Abstract
N-methyl-D-aspartate (NMDA) receptors (NMDARs) play a central role in development, synaptic plasticity, and neurological disease. NMDAR subunit composition defines their biophysical properties and downstream signaling. Casein kinase 2 (CK2) phosphorylates the NR2B subunit within its PDZ-binding domain; however, the consequences for NMDAR localization and function are unclear. Here we show that CK2 phosphorylation of NR2B regulates synaptic NR2B and NR2A in response to activity. We find that CK2 phosphorylates NR2B, but not NR2A, to drive NR2B-endocytosis and remove NR2B from synapses resulting in an increase in synaptic NR2A expression. During development there is an activity-dependent switch from NR2B to NR2A at cortical synapses. We observe an increase in CK2 expression and NR2B phosphorylation over this same critical period and show that the acute activity-dependent switch in NR2 subunit composition at developing hippocampal synapses requires CK2 activity. Thus, CK2 plays a central role in determining the NR2 subunit content of synaptic NMDARs.
Copyright © 2010 Elsevier Inc. All rights reserved.
Publication types
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Research Support, N.I.H., Intramural
MeSH terms
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Adenosine Triphosphate / pharmacokinetics
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Amino Acid Sequence
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Animals
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Animals, Newborn
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Benzimidazoles / pharmacology
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Biotinylation / methods
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Casein Kinase II / physiology*
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Cells, Cultured
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Cerebral Cortex / cytology
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Disks Large Homolog 4 Protein
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Embryo, Mammalian
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Endocytosis / genetics
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Excitatory Amino Acid Antagonists / pharmacology
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Excitatory Postsynaptic Potentials / drug effects
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Excitatory Postsynaptic Potentials / genetics
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Gene Expression Regulation / genetics
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Green Fluorescent Proteins / genetics
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Hippocampus / cytology
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Hippocampus / metabolism
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Humans
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Immunoprecipitation / methods
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In Vitro Techniques
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Intracellular Signaling Peptides and Proteins / metabolism
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Membrane Proteins / metabolism
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Models, Biological
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Neurons / drug effects
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Neurons / metabolism*
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PDZ Domains / physiology
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Patch-Clamp Techniques
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Phosphorus Isotopes / pharmacokinetics
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Phosphorylation / drug effects
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Phosphorylation / genetics
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Piperidines / pharmacology
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Protein Subunits / genetics
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Protein Subunits / metabolism*
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Rats
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Rats, Sprague-Dawley
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Rats, Wistar
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Receptors, N-Methyl-D-Aspartate / genetics
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Receptors, N-Methyl-D-Aspartate / metabolism*
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Sodium Channel Blockers / pharmacology
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Subcellular Fractions / drug effects
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Subcellular Fractions / metabolism
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Synapses / metabolism*
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Synaptophysin / metabolism
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Tetrodotoxin / pharmacology
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Transfection / methods
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Tyrosine / metabolism
Substances
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4,5,6,7-tetrabromobenzimidazole
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Benzimidazoles
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Disks Large Homolog 4 Protein
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Dlg4 protein, rat
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Excitatory Amino Acid Antagonists
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Intracellular Signaling Peptides and Proteins
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Membrane Proteins
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NR2A NMDA receptor
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Phosphorus Isotopes
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Piperidines
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Protein Subunits
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Receptors, N-Methyl-D-Aspartate
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Sodium Channel Blockers
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Synaptophysin
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Green Fluorescent Proteins
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Tyrosine
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Tetrodotoxin
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Adenosine Triphosphate
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Casein Kinase II
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ifenprodil