Early onset of NMDA receptor GluRε1 (NR2A) expression and its abundant postsynaptic localization in developing motoneurons of the mouse hypoglossal nucleus

Oshima, Shohei; Fukaya, Masahiro; Nagashima, Masato; Shirakawa, Tetsuo; Oguchi, Haruhisa; Watanabe, Masahiko

doi:10.1016/s0168-0102(02)00035-4

Cited by 26 publications

(30 citation statements)

References 59 publications

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“…Here we show definitive ultrastructural evidence of this switch in hippocampal synapses. This appears to be the common pattern in the central nervous system (Erisir and Harris, 2003;Liu et al, 2004) although there are exceptions, such as for some motoneurons that express abundant NR2A early in development (Oshima et al, 2002). In Sans et al (2000), we also showed the switch in the MAGUK composition of hippocampus using both immunoblots and immunogold localization at synapses (i.e., SAP102 decreases and PSD-95 and PSD-93 increase).…”

Section: Discussionsupporting

confidence: 55%

Ontogeny of postsynaptic density proteins at glutamatergic synapses

Petralia

Sans

Wang

et al. 2005

Molecular and Cellular Neuroscience

209

196

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In glutamatergic synapses, glutamate receptors (GluRs) associate with many other proteins involved in scaffolding and signal transduction. The ontogeny of these postsynaptic density (PSD) proteins involves changes in their composition during development, paralleling changes in GluR type and function. In the CA1 region of the hippocampus, at postnatal day 2 (P2), many synapses already have a distinct PSD. We used immunoblot analysis, subcellular fractionation, and quantitative immunogold electron microscopy to examine the distribution of PSD proteins during development of the hippocampus. Synapses at P2 contained substantial levels of NR1 and NR2B and most GluRassociated proteins, including SAP102, SynGAP, the chain of proteins from GluRs/SAP102 through GKAP/Shank/Homer and metabotropic glutamate receptors, and the adhesion factors, cadherin, catenin, neuroligin, and Nr-CAM. Development was marked by substantial decreases in NR2B and SAP102 and increases in NR2A, PSD-95, AMPA receptors and CaMKII. Other components showed more moderate changes.

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Section: Discussionsupporting

confidence: 55%

Ontogeny of postsynaptic density proteins at glutamatergic synapses

Petralia

Sans

Wang

et al. 2005

Molecular and Cellular Neuroscience

209

196

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“…A, C, E, and G: taken using ϫ10 objective, scale bar ϭ 250 m. B, D, F, and H: taken using ϫ60 objective, scale bar ϭ 25 m. Monyer et al 1994). In the mouse hypoglossal motor nucleus, there is a high expression of NMDAR ⑀1 subunit (mouse equivalent of NR2A) at a very early age, appearing at embryonic day 13 (Oshima et al 2002). Our results show NFPS blocks glycine uptake sufficiently to result in potentiation of the NMDA response in both age groups.…”

Section: Discussionmentioning

confidence: 61%

Blockade of Glycine Transporter-1 (GLYT-1) Potentiates NMDA Receptor-Mediated Synaptic Transmission in Hypoglossal Motorneurons

Lim

Hoang

Berger

2004

Journal of Neurophysiology

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of glycine transporter-1 (GLYT-1) potentiates NMDA receptor-mediated synaptic transmission in hypoglossal motorneurons. J Neurophysiol 92: 2530 -2537, 2004. First published June 2, 2004 10.1152/ jn.01123.2003. NMDA receptor (NMDAR)-mediated spontaneous miniature excitatory postsynaptic currents (mEPSCs) are potentiated by exogenously applied glycine. In this study, we have investigated the effect of blocking glycine uptake on NMDAR-mediated responses from hypoglossal motorneurons (HMs) of rats. We have used N[3-(NFPS; 500 nM), an antagonist of glycine transporter-1 (GLYT1), to study the effect of blocking endogenous glycine uptake on NMDAR-mediated synaptic transmission. We show that the charge transfer of NMDARmediated mEPSCs was enhanced after NFPS application in neonate (P2-4) and juvenile rats (P8-11), but this enhancement was statistically significant only in the former group. Spontaneous and evoked EPSCs showed a significant increase in NMDAR-mediated charge transfer in both neonates and juveniles. The greater increase observed in spontaneous EPSCs may be due to increased release of glycine from glycinergic terminals in the absence of tetrodotoxin (TTX). Brief application of NMDA onto HMs showed that extrasynaptic NMDARs may be potentiated by NFPS only in the presence of extracellularly applied glycine. Immunohistochemistry of GLYT1 and -2 shows labeling throughout the hypoglossal nucleus. GLYT1 labeling is diffuse and becomes more intense and uniform during development consistent with its glial localization. In contrast, GLYT2 labeling is intense throughout the nucleus and increases in intensity with age. Our results demonstrate the glycine binding site of the NMDAR is not saturated in the brain stem slice during the first 2 wk of development. We suggest that modulation of glycine concentration by GLYT1 is an important mechanism to regulate NMDAR-mediated synaptic transmission.

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“…In immunohistochemistry for NMDA and AMPA receptor subunits and PSD-95/synapse-associated protein 90 (SAP90), paraffin sections (5 m) of control and mutant mice were subjected to pepsin pretreatment, as reported previously Fukaya and Watanabe, 2000;Yamada et al, 2001;Oshima et al, 2002). The sections were immunoreacted with rabbit or guinea pig primary antibodies (all at 1 g/ml) against GluN2AC (formerly termed GluR1C), GluN2BN (GluR2 N), GluN2BC (GluR2C) , GluN1-C2 (GluR1-C2) subunits of NMDARs (Abe et al, 2004), GluA1 (GluR1, GluR-A, or GluR␣1) subunit of AMPA receptors (AMPARs) , PSD-95 (Fukaya and Watanabe, 2000), ␤-galactosidase (ab616; Abcam), and Cre recombinase (69050-3; Novagen).…”

Section: Methodsmentioning

confidence: 99%

NMDA Receptor GluN2B (GluRε2/NR2B) Subunit Is Crucial for Channel Function, Postsynaptic Macromolecular Organization, and Actin Cytoskeleton at Hippocampal CA3 Synapses

Akashi¹,

Kakizaki²,

Kamiya³

et al. 2009

J. Neurosci.

145

107

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GluN2B (GluR2/NR2B) subunit is involved in synapse development, synaptic plasticity, and cognitive function. However, its roles in synaptic expression and function of NMDA receptors (NMDARs) in the brain remain mostly unknown because of the neonatal lethality of global knock-out mice. To address this, we generated conditional knock-out mice, in which GluN2B was ablated exclusively in hippocampal CA3 pyramidal cells. By immunohistochemistry, GluN2B disappeared and GluN1 (GluR1/NR1) was moderately reduced, whereas GluN2A (GluR1/NR2A) and postsynaptic density-95 (PSD-95) were unaltered in the mutant CA3. This was consistent with protein contents in the CA3 crude fraction: 9.6% of control level for GluN2B, 47.7% for GluN1, 90.6% for GluN2A, and 98.0% for PSD-95. Despite the remaining NMDARs, NMDAR-mediated currents and long-term potentiation were virtually lost at various CA3 synapses. Then, we compared synaptic NMDARs by postembedding immunogold electron microscopy and immunoblot using the PSD fraction. In the mutant CA3, GluN1 was severely reduced in both immunogold (20.6-23.6%) and immunoblot (24.6%), whereas GluN2A and PSD-95 were unchanged in immunogold but markedly reduced in the PSD fraction (51.4 and 36.5%, respectively), indicating increased detergent solubility of PSD molecules. No such increased solubility was observed for GluN2B in the CA3 of GluN2A-knock-out mice. Furthermore, significant decreases were found in the ratio of filamentous to globular actin (49.5%) and in the density of dendritic spines (76.2%). These findings suggest that GluN2B is critically involved in NMDAR channel function, organization of postsynaptic macromolecular complexes, formation or maintenance of dendritic spines, and regulation of the actin cytoskeleton.

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Early onset of NMDA receptor GluRε1 (NR2A) expression and its abundant postsynaptic localization in developing motoneurons of the mouse hypoglossal nucleus

Cited by 26 publications

References 59 publications

Ontogeny of postsynaptic density proteins at glutamatergic synapses

Ontogeny of postsynaptic density proteins at glutamatergic synapses

Blockade of Glycine Transporter-1 (GLYT-1) Potentiates NMDA Receptor-Mediated Synaptic Transmission in Hypoglossal Motorneurons

NMDA Receptor GluN2B (GluRε2/NR2B) Subunit Is Crucial for Channel Function, Postsynaptic Macromolecular Organization, and Actin Cytoskeleton at Hippocampal CA3 Synapses

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