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PMID: 19726645 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

NMDA receptor GluN2B (GluR epsilon 2/NR2B) subunit is crucial for channel function, postsynaptic macromolecular organization, and actin cytoskeleton at hippocampal CA3 synapses.

The Journal of neuroscience : the official journal of the Society for Neuroscience ·Vol. 29 ·No. 35 ·2009-09-02 ·Pages 10869-82

Akashi K, Kakizaki T, Kamiya H, Fukaya M, Yamasaki M, Abe M, Natsume R, Watanabe M, Sakimura K

Abstract

GluN2B (GluRepsilon2/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 (GluRzeta1/NR1) was moderately reduced, whereas GluN2A (GluRepsilon1/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.

MeSH Terms
Actins/genetics,physiology Animals Cells, Cultured Cytoskeleton/genetics,physiology Dendrites/physiology Excitatory Postsynaptic Potentials/genetics Gene Knock-In Techniques Hippocampus/chemistry,physiology Macromolecular Substances/chemistry Mice Mice, Inbred C57BL Mice, Knockout Mice, Transgenic Protein Subunits/deficiency,genetics,physiology Receptors, N-Methyl-D-Aspartate/deficiency,genetics,physiology Synapses/genetics,physiology
Chemicals
Actins Macromolecular Substances NR2B NMDA receptor Protein Subunits Receptors, N-Methyl-D-Aspartate
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Akashi Kaori
Department of Cellular Neurobiology, Brain Research Institute, Niigata University, Niigata 951-8585, Japan.
Kakizaki Toshikazu
Kamiya Haruyuki
Fukaya Masahiro
Yamasaki Miwako
Abe Manabu
Natsume Rie
Watanabe Masahiko
Sakimura Kenji
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Article Info
Journal
The Journal of neuroscience : the official journal of the Society for Neuroscience
Abbr.
J Neurosci
ISSN
1529-2401
DOI
10.1523/JNEUROSCI.5531-08.2009
Published
2009-09-02
Pages
10869-82
Language
English
Region
United States
NLM ID
8102140
PMCID
PMC6665524
Subset
IM
External Links
PubMed source DOI: 10.1523/JNEUROSCI.5531-08.2009 PMC Full Text View journal details
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