, neurolathyrism and hyperhomocystinemia.

Topics: 2-Aminoadipic Acid; 6-Cyano-7-nitroquinoxaline-2,3-dione; Amino Acids, Diamino; Animals; Cell Death; Cystine; Dose-Response Relationship, Drug; Electrophysiology; Excitatory Amino Acid Agonists; Excitatory Amino Acid Antagonists; Hippocampus; Homocysteine; In Vitro Techniques; Lathyrus; Male; Norleucine; Organophosphonates; Pyramidal Cells; Quisqualic Acid; Rats; Rats, Sprague-Dawley; Receptors, Glutamate; Receptors, N-Methyl-D-Aspartate; Receptors, Presynaptic

Glial cells synthesise neuroactive substances and release them upon neurotransmitter receptor activation. Homocysteic acid (HCA), an endogenous agonist for glutamatergic N-methyl-D-aspartate (NMDA) receptors, is predominantly localised in glial cells. We have previously demonstrated the release of HCA from mouse astrocytes in culture following activation of beta-adrenergic receptors. Moreover, a release of HCA has also been observed in vivo upon physiological stimulation of sensory afferents in the thalamus. Here we report the glutamate-induced release of HCA from astrocytes. The effect of glutamate was mediated by the activation of ionotropic (NMDA and non-NMDA) as well as by metabotropic receptors. In addition, the release of HCA was Ca(2+)- and Na(+)-dependent, and its mechanism involved the activation of the Na+/Ca(2+)-exchanger. Furthermore, we provide evidence for the presence of functional NMDA receptors on astrocytes, which are coupled to an intracellular Ca2+ increase via stimulation of the Na+/Ca(2+)-exchanger. Our data thus favour a participation of glial cells in excitatory neurotransmission and corroborate the role of HCA as a "gliotransmitter."

Topics: 6-Cyano-7-nitroquinoxaline-2,3-dione; Amiloride; Animals; Animals, Newborn; Astrocytes; Calcium; Cells, Cultured; Cerebral Cortex; Chromatography, High Pressure Liquid; Drug Interactions; Excitatory Amino Acid Agonists; Excitatory Amino Acid Antagonists; Glutamic Acid; Homocysteine; Mice; Models, Neurological; Neurons; Sulfur Isotopes

The sulphur-containing amino acid homocysteic acid (HCA) is present in and released in vitro from nervous tissue and is a potent neuronal excitant, predominantly activating N-methyl-d-aspartate (NMDA) receptors. However, HCA is localised not in neurones but in glial cells [Eur J Neurosci 3 (1991) 1370], and we have shown that it is released from astrocytes in culture upon glutamate receptor activation [Neuroscience 124 (2004) 377]. We now report the in vivo release of HCA from ventrobasal (VB) thalamus following natural stimulation of somatosensory afferents arising from the facial vibrissae of the rat. Simultaneously with multi-unit recording, [35S]-methionine, a HCA precursor, was perfused through a push-pull cannula in VB thalamus of anaesthetized rats. Perfusates were collected before, during and after 4 min stimulation of the vibrissal afferents with an air jet. A marked release of radiolabeled HCA was observed during and after the stimulation. Furthermore, the beta-adrenoreceptor agonist isoproterenol, which is known to evoke HCA release from glia in vitro, was found to increase the efflux of HCA in the perfusate in vivo. In separate experiments, the excitatory actions of iontophoretically applied HCA on VB neurones were inhibited by the NMDA receptor antagonist CPP, but not by the non-NMDA antagonist CNQX. These results suggest a possible "gliotransmitter" role for HCA in VB thalamus. The release of HCA from glia might exert a direct response or modulate responses to other neurotransmitters in postsynaptic neurons, thus enhancing excitatory processes.

Topics: 6-Cyano-7-nitroquinoxaline-2,3-dione; Action Potentials; Animals; Brain Chemistry; Chromatography, High Pressure Liquid; Excitatory Amino Acid Agonists; Excitatory Amino Acid Antagonists; Homocysteine; Iontophoresis; Kainic Acid; Male; Methionine; N-Methylaspartate; Neuroglia; Physical Stimulation; Piperazines; Rats; Rats, Wistar; Sulfur Isotopes; Synaptic Transmission; Thalamus; Vibrissae

Depolarization-induced suppression of inhibition (DSI) is a process whereby brief approximately 1-s depolarization to the postsynaptic membrane of hippocampal CA1 pyramidal cells results in a transient suppression of GABA(A)ergic synaptic transmission. DSI is triggered by a postsynaptic rise in [Ca(2+)](in) and yet is expressed presynaptically, which implies that a retrograde signal is involved. Recent evidence based on synthetic metabotropic glutamate receptor (mGluR) agonists and antagonists suggested that group I mGluRs take part in the expression of DSI and raised the possibility that glutamate or a glutamate-like substance is the retrograde messenger in hippocampal CA1. This hypothesis was tested, and it was found that the endogenous amino acids L-glutamate (L-Glu) and L-cysteine sulfinic acid (L-CSA) suppressed GABA(A)-receptor-mediated inhibitory postsynaptic currents (IPSCs) and occluded DSI, whereas L-homocysteic acid (L-HCA) and L-homocysteine sulfinic acid (L-HCSA) did not. Activation of metabotropic kainate receptors with kainic acid (KA) reduced IPSCs; however, DSI was not occluded. When iontophoretically applied, both L-Glu and L-CSA produced a transient IPSC suppression similar in magnitude and time course to that observed during DSI. Both DSI and the actions of the amino acids were antagonized by (S)-alpha-methyl-4-carboxyphenylglycine ([S]-MCPG), indicating that the effects of the endogenous agonists were produced through activation of mGluRs. Blocking excitatory amino acid transport significantly increased DSI and the suppression produced by L-Glu or L-CSA without affecting the time constant of recovery from the suppression. Similar to DSI, IPSC suppression by L-Glu or L-CSA was blocked by N-ethylmaleimide (NEM). Moreover, paired-pulse depression (PPD), which is unaltered during DSI, is also not significantly affected by the amino acids. Taken together, these results support the glutamate hypothesis of DSI and argue that L-Glu or L-CSA are potential retrograde messengers in CA1.

Topics: 6-Cyano-7-nitroquinoxaline-2,3-dione; Animals; Cysteine; Evoked Potentials; Excitatory Amino Acids; Glutamic Acid; Hippocampus; Homocysteine; In Vitro Techniques; Male; Neurotransmitter Agents; Pyramidal Cells; Rats; Rats, Sprague-Dawley; Receptors, GABA-A; Synaptic Transmission

Depolarizations induced by a range of amino acids including some sulphur-containing excitatory transmitter candidates were evoked from motoneurones in the neonatal rat spinal cord under conditions that precluded activation of known ionotropic glutamate receptors. The responses could be partially and differentially depressed by continuous application of several metabotropic glutamate receptor (mGluR) antagonists or by receptor desensitization with the mGluR agonist, (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid [(1S,3R)-ACPD]. In most cases [the exceptions being (1S,3R)-ACPD and to a lesser extent, quisqualate], the major component of these depolarizations was resistant to antagonism by phenylglycine-derived mGluR antagonists or desensitization of (1S,3R)-ACPD-sensitive receptors. Of the excitatory responses observed with the tested agonists, those evoked by L-glutamate itself were generally the least affected by blockade of known glutamate receptors.

Topics: 6-Cyano-7-nitroquinoxaline-2,3-dione; Animals; Animals, Newborn; Excitatory Amino Acid Agonists; Excitatory Amino Acid Antagonists; Excitatory Amino Acids; Homocysteine; Motor Neurons; Quisqualic Acid; Rats; Receptors, Metabotropic Glutamate; Spinal Cord

When rabbit retinas are exposed in vitro to specific excitatory amino acid receptor agonists certain GABAergic amacrine cells are activated to cause a release of GABA. The GABA that is not released can be detected by immunohistochemistry. Exposure of tissues to kainate or NMDA each caused a characteristic change in the GABA immunoreactivity. CNQX antagonised the kainate effect specifically while MK-801 counteracted the influence of NMDA. The effect produced by kainate was mimicked by domoic acid while the influence of homocysteic acid was identical with NMDA. Flupirtine alone did not influence the nature of the GABA immunoreactivity and so did not act as a kainate or NMDA agonist. However, flupirtine counteracted the influence produced by NMDA and homocysteic acid but had no effect on the kainate and domoic acid responses. Thus in this system flupirtine acts as an NMDA antagonist.

Topics: 6-Cyano-7-nitroquinoxaline-2,3-dione; Aminopyridines; Animals; Dizocilpine Maleate; gamma-Aminobutyric Acid; Homocysteine; Immunohistochemistry; Kainic Acid; Rabbits; Receptors, N-Methyl-D-Aspartate; Retina

1. We have examined the effects of iontophoretic application of antagonists to excitatory amino acid (EAA) receptors, as well as glycine and gamma-aminobutyric acid (GABA), on rhythmically active (RA) brain stem neurons during cortically induced masticatory activity (RMA) in the anesthetized guinea pig. Ten of these neurons were antidromically activated at latencies of 0.3-0.9 ms by stimulation of the trigeminal motor nucleus (MoV). 2. RA neurons were divided into closer and opener type according to the phase of activation during RMA. Iontophoretic application of 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), a specific non-N-methyl-D-aspartate (NMDA) receptor antagonist, suppressed discharge of both closer and opener type RA neurons during RMA. In contrast, iontophoretic application of 3-((1)-2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid (CPP), a specific NMDA receptor antagonist, was not effective in suppressing discharge of most opener type RA neurons but did reduce activity of closer type RA neurons. 3. Spike discharge of most RA neurons was time locked to each cortical stimulus during RMA. Some of the RA neurons were activated at a short latency to short pulse train stimulation of the cortex in the absence of RMA. In most cases CNQX reduced such time-locked responses during RMA and greatly reduced discharge evoked by short pulse stimulation of the cortex in all cases. In contrast, CPP was not as effective in suppressing either the time-locked responses during RMA or the discharge evoked by short pulse train stimulation of the cortex. 4. D,L-Homocysteic acid (HCA) application produced low level maintained discharge in RA neurons before RMA induction. When RMA was evoked in combination with HCA, rhythmical burst discharges with distinct interburst periods during the opening phase of RMA were observed in most closer type RA neurons. In contrast, during RMA in combination with HCA application, opener type RA neurons showed burst discharges that were modulated during the RMA cycle but lacked distinct interburst periods during the closer phase of the cycle. 5. During application of strychnine (STR), a glycine antagonist, discharge of closer type RA neurons increased in the opener phase of RMA during continuous HCA application. In contrast, bicuculline methiodide (BIC), a GABA antagonist, did not increase unit discharge of closer type RA neurons in the opener phase of RMA. 6. It is concluded that closer type RA neurons receive, alternatively, EAA-mediated

Topics: 6-Cyano-7-nitroquinoxaline-2,3-dione; Animals; Brain Mapping; Brain Stem; Cerebral Cortex; Electric Stimulation; Excitatory Amino Acid Antagonists; GABA Antagonists; Guinea Pigs; Homocysteine; Interneurons; Mastication; Membrane Potentials; Neural Inhibition; Neural Pathways; Piperazines; Reaction Time; Receptors, Glycine; Receptors, N-Methyl-D-Aspartate; Strychnine; Synaptic Transmission; Trigeminal Nerve; Trigeminal Nuclei

1. Although the gill and siphon withdrawal reflex of Aplysia has been used as a model system to study learning-associated changes in synaptic transmission, the identity of the neurotransmitter released by the sensory neurons and excitatory interneurons of the network mediating this behavior is still unknown. The identification of the putative neurotransmitter of these neurons should facilitate further studies of synaptic plasticity in Aplysia. 2. We report that sensory-motor transmission within this circuit is mediated through the activation of an excitatory amino acid receptor that is blocked by the non-N-methyl-D-aspartate excitatory amino acid receptor antagonists 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and 1-(4-chlorobenzoyl)-piperazine-2,3-dicarboxylic acid (CBPD). Compound postsynaptic potentials evoked in motor neurons by electrical stimulation of the siphon nerve were blocked by 92% with CNQX (75 microM) and 89% with CBPD (75 microM). 3. Simultaneous intracellular recordings were obtained from sensory neurons, excitatory interneurons, and motor neurons. Monosynaptic excitatory postsynaptic potentials (EPSPs) evoked in motor neurons by an action potential in a sensory neuron were blocked by 86% with CNQX (75 microM) and 71% with CBPD (75 microM). The two antagonists also blocked monosynaptic interneuronal EPSPs onto motor neurons by 65% and 67%, respectively. 4. Potential agonists of the synaptic receptors were puff-applied in the intact abdominal ganglion. Homocysteic acid (HCA) was found to mimic the action of the synaptically released transmitter because it strongly excites motor neurons. This effect was blocked by CNQX. Kainate and domoic acid were also effective agonists. 5. The actions of L- and D-glutamate as well as quisqualate were found to be mainly hyperpolarizing, whereas aspartate and (+/-)-amino-3-hydroxy-5-methylisoxazole-4-propionic acid had no effect. 6. Several reasons may be proposed to explain the inability of puff-applied glutamate to excite effectively the postsynaptic neurons in the intact ganglion. It is possible nonetheless that other endogenous amino acids such as HCA act as neurotransmitters at these synapses.

Topics: 6-Cyano-7-nitroquinoxaline-2,3-dione; Amino Acids; Animals; Anticonvulsants; Aplysia; Calcium Channels; Central Nervous System; Ganglia, Invertebrate; Homocysteine; Interneurons; Motor Activity; Piperazines; Quinoxalines; Receptors, Amino Acid; Receptors, N-Methyl-D-Aspartate; Sensory Receptor Cells; Synapses; Synaptic Transmission

We have compared the kinetic properties of NMDA receptor channels activated by exogenous agonists with those activated synaptically. Short (4 msec) applications of L-glutamate to outside-out patches from hippocampal neurons evoked currents that decayed with a double exponential time course that was controlled by both the unbinding rate of agonist and receptor desensitization. Lower-affinity agonists evoked NMDA receptor-activated currents that had faster rates of decay and recovered from desensitization more quickly, consistent with the idea that agonists which dissociate faster allow the receptor to reach a desensitized state less often. Both synaptic and patch responses could be well fitted with a simple kinetic model comprised of two independent but identical binding sites, one open state, one closed state, and one desensitized state, all doubly liganded. Provided that the agonist has a slow unbinding rate relative to the rates into the open and desensitized states (e.g., L-glutamate), this model predicts a response with two decay phases and can thus account for the synaptic response. Since the unbinding rate is the critical determinant of the time course, different affinity transmitters would affect such properties as excitatory postsynaptic current (EPSC) duration. Of the known endogenous excitatory amino acids, only L-glutamate has an affinity for the NMDA receptor consistent with the time course of the EPSC recorded between hippocampal neurons in culture.

Topics: 6-Cyano-7-nitroquinoxaline-2,3-dione; Animals; Animals, Newborn; Aspartic Acid; Cells, Cultured; Cysteic Acid; Evoked Potentials; Glutamates; Hippocampus; Homocysteine; Kinetics; Neurons; Quinoxalines; Rats; Receptors, N-Methyl-D-Aspartate; Synapses; Time Factors

1. We have examined the actions and pharmacology of two putative optic nerve transmitters, N-acetylaspartylglutamate (NAAG) and L-homocysteic acid (L-HCA), in the feline dorsal lateral geniculate nucleus (dLGN). We compared the responses obtained to iontophoretic application of these substances with those elicited by visual stimulation and application of specific N-methyl-D-aspartate (NMDA) and non-NMDA receptor agonists. The relative effects of the selective NMDA antagonist 3-[(+/-)-2-carboxypiperazin-4-yl]-propyl-1-phosphonic acid (CPP) and the selective non-NMDA antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) were tested on these responses. 2. There was a pronounced contrast between the influence of iontophoretically applied NAAG and L-HCA on dLGN cells. Iontophoretic application of NAAG [ejection current range 75-200 nA (mean 125 nA)] evoked either no effect (17/37), or very weak and sluggish excitatory (16/37) or inhibitory (4/37) effects. Conversely, L-HCA application [current range 25-136 nA (mean 67 nA)] elicited brisk and powerful excitatory responses (32/32) that were comparable with those produced by visual stimulation and iontophoresis of NMDA, kainate, and alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA). 3. Responses to L-HCA were selectively antagonized by application of the NMDA receptor antagonist CPP but were generally much less affected by the non-NMDA receptor antagonist CNQX. The weak and inconsistent responses to NAAG were not compatible with an evaluation of antagonist effects. 4. CPP application at dose levels selective for NMDA with respect to kainate and AMPA did not exert equal effects on L-HCA and NMDA. Whereas the mean responses to L-HCA were reduced to 32% of control for Y cells and 21% for X cells, those to NMDA were 11 and 11%, respectively. However, the level of reduction of the visual response for X and Y cells was very similar to that of L-HCA, visual responses being reduced to 35 and 22% of control for Y and X cells. 5. CNQX application reduced the visual response level of Y cells to 64% of control and that of X cells to 65%. The mean level for the L-HCA response of Y cells was 106% of control; the mean for X cells, 79%, was substantially below control. The responses to kainate and AMPA were reduced to a much greater extent. 6. The data suggest that it is unlikely that NAAG is the optic nerve transmitter.(ABSTRACT TRUNCATED AT 400 WORDS)

Topics: 6-Cyano-7-nitroquinoxaline-2,3-dione; Animals; Cats; Dipeptides; Female; Geniculate Bodies; Homocysteine; Immunohistochemistry; Iontophoresis; Neurons; Neurotransmitter Agents; Photic Stimulation; Piperazines; Quinoxalines

1. A grease-gap technique has been used to measure d.c. potentials, in response to the application of excitatory amino acids and electrical stimulation of the Schaffer collateral-commissural pathway, in the CA1 region of rat hippocampal slices. The actions of L-glutamate (L-Glu) have been quantified and compared to those of structurally related compounds. 2. Perfusion of L-Glu (90s applications) depolarized the tissue with a threshold of approximately 50 microM and a maximum response in excess of 10 mM. L-Aspartate (L-Asp) produced a similar dose-response relationship. By comparison N-methyl-D-aspartate (NMDA) and alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) were more potent excitants, producing dose-dependent depolarizations over the range 2-50 microM. 3. Application of the agonists depressed the amplitude of electrically-evoked synaptic responses; an effect that presumably reflects depolarization of neuronal tissue. However, for a given agonist-induced d.c. potential. L-Glu or L-Asp caused smaller depressions of synaptic responses than did either NMDA or AMPA. 4. The combined application of 50 microM D-2-amino-5-phosphonopentanoate (AP5) and 10 microM 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) substantially depressed synaptic responses and antagonized responses to NMDA and AMPA producing mean (+/- s.e.) dose-ratios of 12.2 +/- 1.2 and 7.0 +/- 0.8, respectively. However, these compounds produced minimal antagonism of responses to L-Glu and L-Asp (dose-ratios of 1.5 +/- 0.1 and 1.5 +/- 0.2, respectively). 5. Responses to the stereoisomers of homocysteate (HCA) were compared over the range 50 microM to 10 mM. D-HCA was approximately 3.6 times more potent than L-HCA and was antagonized to a greater extent by the combined application of 50 microM AP5 and 10 microM CNQX; the dose ratios were 8.7 + 0.8 and 5.1 + 0.9 for the D- and L- isomers, respectively. 6. The application of high doses of an excitant (e.g., 50mM L-Glu or 5mM D-HCA) caused an irreversible loss of sensitivity to NMDA and AMPA and abolished synaptic transmission. Responses to the other excitants were depressed by this excitotoxic lesion in the following order: D-HCA > L-HCA > L-Glu = LAsp. In slices treated in this manner, L-Glu, L-Asp and L-HCA produced very similar dose-response curves. 7. Some slices were unresponsive to NMDA, AMPA and electrical stimulation from the onset of the experiment but had sensitivity to L-Glu, L-Asp and L-HCA similar to that of slices that had rece

Topics: 2-Amino-5-phosphonovalerate; 6-Cyano-7-nitroquinoxaline-2,3-dione; Animals; Biological Transport, Active; Electrophysiology; Glutamates; Glutamic Acid; Hippocampus; Homocysteine; In Vitro Techniques; N-Methylaspartate; Quinoxalines; Quisqualic Acid; Rats; Synaptic Transmission

beta-p-Chlorophenylglutamate (Chlorpheg), a specific L-homocysteate (L-HC) uptake blocker, was tested on the L-HC- and L-glutamate-induced currents and on the excitatory postsynaptic potentials (EPSPs) evoked in CA1 rat hippocampal neurons by Schaffer collaterals stimulation. In the presence of tetrodotoxin (TTX; 1 microM), Chlorpheg (0.5-2 mM) potentiated L-HC- but not L-glutamate-induced currents. In normal magnesium containing medium and at resting membrane potential, Chlorpheg (1.5-1 mM) increased the amplitude and duration of the EPSPs evoked by Schaffer collaterals stimulation. This effect was prevented by bath application of the N-methyl-D-aspartate (NMDA) receptor antagonist CPP (20 microM). Chlorpheg enhanced also the NMDA component of the EPSP, evoked in the presence of 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 10 microM), bicuculline (20 microM) and glycine (100 microM). This effect was blocked by CPP (20 microM). It is concluded that L-HC is an endogenous NMDA agonist at the Schaffer collateral-CA1 synapse.

Topics: 6-Cyano-7-nitroquinoxaline-2,3-dione; Animals; Bicuculline; Evoked Potentials; Glutamates; Glycine; Hippocampus; Homocysteine; In Vitro Techniques; Male; Piperazines; Quinoxalines; Rats; Rats, Inbred Strains; Receptors, N-Methyl-D-Aspartate; Synapses; Tetrodotoxin

Responses of neocortical pyramidal cells to excitatory amino acids were recorded intracellularly. Agonists and antagonists were applied electrophoretically from a separate multibarrel pipette and care taken to ensure that the pipette was positioned to evoke optimal responses to N-methyl-D-aspartate (NMDA), or homocysteic acid, before control responses were recorded. Responses to NMDA, but not those to alpha-amino-3-hydroxy-5-methyl-4-isoxazdepropionic acid (AMPA) or quisqualate, were enhanced when glycine was co-applied. Responses to AMPA, quisqualate and NMDA were reduced by 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) applied either electrophoretically, or in the bathing medium, with responses to quisqualate being the least and those to AMPA being the most sensitive to CNQX. The blockade of NMDA responses by CNQX was selectively reversed by additional glycine confirming that CNQX blocks NMDA receptor-channel complexes at the glycine, rather than at the NMDA site. Under control conditions, responses to glutamate resembled responses to quisqualate, and were relatively insensitive to CNQX, 3-((+/-)-2-carboxypiperazin-4-yl)-propyl-l-phosphonic acid and 2-amino-5-phosphonovalerate, while responses to homocysteic acid resembled responses to NMDA and were blocked by these antagonists. This suggested that homocysteic acid acted at NMDA receptors, while glutamate acted primarily at non-NMDA receptors. However, responses to both glutamate and homocysteic acid were augmented by additional glycine when these transmitter candidates were applied close to a "hot spot" for NMDA receptor activation. The glycine enhancement of responses to glutamate was sensitive to NMDA antagonists, indicating that glutamate can activate NMDA receptors in an intact preparation if glycine levels are high enough.

Topics: 6-Cyano-7-nitroquinoxaline-2,3-dione; Action Potentials; alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid; Amino Acids; Animals; Cerebral Cortex; Drug Interactions; Electrophysiology; Female; Glycine; Homocysteine; Ibotenic Acid; In Vitro Techniques; Kinetics; Male; Membrane Potentials; N-Methylaspartate; Neurons; Quinoxalines; Quisqualic Acid; Rats; Somatosensory Cortex