non-NMDA Type Glutamate Receptors Research Articles

Neurosteroid Actions in Memory and Neurologic/Neuropsychiatric Disorders.

Memory dysfunction is a symptomatic feature of many neurologic and neuropsychiatric disorders; however, the basic underlying mechanisms of memory and altered states of circuitry function associated with disorders of memory remain a vast unexplored territory. The initial discovery of endogenous neurosteroids triggered a quest to elucidate their role as neuromodulators in normal and diseased brain function. In this review, based on the perspective of our own research, the advances leading to the discovery of positive and negative neurosteroid allosteric modulators of GABA type-A (GABAA), NMDA, and non-NMDA type glutamate receptors are brought together in a historical and conceptual framework. We extend the analysis toward a state-of-the art view of how neurosteroid modulation of neural circuitry function may affect memory and memory deficits. By aggregating the results from multiple laboratories using both animal models for disease and human clinical research on neuropsychiatric and age-related neurodegenerative disorders, elements of a circuitry level view begins to emerge. Lastly, the effects of both endogenously active and exogenously administered neurosteroids on neural networks across the life span of women and men point to a possible underlying pharmacological connectome by which these neuromodulators might act to modulate memory across diverse altered states of mind.

Enantioselective Synthesis of (-)-Dysiherbaine.

Dysiherbaine, a natural product isolated from the Marine sponge Dysidea herbacea, has been shown to be a selective agonist of non-NMDA type glutamate receptors, kainate receptors. An enantioselective synthesis of dysiherbaine is reported. Metathesis of the diene followed by conversion of the resulting alkene to the amino alcohol and addition of the amino acid provides the natural product. This synthesis differs from previous approaches to the molecule in that the functionality on the tetrahydropyran ring is installed late in the route.

Fast Inhibition of Glutamate-Activated Currents by Caffeine

BackgroundCaffeine stimulates calcium-induced calcium release (CICR) in many cell types. In neurons, caffeine stimulates CICR presynaptically and thus modulates neurotransmitter release.Methodology/Principal FindingsUsing the whole-cell patch-clamp technique we found that caffeine (20 mM) reversibly increased the frequency and decreased the amplitude of miniature excitatory postsynaptic currents (mEPSCs) in neocortical neurons. The increase in mEPSC frequency is consistent with a presynaptic mechanism. Caffeine also reduced exogenously applied glutamate-activated currents, confirming a separate postsynaptic action. This inhibition developed in tens of milliseconds, consistent with block of channel currents. Caffeine (20 mM) did not reduce currents activated by exogenous NMDA, indicating that caffeine block is specific to non-NMDA type glutamate receptors.Conclusions/SignificanceCaffeine-induced inhibition of mEPSC amplitude occurs through postsynaptic block of non-NMDA type ionotropic glutamate receptors. Caffeine thus has both pre and postsynaptic sites of action at excitatory synapses.

The role of substance P in the glutamatergic transmission in the reticular formation near the nucleus ambiguus in rats

In order to estimate the neuronal mechanism of switching autonomic reactions in medulla oblongata, we examined the response of neurons in the reticular formation near the nucleus ambiguus (NA) to the administration of substance P (SP). Whole-cell recording was applied to the neurons near the NA in coronal slice preparations of medulla oblongata isolated from infant rat. In the 8 of 18 neurons, bath application of SP (1μM) increased the frequency of spontaneous activities. In contrast, 4 neurons showed inhibited response by application of SP, and other 6 neurons showed no response. Several neurons were clamped at −60mV and recorded EPSCs evoked by electrical stimulation of adjacent area to the recording neuron. Bath application of SP induced excitatory effects on the both amplitude of evoked inward EPSCs and frequency of spontaneous activities. These effects of SP were abolished by application of non-NMDA type glutamate receptor antagonists, CNQX. Our previous studies have shown that administration of neurokinin-1 receptor (one of the a representative receptor of SP) antagonist near the NA inhibited gastric and respiratory movement in anesthetized rat. These results suggested that SP acts as neuromediator in the glutamatergic transmission near NA and participated in generating certain autonomic reactions.

Mechanism of acute ischemic injury of oligodendroglia in early myelinating white matter: the importance of astrocyte injury and glutamate release.

In developing CNS white matter (WM), the period of early myelination is characterized by a heightened sensitivity to ischemic injury. Using an in situ (isolated) preparation, we show that the mechanism of acute ischemic injury of immature WM oligodendroglial involves Ca2+ influx though non-NMDA type glutamate receptors (GluRs). The Ca2+-influx and acute cell death that was evoked by ischemic conditions (oxygen and glucose withdrawal) in identified P10 rat optic nerve oligodendroglia were blocked by removing extracellular Ca2+ or by CNQX, a selective non-NMDA GluR antagonist. The selective Na-K-Cl cotransport (NKCC) inhibitor bumetanide was also highly protective, even though NKCC expression is restricted to astrocytes in this tissue. Bumetanide largely prevented the non-NMDA GluR-mediated [Ca2+]i rise evoked by ischemia in oligodendroglia, suggesting that it interfered with ischemic glutamate release. In control WM, glutamate-like reactivity was located mainly in astrocytes and oligodendroglia identified using ultrastructural criteria. In ischemic WM, astrocyte glutamate-like reactivity was reduced, an effect countered by bumetanide. We suggest a model in which NKCC-dependent injury and release of glutamate from astrocytes activates glutamate receptors on oligodendroglia, resulting in Ca2+-influx and acute cell death.

Glutamate-induced cobalt uptake reveals non-NMDA receptors in developing rat taste buds.

Non-NMDA type glutamate receptors are present in rat taste buds. However, the function of those receptors is not yet known. Developmental changes in the glutamate receptors in taste cells may provide clues to their functional role. We used a cobalt staining technique to determine at which stage in development functional non-NMDA glutamate receptors first appeared. Cobalt-stained taste bud cells first appeared in 20-day-old rats. The number of cobalt-stained cells increased with age and reached a maximum at 45 days. The shape of stained cells looked similar at all age groups. Cobalt-labeled cells appeared to be correlated with synaptic, not taste, glutamate receptors.

Analysis of spontaneous EPSCs in retinal horizontal cells of the carp.

Spontaneous excitatory postsynaptic currents (sEPSCs) were recorded under Whole-cell voltage clamp from carp type 1 horizontal cells (H1 cells) uncoupled by dopamine in retinal slices. Red light steps, which hyperpolarise cones and reduce glutamate release, induced outward current responses accompanied by a suppression of sEPSCs. sEPSCs decayed exponentially with a mean time constant of 0.71+/-0.07 ms and had a reversal potential near 0 mV. Power spectral analysis of sEPSCs revealed a similar decay time constant. They were suppressed by a non-NMDA receptor antagonist, CNQX at 10 microM, and a relatively specific AMPA receptor antagonist, GYKI52466 at 20 microM. The presence of sEPSCs suggests that the release of glutamate from cone synaptic terminals is vesicular. The reduction in mean sEPSC frequency with red light was not accompanied by a significant change in the mean sEPSC conductance increase (482+/-59 pS), suggesting that a decrease in the vesicular release rate from cones does not alter the vesicular glutamate concentration (quantal contents). The results suggest that the spontaneous events in H1 cells were contributed by non-NMDA (possibly AMPA) type glutamate receptors modulated by the red cone input.

Inhibitory effects of theanine and sera from theanine-fed rats on receptor-mediated cancer cell invasion beneath mesothelial-cell monolayers.

To investigate the bioavailability and mode of action of theanine against cancer, we examined in vitro and ex vivo effects of theanine on invasion of a rat ascites hepatoma cell line of AH109A. Theanine dose-dependently inhibited the invasion of AH109A cells across rat mesentery-derived mesothelial-cell (M-cell) monolayers without restraining AH109A cell proliferation in vitro. Rat sera obtained after oral intubation of theanine also inhibited the invasion. A competitive N-methyl-D-aspartate (NMDA) type glutamate receptor antagonist, (+/-) 2-amino-5-phosphonopentanoic acid (AP-5), dose-dependently counteracted the theanine-mediated in vitro and ex vivo inhibition of AH109A invasion. A competitive non-NMDA type glutamate receptor antagonist, 6,7-dinitroquinoxaline 2,3-dione (DNQX), did not affect this inhibition by theanine in vitro. These results suggest that the inhibition of AH109A invasion by theanine may be mediated by the NMDA receptor of AH109A.

GABA B receptor-mediated presynaptic inhibition of glutamatergic and GABAergic transmission in the basolateral amygdala

The information processing at central synapses is mediated not only by homosynaptic transmission with direct synaptic connections but also by heterosynaptic interactions between distinct synaptic inputs. Using rat brain slices and whole-cell recordings this study aimed to examine the roles of GABA B receptors in synaptic interactions in the basolateral amygdala (BLA), a critical brain structure related to fear and anxiety. Stimulation in the BLA produced non-NMDA type glutamate receptor antagonist-sensitive excitatory postsynaptic currents (EPSCs) and bicuculline-sensitive inhibitory postsynaptic currents (IPSCs) in the BLA neurons. The GABA B receptor agonist baclofen markedly inhibited both EPSCs and IPSCs in a concentration-dependent manner, and the baclofen-induced inhibition was selectively abolished by the GABA B receptor antagonist CGP55845A. The paired-pulse ratio of EPSC and IPSC amplitude was increased by baclofen. The effect of baclofen was mimicked by lowering the external Ca 2+ concentration but not by glutamate- and GABA A-receptor antagonists. The frequency but not the mean amplitude of miniature EPSCs and IPSCs was decreased by baclofen. The findings suggest that activation of GABA B receptors by baclofen reduces the strength of excitatory and inhibitory transmission in the BLA by a presynaptic mechanism. Repetitive conditioning stimulation applied to GABAergic synaptic inputs exerted an inhibitory action on glutamatergic excitatory transmission, and the stimulation-induced inhibition was abolished by CGP55845A. Furthermore, the paired-pulse ratio of EPSCs was increased during the stimulation-induced inhibition. The results in this study provide evidence that synaptic activation of GABA B heteroreceptors elicits presynaptic inhibition of glutamatergic excitatory transmission in the BLA.

Glucagon-like peptide-1 modulates neuronal activity in the rat's hippocampus.

In order to assess effects of glucagon-like peptide-1 (GLP-1) in the brain excluding the hypothalamus, the effects of GLP-1 (7-36) amide, a naturally produced active fragment, on the electroencephalogram and hippocampal single unit activities of anesthetized male Wistar rats were examined. I.c.v. injection of GLP-1 (7-36) amide decreased the hippocampal theta wave duration. Juxtacellular administration of GLP-1 (7-36) amide first increased and then decreased single unit activities recorded in the hippocampal CA1, which effects were antagonized by exendin (9-39) amide, a GLP-1 receptor antagonist, or 6-cyano-7-nitroquinoxaline-2,3-dione, a non-NMDA type glutamate receptor antagonist. These results indicate that GLP-1 receptors exist in the hippocampus and are involved in modulating hippocampal activity through an increase in the release of excitatory amino acid transmitters.

Dysiherbaine: A New Neurotoxic Amino Acid from the Micronesian Marine Sponge Dysidea herbacea

A new amino acid, dysiherbaine (1), was isolated from a Micronesian sponge Dysidea herbacea. The structure was determined by using FABMS, ESIMS, FABMS/CID/MS, and one- and two-dimensional NMR experiments of 1 and its dimethyl derivative 3 to be a novel diamino dicarboxylic acid, which consisted of a cis-fused hexahydrofuro[3,2-b]pyran ring substituted with a 3-[2-aminopropanoic acid] side chain. The relative configuration of the bicyclic portion of 1 was determined by 3JH,H analysis and difference NOE experiments, and that of the acyclic side chain was assigned by additional 2,3JC,H analysis, measured by hetero half-filtered TOCSY (HETLOC) and phase sensitive HMBC experiments. Systemic administration of 1 induced neurotoxic symptoms in mice which were reminiscent of neuroexcitatory amino acids such as domoic acid. Dysiherbaine inhibited bindings of [3H]-kainic acid (KA) and [3H]-1-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), but not [3H]CGS-19755, an N-methyl-d-asparatic acid (NMDA) receptor antagonist, on rat brain synaptic membranes, suggesting that 1 is a selective agonist of non-NMDA type glutamate receptors in the central nervous system.

Inhibition of spinal monosynaptic reflex in newborn rats by aurintricarboxylic acid

The effect of aurintricarboxylic acid (ATA), an inhibitor of nuclease, on glutamatergic synaptic transmission was examined electrophysiologically in the isolated spinal cords of newborn rats. Monosynaptic reflex (MSR) was depressed about 20%, 50 min after exposure to 100 μM of ATA. Pretreatment with APV, a N-methyl- d-aspartate (NMDA) type receptor antagonist, depressed MSR by about 10%, but additional application of ATA did not affect the MSR further. In contrast, the remaining MSR following treatment with DNQX, a non-NMDA type receptor antagonist, in the Mg 2+-free medium was almost completely inhibited by addition of ATA. ATA depressed NMDA- but not d, l- α-amino-3-hydroxy-5-methyl-4-isoxalone propionic acid (AMPA)- or kainate-induced depolarization in the medium containing normal ionic composition. Thus it is concluded that the reduction of MSR by ATA is due to blockade of NMDA type but non-NMDA type glutamate receptors. The present study also confirmed the previous conclusion that Ia monosynaptic transmission in the spinal cord of the newborn rat is mediated by NMDA as well as non-NMDA type glutamate receptors.

216 Production and characterization of mice devoid of the glutamic acid decarboxylase (67 kDa isoform-GAD67)

The information processing at central synapses is mediated not only by homosynaptic transmission with direct synaptic connections but also by heterosynaptic interactions between distinct synaptic inputs. Using rat brain slices and whole-cell recordings this study aimed to examine the roles of GABAB receptors in synaptic interactions in the basolateral amygdala (BLA), a critical brain structure related to fear and anxiety. Stimulation in the BLA produced non-NMDA type glutamate receptor antagonist-sensitive excitatory postsynaptic currents (EPSCs) and bicuculline-sensitive inhibitory postsynaptic currents (IPSCs) in the BLA neurons. The GABAB receptor agonist baclofen markedly inhibited both EPSCs and IPSCs in a concentration-dependent manner, and the baclofen-induced inhibition was selectively abolished by the GABAB receptor antagonist CGP55845A. The paired-pulse ratio of EPSC and IPSC amplitude was increased by baclofen. The effect of baclofen was mimicked by lowering the external Ca2+ concentration but not by glutamate- and GABAA-receptor antagonists. The frequency but not the mean amplitude of miniature EPSCs and IPSCs was decreased by baclofen. The findings suggest that activation of GABAB receptors by baclofen reduces the strength of excitatory and inhibitory transmission in the BLA by a presynaptic mechanism. Repetitive conditioning stimulation applied to GABAergic synaptic inputs exerted an inhibitory action on glutamatergic excitatory transmission, and the stimulation-induced inhibition was abolished by CGP55845A. Furthermore, the paired-pulse ratio of EPSCs was increased during the stimulation-induced inhibition. The results in this study provide evidence that synaptic activation of GABAB heteroreceptors elicits presynaptic inhibition of glutamatergic excitatory transmission in the BLA.

NMDA (NMDAR1) and AMPA-type (GluR2/3) receptor subunits are expressed in the inner ear.

Using receptor subunit-specific antibodies, the cellular localization of NMDA and AMPA type glutamate receptor subunits was studied within the rodent (rat, guinea pig) and non-human primate (monkey) inner ear. In the spiral and vestibular ganglion, almost all cells were immunoreactive for the NMDAR1 subunit and the AMPA type receptor subunit GluR2/3. This indicates that both NMDA and non-NMDA type glutamate receptors may be co-distributed in the primary afferent neuronal components, and are possibly involved in neurotransmission in the primary auditory and vestibular systems. This study also indicated the possible localizations of glutamate receptors in the nonneuronal cells in the inner ear, suggesting that some nonneuronal cells may also have the ability to mediate glutamate signalling.

Inhibition by suramin and reactive blue 2 of GABA and glutamate receptor channels in rat hippocampal neurons

Effects of suramin and reactive blue 2 (RB2), compounds known as antagonists at P2-purinoceptors, on ionic currents mediated through GABA and glutamate receptor channels were investigated in rat hippocampal neurons. Under whole-cell voltage-clamp, suramin (30 to 300 microM) and RB2 (10 to 100 microM) inhibited a current activated by 10 microM GABA in a concentration-dependent manner. Suramin (100 and 300 microM) and RB2 (10 and 30 microM) also inhibited an inward current activated by kainic acid (100 microM), an agonist at non-NMDA type glutamate receptor channels, and an inward current activated by N-methyl-D-aspartate (NMDA; 100 microM), an agonist at NMDA type glutamate receptor channels. The inhibition by suramin or RB2 did not exhibit voltage-dependence between -30 and -90 mV in the case of the GABA- or the kainate-evoked current. In contrast, the inhibition by these compounds of the NMDA-evoked current exhibited voltage-dependence and was enhanced by hyperpolarization. When the concentration of agonists was increased by 5- or 10-fold, the magnitude of the inhibition by suramin of the kainate-evoked current and the magnitude of the inhibition by RB2 of the NMDA-evoked current were attenuated. alpha,beta-Methylene ATP (100 microM) did not affect the GABA-, kainate- or NMDA-activated current. The results suggest that suramin and RB2 inhibit GABA receptor channels and glutamate receptor channels. The blockade of these channels must be taken into account when these compounds are used as pharmacological tools to examine an involvement of P2-purinoceptors, especially in preparations where GABAergic or glutamatergic neurotransmission is expected.

Regulation of brain-derived neurotrophic factor (BDNF) expression and release from hippocampal neurons is mediated by non-NMDA type glutamate receptors

We have examined the influence of glutamate on cortical brain-derived neurotrophic factor (BDNF) expression using in situ hybridization and immunohistochemistry. Kainic acid (KA) produced an upregulation of hippocampal and neocortical BDNF mRNA as well as BDNF protein that was blocked by a non-NMDA antagonist, 6,7-dinitroquinoxaline-2,3-dione (DNQX), but was not affected by the NMDA antagonist 2-amino-7-phosphonoheptanoic acid (AP7). Basal levels of BDNF mRNA were not affected by NMDA, DNQX, or AP7 treatment. BDNF protein was also increased after kainate exposure with a spatial and temporal course distinct from that seen for the expression of BDNF mRNA. A dramatic shift in BDNF immunoreactivity (-IR) was observed from intracellular compartments to the neuropil surrounding CA3 pyramidal cells 2-3 hr after KA exposure. This shift in localization of BDNF-IR suggests a constitutive release of BDNF at the level of the cell body and dendrites. Moreover, we have localized mRNAs for full-length and truncated trkB, to a co-incident population of neurons and glia. These data suggest the neurons that produce BDNF also express components necessary for a biological response to the same neurotrophic factor. The present study also demonstrates increased BDNF-IR in the mossy fiber terminal zone of hippocampus after exposure to KA, as well as an increase in trkB mRNA, and provides evidence of local release of this neurotrophin into the surrounding neuropil where it would be available for local utilization. The synthesis and putative release of BDNF from somatic and/or dendritic sites within the hippocampus provide evidence of a potential autocrine or paracrine role for BDNF, and establish a local source of trophic support for the maintenance of synaptic plasticity and anatomic reorganization in the mature nervous system.

Sympathetic and parasympathetic ganglia express non-NMDA type glutamate receptors: distinct receptor subunit composition in the principle and SIF cells

The presence of non-NMDA glutamate receptors in the rat sympathetic and parasympathetic ganglia was examined by immunocytochemistry using specific antibodies against AMPA-type excitatory amino acid receptor subunits (GluR1–4). Three kinds of antibodies specific to the GluR1, GluR2 and 3, and GluR4 subunits were used. The superior cervical ganglion and pterygopalatine ganglion were examined as representatives of sympathetic and parasympathetic ganglia. In the superior cervical ganglion, GluR1- and GluR2/3-like immunoreactivity was observed in most principal neurons and SIF cells. In contrast, GluR4-like immunoreactivity was not observed in the principal cells; however, SIF cells exhibited intense immunoreactivity of GluR4. In the pterygopalatine ganglion, the profile of the immunoreactivity was similar to that seen in the superior cervical ganglia. The subunit compositions between the principal cells and SIF cells were different, whereas the compositions in cell species involved in the autonomic ganglia, sympathetic and parasympathetic ganglia were identical. This suggests that glutamate is another important preganglionic transmitter together with acetylcholine, and the responses elicited in the principal cells and SIF cells might be different because of the difference in subunit composition.

Characteristic localization of non-NMDA type glutamate receptor subunits in the rat pituitary gland

While the involvement of the glutamate receptors in the hypothalamo-hypophyseal system has been clarified at the hypothalamic level, the existence of glutamate receptors in the pituitary gland has remained obscure. We investigated the localization of the glutamate receptors, the non-NMDA type receptor subunits (GluR1-4) in particular, by immunocytochemistry using specific antibodies. The antibodies specific to GluR1, GluR2/3 and GluR4 exhibited the characteristic localization of the receptor molecules in each lobe of the pituitary gland. GluR1- and GluR2/3-positive cells were identified in the anterior and intermediate lobe, and intense terminals of GluR4 and weak terminals of GluR2/3 were observed in the posterior lobe. Such immunoreactivity appeared to be at the axonal terminal of the neurosecretory magnocellular cells. This was confirmed by in situ hybridization histochemistry using specific oligodeoxynucleotide probes and by immunocytochemistry in the neurosecretory magnocellular neurons. The GluR4 mRNA-positive signal and GluR4 immunoreactivity were abundantly observed in magnocells of paraventricular and supraoptic nuclei. In addition to the positive fibers, some pituicytes in the posterior lobe exhibited GluR2/3 immunoreactivity. This suggests that pituicytes have non-NMDA type glutamate receptors. Thus, present study suggests that some anterior pituitary cells and pituicytes in the neural lobe are regulated by the glutamate.

Differential ontogenesis of presynaptic and postsynaptic GABAB inhibition in rat somatosensory cortex.

1. The postnatal maturation of gamma-aminobutyric acid (GABA)B receptor-mediated presynaptic inhibition was studied in brain slices of rat somatosensory cortex maintained in vitro. Patchclamp techniques were used to record whole-cell inhibitory post-synaptic currents from layer II-III neurons in animals from postnatal days (P) 7-24. Monosynaptic inhibitory postsynaptic currents (IPSCs) were evoked after N-methyl-D-aspartate (NMDA) and non-NMDA type glutamate receptors had been blocked by D-amino-phosphonovaleric acid (D-AP5, 20 microM) and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 microM), respectively. These IPSCs were solely mediated by postsynaptic GABAA receptors because they were abolished by bicuculline (10 microM), reversed polarity near the chloride equilibrium potential, and were recorded with electrodes that contained Cs+ to block postsynaptic GABAB responses. 2. When pairs of stimuli separated by intervals of 0.1-10 s were used to evoke IPSCs, the second response was depressed, an effect that was maximal at 300 ms. Evoked IPSCs were also depressed by baclofen (10 microM). The paired pulse depression (PPD) of monosynaptic IPSCs was decreased or eliminated by 2-OH-saclofen (200 microM). These findings indicate that PPD of monosynaptic IPSCs was due to presynaptic GABAB receptor-mediated inhibition of GABA release. 3. There were no significant differences in the amounts of PPD in neurons from different age groups (P7-10, P12-17, P22-24) at any interstimulus interval tested (0.1-10 s).(ABSTRACT TRUNCATED AT 250 WORDS)

The excitatory and inhibitory amino acid receptors on horizontal cells isolated from the white perch retina.

1. The distribution and the properties of receptors to the inhibitory amino acid glycine (GLY) and the excitatory amino acid glutamate (GLU) and its analogues kainate (KA), quisqualate (QUIS), alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), and N-methyl-D-aspartate (NMDA), were studied with whole-cell and outside-out patch-clamp techniques on all four types of horizontal cells isolated from the retina of white perch. 2. Glycine at concentrations above 30 microM evoked whole-cell current responses from two types of horizontal cells (H2 and H4). The other two types of horizontal cells (H1 and H3) were unresponsive to GLY (30 microM-3 mM). 3. Responses elicited by GLY from H2 and H4 cells were similar, consisting of inward currents that desensitized with a half-decay time of 0.5-2 s at glycine concentrations between 100 and 500 microM. GLY-activated currents were inhibited by the glycine receptor antagonist strychnine (STRYCH). Current responses evoked by GLY reversed at the Cl- equilibrium potential. 4. Dose-response analysis of peak currents induced by GLY revealed a Hill coefficient of 2.0 +/- 0.1 (mean +/- SD, n = 3) and an median effective concentration (EC50) of 85 +/- 2 microM (n = 3). 5. Single glycine receptor channels recorded from outside-out patches had a main-state conductance of 47 +/- 4 pS (n = 3). 6. Every type of horizontal cell from the white perch responded to GLU, KA, QUIS, and AMPA but none responded to exogenously applied NMDA (200 microM) or NMDA (200 microM) + GLY (1 microM) in a Mg+2-free bathing solution. 7. The ratio of the amplitude of responses to GLU, KA, QUIS, and AMPA remained nearly constant among all the horizontal cells tested, suggesting there might be only a single population of non-NMDA receptors on these cells. 8. QUIS and KA both elicited responses from the horizontal cells. When applied together with KA, QUIS competitively antagonized the responses of horizontal cells to KA. 9. The results demonstrated the existence of an inhomogeneous distribution of strychnine-sensitive glycine receptors and a homogeneous distribution of non-NMDA type glutamate receptors among the four types of white perch horizontal cells.