Identification and Characterization of an <i>N</i>‐Methyl‐D‐Aspartate‐Specific L‐[<sup>3</sup>H]Glutamate Recognition Site in Synaptic Plasma Membranes

Monahan, Joseph B.; Michel, Jeffrey B.

doi:10.1111/j.1471-4159.1987.tb05726.x

Cited by 132 publications

(53 citation statements)

References 45 publications

(38 reference statements)

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“…(Slevin & Kasarskis, 1985). However, zinc has relatively little action on the specific binding of L-glutamate to NMDA receptors, since with 1 mM-zinc ZINC MODULATION OF GLUTAMATE RECEPTORS receptor occupancy was reduced by only 24% (Monahan & Michel, 1987 (Reynolds & Miller, 1988), similar to the value of 11 ,UM estimated in the present experiments. These results suggest that at micromolar concentrations zinc does not act as a competitive antagonist, nor interfere greatly with the binding of agonists to the NMDA receptor.…”

Section: Perfusion Techniquessupporting

confidence: 76%

Modulation of excitatory amino acid receptors by group IIB metal cations in cultured mouse hippocampal neurones.

Mayer

Vyklicky

Westbrook

1989

The Journal of Physiology

255

143

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SUMMARY1. Responses to the excitatory amino acids kainate, quisqualate, N-methyl-Daspartate (NMDA), L-glutamate and L-aspartate were recorded in mouse hippocampal neurones in cell culture, using the whole-cell configuration of the patch clamp technique. Agonists were applied rapidly from an array of flow pipes each of 250 ,um diameter, positioned within 100 ,um of the nerve cell body.2 control, while responses to kainate and quisqualate were increased to 1-08 and 1 15 times control. With 1 mM-cadmium responses to NMDA were reduced to 0 04 times control while responses to kainate and quisqualate were reduced to 0 79 and 0-60 times control. Mercury was neurotoxic and increased the leakage current; however, no reduction of the response to NMDA was produced by 5/M-mercury. 4. The equilibrium dissociation constant (Kd) for zinc antagonism of responses to NMDA, estimated from fit of a single binding site adsorption isotherm, was 13 ,UM; cadmium was about 4 times less potent than zinc. These effects of zinc and cadmium were nearly voltage independent. In contrast the antagonism of responses to NMDA by 150 /tm-magnesium was highly voltage dependent, such that the Kd for magnesium increased e-fold per 17-6 mV depolarization.

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Section: Perfusion Techniquessupporting

confidence: 76%

Modulation of excitatory amino acid receptors by group IIB metal cations in cultured mouse hippocampal neurones.

Mayer

Vyklicky

Westbrook

1989

The Journal of Physiology

255

143

View full text Add to dashboard Cite

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“…In addition, Bmax values of these glutamate subtypes in human brains were about 1/20 of those found in rat brain, although the Kd values were equivalent (5). Moreover, 100 ,uM KA inhibited [3H]glutamate binding by 50% in this experiment and by 18 -30% in the previous reports using the rat brain (11,12). These findings suggest a species difference.…”

Section: Discussionsupporting

confidence: 37%

Reassessment of (3H)glutamate binding to human brain membrane preparations.

et al. 1991

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ABSTRACT-We studied [3H]glutamate binding to human brain membrane preparations. We found that the binding assay of [3H]glutamate should be done at 4°C using glass-fiber filters pretreated with polyethyleneimine, sterile incubation buffer, and membrane preparations pretreated with a detergent. Under these assay conditions, we have characterized the three subtypes of glutamate receptors: N-methyl-D-aspartate-, quisqualate-and kainate-sensitive binding sites. Scatchard analysis revealed that the N-methyl-D-aspartate-sensitive [3H]glutamate binding sites consisted of a single component with a dissociation constant (Kd) of 28.2 ± 2.8 nM and total concentration (Bmax) of 36.2 ± 3.6 fmol/mg protein (N = 8). These data were much different from those presented in previous studies.

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“…Monahan & Michel, 1987) and the strychnine-insensitive glycine binding site. This latter is separate from the aspartate/NMDA site and potentiates the NMDA response (Johnson & Ascher, 1987); the presence of glycine has been suggested to be required for NMDA receptor activation (Kleckner & Dingledine.…”

Section: Analysis Of Single-channel Currents In Cell-attached and Insmentioning

confidence: 99%

Pharmacological properties and H+ sensitivity of excitatory amino acid receptor channels in rat cerebellar granule neurones.

Traynelis

Cull-Candy

1991

The Journal of Physiology

196

106

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SUMMARY1. N-Methyl-D-aspartate (NMDA), a-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA), and kainate receptor channels have been exainined in rat cerebellar granule neurones with whole-cell and single-channel patch-clamp methods. The whole-cell peak and steady-state aspartate and NMDA currents were reversibly inhibited by extracellular protons; the IC50 (concentration producing half-maximal inhibition) for the full H+ inhibition curve for NMDA receptors corresponde,d to pH 7 3, near to physiological pH. (S)-AMPA and kainate whole-cell currents were inhibited by protons with IC50 values that corresponded to pH 6-3 and 57, respectively; these receptors were, however, insensitive to H+ concentrations that inhibited NMDA receptor responses.2. Proton inhibition of the NMDA, AMPA and kainate receptor-mediated responses was voltage insensitive, and did not involve a shift in reversal potential.3. The EC50 (concentration producing half-maximal effect) for aspartate calculated from the whole-cell dose-response curve was similar at pH 6-8 and 7-6 (mean 11-2 ,SM). Although the EC50 for glycine potentiation of the aspartate response was marginally increased from 273 nM at pH 7-6 to 373 nm at pH 648, H+ inhibition was not overcome by up to 1 mM-external glycine. Inhibiting concentrations of H+ appropriate for AMPA and kainate receptors did not markedly alter the EC50 values determined for (S)-AMPA (3-4 gM) and kainate (114 #M) at pH 7-2.4. Treatment of neurones with N-ethylmaleimide, iodoacetic acid, dithiothretiol or diethyl pyrocarbonate did not influence proton inhibition of NMDA receptor responses. However, treatment with diethyl pyrocarbonate, which potentiated aspartate responses, appeared to reduce the effectiveness of Zn2+ inhibition of NMDA receptors.5. Desensitization of whole-cell NMDA and (S)-AMPA currents was studied with ionophoretic application of agonist to the cell soma. Whole-cell aspartate currents desensitized rapidly, irrespective of the glycine concentration. Increased H+ concentrations did not detectably alter the ratio of peak/steady-state current, or the time constants describing the onset of, or recovery from, desensitization. The time constant describing desensitization of (S)-AMPA-induced whole-cell currents also appeared unchanged by inhibiting pH (6 2). MS 8533 S. F. TRAYNELIS AND S. G. CULL-CANVDY6. The amplitudes of aspartate-or NMDA-activated single-channel multiple conductance levels were unchanged by decreasing the pH to 6-8. Similarly, concentrations of H+ that inhibit (S)-AMPA (pH 6 2) and kainate currents (pH 5 6) did not change the single-channel conductance for either respective channel, as estimated from fluctuation analysis of whole-cell currents.7. Recording of NMDA receptor channels from outside-out, inside-out, and cellattached patches suggested that protons act at a site on or near the extracellular face of the receptor.8. The opening frequency both of aspartate-activated individual openings and bursts of openings was decreased by 58-2 and 49 5 % with reductions in pH fro...

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Identification and Characterization of an N‐Methyl‐D‐Aspartate‐Specific L‐[³H]Glutamate Recognition Site in Synaptic Plasma Membranes

Cited by 132 publications

References 45 publications

Modulation of excitatory amino acid receptors by group IIB metal cations in cultured mouse hippocampal neurones.

Modulation of excitatory amino acid receptors by group IIB metal cations in cultured mouse hippocampal neurones.

Reassessment of (3H)glutamate binding to human brain membrane preparations.

Pharmacological properties and H+ sensitivity of excitatory amino acid receptor channels in rat cerebellar granule neurones.

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Identification and Characterization of an N‐Methyl‐D‐Aspartate‐Specific L‐[3H]Glutamate Recognition Site in Synaptic Plasma Membranes

Cited by 132 publications

References 45 publications

Modulation of excitatory amino acid receptors by group IIB metal cations in cultured mouse hippocampal neurones.

Modulation of excitatory amino acid receptors by group IIB metal cations in cultured mouse hippocampal neurones.

Reassessment of (3H)glutamate binding to human brain membrane preparations.

Pharmacological properties and H+ sensitivity of excitatory amino acid receptor channels in rat cerebellar granule neurones.

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Identification and Characterization of an N‐Methyl‐D‐Aspartate‐Specific L‐[³H]Glutamate Recognition Site in Synaptic Plasma Membranes