Molecular cloning and characterization of the rat NMDA receptor

Moriyoshi, Koki; Masu, Masayuki; Ishii, Takahiro; Shigemoto, Ryuichi; Mizuno, Noboru; Nakanishi, Shigetada

doi:10.1038/354031a0

Cited by 1,702 publications

(923 citation statements)

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“…However, the mechanisms by which these effects of estrogens are yet to be fully characterized. N-methyl-D-aspartate (NMDA) receptors, a subtype of ionotropic glutamate receptor, have been implicated, as mediators, in some effects of estradiol on morphological plasticity and related physiological and cognitive processes in the brain (Moriyoshi et al 1991). However, only few studies have directly investigated the role of estradiol in regulating NMDA receptors in rat and, in addition, these studies have focused on the effects of estradiol on hippocampal NMDA receptors (Woolley and McEwen 1992;Gazzaley et al 1996;Woolley et al 1997).…”

Section: Hormonal Specificity Of Modulation Of N-methyl-d-aspartate (mentioning

confidence: 99%

“…The measurements of the total area of eight adjacent sections of the pyramidal cell layer of the hippocampal CA1 and CA2/3 regions and the molecular layer dentate gyrus (Bregma Ϫ4.20 mm) (Paxinos and Watson 1982) were performed to evaluate NMDA receptor subunits 1, 2A, and 2B mRNA levels. In situ hybridization to evaluate NMDA receptors subunits 2A and 2B mRNA levels in cortical and striatal regions of the brain was not performed because of the lower level of expression found in these regions (Moriyoshi et al 1991, Monyer et al 1992. Examples are shown in Figure 2.…”

Section: In Situ Hybridization Of Nmda Receptorsmentioning

confidence: 99%

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Estrogen-like Activity of Tamoxifen and Raloxifene on NMDA Receptor Binding and Expression of its Subunits in Rat Brain

Cyr

Thibault

Morissette

et al. 2001

Neuropsychopharmacology

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Hormonal specificity of modulation of N-methyl-D-aspartate (NMDA) receptors was investigated by comparing the effects of estradiol with tamoxifen or raloxifene, which display different responses in breast, bone, and uterus. Two weeks ovariectomy in rats decreasedIt has been shown that the female sex steroid estradiol exerts profound effects on neuronal differentiation during development (Toran-Allerand et al. 1983) and recently, accumulating evidence support also a modulatory role of estrogens in the normal maintenance of brain function during aging (Simpkins et al. 1994). Beneficial effects of estradiol in mood disorders (i.e., depression, pre-menstrual syndrome, post-natal depression), as well as mental diseases, such as schizophrenia and Gille de la Tourette's syndrome, have been reported (Di Paolo 1994;Fink et al. 1998). Moreover, a beneficial role of estradiol in neurodegenerative disFrom the Oncology and Molecular Endocrinology Research Center, Laval University Medical Center, (CHUL), and Faculty of Pharmacy, Laval University, Sainte-Foy, Quebec, Canada, G1V 4G2.Address correspondence to: Dr. Thérèse Di Paolo, Oncology and Molecular Endocrinology Research Center, Laval University Medical Center, CHUQ, 2705, Laurier Boulevard, Sainte-Foy, Quebec, Canada G1V 4G2.Received July 5, 2000; revised December 20, 2000; accepted January 3, 2001. N EUROPSYCHOPHARMACOLOGY 2001 -VOL . 25 , NO . 2 SERMs Modulate NMDA Receptors 243 eases, such as Alzheimer has been recently suggested (Tang et al. 1996;Henderson 1997;Inestrosa et al. 1998). However, the mechanisms by which these effects of estrogens are yet to be fully characterized. N-methyl-D-aspartate (NMDA) receptors, a subtype of ionotropic glutamate receptor, have been implicated, as mediators, in some effects of estradiol on morphological plasticity and related physiological and cognitive processes in the brain (Moriyoshi et al. 1991). However, only few studies have directly investigated the role of estradiol in regulating NMDA receptors in rat and, in addition, these studies have focused on the effects of estradiol on hippocampal NMDA receptors (Woolley and McEwen 1992;Gazzaley et al. 1996;Woolley et al. 1997). We have recently investigated the effects of estradiol on the NMDA receptors agonist sites in brain regions implicated in mental disorders, such as frontal cortex, striatum, and nucleus accumbens (Cyr et al. 2000a). Our experiments showed that ovariectomy, as well as estradiol have regional specific effects since, for example, ovariectomy decreases NMDA receptor specific binding in hippocampal CA1 and dentate gyrus regions, whereas it has no effect in frontal cortex. In contrast, estradiol increases NMDA specific binding in the hippocampal CA1 and the dentate gyrus, but it decreases this binding in the frontal cortex (Cyr et al. 2000a).The present study sought to determine whether changes observed previously at the protein level after hormonal withdrawal due to ovariectomy or after estradiol treatment reflect changes at the level of gene transcription...

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Section: Hormonal Specificity Of Modulation Of N-methyl-d-aspartate (mentioning

confidence: 99%

Section: In Situ Hybridization Of Nmda Receptorsmentioning

confidence: 99%

Estrogen-like Activity of Tamoxifen and Raloxifene on NMDA Receptor Binding and Expression of its Subunits in Rat Brain

Cyr

Thibault

Morissette

et al. 2001

Neuropsychopharmacology

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“…4 The receptors are hetero-oligomeric proteins formed by obligatory NR1 subunits 5,6 interacting with NR2A-D, conferring functional variability. 7,8 The major NR2 subunits in adult neocortex and hippocampus are NR2A and NR2B. Although they are not exclusive locations, 9 NR2A is predominantly confined to synapses of mature neurons 5 while NR2B is distributed mainly extrasynaptically.…”

Section: Introductionmentioning

confidence: 99%

Excitotoxicity and focal cerebral ischemia induce truncation of the NR2A and NR2B subunits of the NMDA receptor and cleavage of the scaffolding protein PSD-95

et al. 2007

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The N-methyl-D-aspartate receptor (NMDAR) is central to physiological and pathological functioning of neurons. Although promising results are beginning to be obtained in the treatment of dementias, clinical trials with NMDAR antagonists for stroke, trauma and neurodegenerative disorders, such as Hungtinton's disease, have failed before. In order to design effective therapies to prevent excitotoxic neuronal death, it is critical to characterize the consequences of excessive NMDAR activation on its expression and function. Previous data have reported partial downregulation of the NR1 and NR2B receptor subunits in response to excitotoxicity and cerebral ischemia. However, the effect of NMDAR overactivation on NR2A, a subunit fundamental to synaptic transmission and neuronal survival, is still elusive. In this study, we report the rapid and extensive proteolytic processing of NR2A, together with the scaffolding protein postsynaptic density-95 (PSD-95), induced by excitotoxic stimulation of cortical neurons in vitro and by transient focal cerebral ischemia. Processing of the C terminus of NR2A is irreversibly induced by brief agonist exposure of NR2B-containing receptors, and requires calcium influx and the activity of calpain, also responsible for PSD-95 cleavage. The outcome is a truncated NR2A subunit that is stable and capable to interact with NR1 at the surface of neurons, but lacking the structural domains required for association with scaffolding, downstream signaling and cytoskeletal proteins. Therefore, a rapid and significant uncoupling of synaptic NMDARs from downstream survival pathways is expected to occur during ischemia. This novel mechanism induced by excitotoxicity helps to explain the failure of most therapies based on NMDAR antagonists.

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“…For the construction of pNR1-dM4CR, PCRs were performed with primer F (5h-GTATCCTAGGCATGGTGTGGG-3h) and each of primer R1 (5h-GGATCCTCATCAGTGATGATGA-TGATGATGCTCAAAAGTGAGGGTTGCAGGAG-3h) and primer R2 (5h-GGATCCTCATCATAGATCCTCCTCTGAG-ATGAGCTTTTGCTCCTCAAAAGTGAGGGTTGCAG-GAG-3h), with pN60 [5] as a template. BlnI-BamHI fragments (1.0 kbp) cut out of the PCR products and a 1.9 kbp SacI-BlnI fragment of pN60 were ligated into SacI\BamHI-digested pUC118, resulting in pUC118NR1his and pUC118NR1myc respectively.…”

Section: Construction Of the Transfer Vectors And The Recombinant Bacmentioning

confidence: 99%

“…This property made it possible to clone the NR1 cDNA by the oocyte expression system [5]. In succession, four NR2 subunits (NR2A-D) were identified [6][7][8].…”

Section: Introductionmentioning

confidence: 99%

Expression and characterization of a glycine-binding fragment of the N-methyl-D-aspartate receptor subunit NR1

Miyazaki¹,

Nakanishi

Jingami³

1999

Biochemical Journal

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N-Methyl--aspartate receptor channels are composed of an NR1 subunit and at least one of the NR2 subunits (NR2A-D). Activation of the N-methyl--aspartate receptor requires the coagonists glycine and glutamate. It has been proposed that the NR1 subunit possesses a glycine-binding site. We have expressed a soluble form of the NR1 subunit, which was produced by connecting the N-terminal extracellular region with the extracellular loop between the third and fourth membrane segments, by a baculovirus system along with full-length and truncated membrane-bound forms. The soluble NR1 receptor was efficiently secreted into the culture medium and showed a high affinity for ligands. The K d of a glycine-site antagonist, [$H]MDL 105,519[(E)-3-(2-phenyl-2-carboxyethenyl)-4,6-dichloro-1H-indole-2-carboxylic acid], for the soluble receptor was 3.89p0.97 nM, which was comparable to the K d of 4.47p1.39 nM for the membrane-bound full-length form. These values were close to the values reported previously with the use

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Molecular cloning and characterization of the rat NMDA receptor

Cited by 1,702 publications

References 43 publications

Estrogen-like Activity of Tamoxifen and Raloxifene on NMDA Receptor Binding and Expression of its Subunits in Rat Brain

Estrogen-like Activity of Tamoxifen and Raloxifene on NMDA Receptor Binding and Expression of its Subunits in Rat Brain

Excitotoxicity and focal cerebral ischemia induce truncation of the NR2A and NR2B subunits of the NMDA receptor and cleavage of the scaffolding protein PSD-95

Expression and characterization of a glycine-binding fragment of the N-methyl-D-aspartate receptor subunit NR1

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