title: Rabphilin 3A retains NMDA receptors at synaptic sites through interaction with GluN2A/PSD-95 complex.
abstract: NMDA receptor (NMDAR) composition and synaptic retention represent pivotal features in the physiology and pathology of excitatory synapses. Here, we identify Rabphilin 3A (Rph3A) as a new GluN2A subunit-binding partner. Rph3A is known as a synaptic vesicle-associated protein involved in the regulation of exo- and endocytosis processes at presynaptic sites. We find that Rph3A is enriched at dendritic spines. Protein-protein interaction assays reveals that Rph3A N-terminal domain interacts with GluN2A(1349-1389) as well as with PSD-95(PDZ3) domains, creating a ternary complex. Rph3A silencing in neurons reduces the surface localization of synaptic GluN2A and NMDAR currents. Moreover, perturbing GluN2A/Rph3A interaction with interfering peptides in organotypic slices or in vivo induces a decrease of the amplitude of NMDAR-mediated currents and GluN2A density at dendritic spines. In conclusion, Rph3A interacts with GluN2A and PSD-95 forming a complex that regulates NMDARs stabilization at postsynaptic membranes.
authors.affiliation: DiSFeB, Dipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, Via Balzaretti 9, Milano 20133, Italy.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Stanic
authors.affiliation: Institut Interdisciplinaire de Neurosciences, University of Bordeaux, CNRS UMR 5297, Bordeaux 33000, France.
authors.forename: Mario
authors.initials: M
authors.lastname: Carta
authors.affiliation: DiSFeB, Dipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, Via Balzaretti 9, Milano 20133, Italy.
authors.forename: Ivano
authors.initials: I
authors.lastname: Eberini
authors.affiliation: DiSFeB, Dipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, Via Balzaretti 9, Milano 20133, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Pelucchi
authors.affiliation: DiSFeB, Dipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, Via Balzaretti 9, Milano 20133, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Marcello
authors.affiliation: Dipartimento di Scienze del Farmaco, Università degli Studi del Piemonte Orientale 'Amedeo Avogadro', Novara 28100, Italy.
authors.forename: Armando A
authors.initials: AA
authors.lastname: Genazzani
authors.affiliation: Institute of Neuroscience, Newcastle University, Newcastle upon Tyne NE2 4HH, UK.
authors.forename: Claudia
authors.initials: C
authors.lastname: Racca
authors.affiliation: Institut Interdisciplinaire de Neurosciences, University of Bordeaux, CNRS UMR 5297, Bordeaux 33000, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Mulle
authors.affiliation: DiSFeB, Dipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, Via Balzaretti 9, Milano 20133, Italy.
authors.forename: Monica
authors.initials: M
authors.lastname: Di Luca
authors.affiliation: DiSFeB, Dipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, Via Balzaretti 9, Milano 20133, Italy.
authors.forename: Fabrizio
authors.initials: F
authors.lastname: Gardoni
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D000818:Animals; D056547:CA1 Region, Hippocampal / Q000378:metabolism*; D019556:COS Cells; D002522:Chlorocebus aethiops; D003198:Computer Simulation; D049229:Dendritic Spines / Q000378:metabolism*; D000076122:Disks Large Homolog 4 Protein; D004705:Endocytosis; D051528:Guanylate Kinases / Q000378:metabolism*; D006624:Hippocampus / Q000378:metabolism; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D018613:Microscopy, Confocal; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism*; D054731:PDZ Domains; D018408:Patch-Clamp Techniques; D051381:Rats; D016194:Receptors, N-Methyl-D-Aspartate / Q000378:metabolism*; D013569:Synapses / Q000378:metabolism; D013570:Synaptic Membranes / Q000378:metabolism*; D033921:Vesicular Transport Proteins / Q000378:metabolism*; D000097624:Rabphilin-3A
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D000076122:Disks Large Homolog 4 Protein; D051528:Guanylate Kinases; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D016194:Receptors, N-Methyl-D-Aspartate; D033921:Vesicular Transport Proteins; D000097624:Rabphilin-3A; C501447:Dlg4 protein, mouse; C501449:Dlg4 protein, rat