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0270-647419151999Aug01The Journal of neuroscience : the official journal of the Society for NeuroscienceJ NeurosciAssociation of AMPA receptors with a subset of glutamate receptor-interacting protein in vivo.652865376528-37The NMDA and AMPA classes of ionotropic glutamate receptors are concentrated at postsynaptic sites in excitatory synapses. NMDA receptors interact via their NR2 subunits with PSD-95/SAP90 family proteins, whereas AMPA receptors bind via their GluR2/3 subunits to glutamate receptor-interacting protein (GRIP), AMPA receptor-binding protein (ABP), and protein interacting with C kinase 1 (PICK1). We report here a novel cDNA (termed ABP-L/GRIP2) that is virtually identical to ABP except for additional GRIP-like sequences at the N-terminal and C-terminal ends. Like GRIP (which we now term GRIP1), ABP-L/GRIP2 contains a seventh PDZ domain at its C terminus. Using antibodies that recognize both these proteins, we examined the subcellular localization of GRIP1 and ABP-L/GRIP2 (collectively termed GRIP) and their biochemical association with AMPA receptors. Immunogold electron microscopy revealed the presence of GRIP at excitatory synapses and also at nonsynaptic membranes and within intracellular compartments. The association of native GRIP and AMPA receptors was confirmed biochemically by coimmunoprecipitation from rat brain extracts. A majority of detergent-extractable GluR2/3 was complexed with GRIP in the brain. However, only approximately half of GRIP was associated with AMPA receptors. Unexpectedly, immunocytochemistry of cultured hippocampal neurons and rat brain at the light microscopic level showed enrichment of GRIP in GABAergic neurons and in GABAergic nerve terminals. Thus GRIP is associated with inhibitory as well as excitatory synapses. Collectively, these findings support a role for GRIP in the synaptic anchoring of AMPA receptors but also suggest that GRIP has additional functions unrelated to the binding of AMPA receptors.WyszynskiMMDepartment of Neurobiology and Howard Hughes Medical Institute, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts 02114, USA.ValtschanoffJ GJGNaisbittSSDunahA WAWKimEEStandaertD GDGWeinbergRRShengMMengNS35050NSNINDS NIH HHSUnited StatesR01 NS034361NSNINDS NIH HHSUnited StatesCA66268CANCI NIH HHSUnited StatesR01 NS035050NSNINDS NIH HHSUnited StatesR29 NS034361NSNINDS NIH HHSUnited StatesNS35527NSNINDS NIH HHSUnited StatesF32 CA066268CANCI NIH HHSUnited StatesR01 NS035527NSNINDS NIH HHSUnited StatesJournal ArticleResearch Support, Non-U.S. Gov'tResearch Support, U.S. Gov't, P.H.S.
United StatesJ Neurosci81021400270-64740Carrier Proteins0DNA, Recombinant0Grip1 protein, rat0Grip2 protein, rat0Intercellular Signaling Peptides and Proteins0Intracellular Signaling Peptides and Proteins0Nerve Tissue Proteins0Receptors, AMPAIMAmino Acid SequencegeneticsAnimalsBrainmetabolismultrastructureCarrier ProteinsgeneticsmetabolismCells, CulturedDNA, RecombinantHippocampuscytologymetabolismIntercellular Signaling Peptides and ProteinsIntracellular Signaling Peptides and ProteinsNerve Tissue ProteinsgeneticsmetabolismNeuronsmetabolismRatsRats, Sprague-DawleyReceptors, AMPAmetabolismSequence Homology, Amino AcidTissue Distributionphysiology1999722199972201199972200200021ppublish10414981PMC678283010.1523/JNEUROSCI.19-15-06528.1999Allison DW, Gelfand VI, Spector I, Craig AM. Role of actin in anchoring postsynaptic receptors in cultured hippocampal neurons: differential attachment of NMDA versus AMPA receptors. J Neurosci. 1998;18:2423–2436.PMC67930949502803Banker GA, Cowan WM. 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