Rabphilin 3A retains NMDA receptors at synaptic sites through interaction with GluN2A/PSD-95 complex

Nat Commun. 2015 Dec 18;6:10181. doi: 10.1038/ncomms10181.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / metabolism*
  • Animals
  • CA1 Region, Hippocampal / metabolism*
  • COS Cells
  • Chlorocebus aethiops
  • Computer Simulation
  • Dendritic Spines / metabolism*
  • Disks Large Homolog 4 Protein
  • Endocytosis
  • Guanylate Kinases / metabolism*
  • Hippocampus / metabolism
  • Immunohistochemistry
  • Immunoprecipitation
  • Intracellular Signaling Peptides and Proteins / metabolism*
  • Membrane Proteins / metabolism*
  • Mice
  • Microscopy, Confocal
  • Nerve Tissue Proteins / metabolism*
  • Neurons / metabolism*
  • PDZ Domains
  • Patch-Clamp Techniques
  • Rats
  • Receptors, N-Methyl-D-Aspartate / metabolism*
  • Synapses / metabolism
  • Synaptic Membranes / metabolism*
  • Vesicular Transport Proteins / metabolism*

Substances

  • Adaptor Proteins, Signal Transducing
  • Disks Large Homolog 4 Protein
  • Dlg4 protein, mouse
  • Dlg4 protein, rat
  • Intracellular Signaling Peptides and Proteins
  • Membrane Proteins
  • Nerve Tissue Proteins
  • Receptors, N-Methyl-D-Aspartate
  • Vesicular Transport Proteins
  • rabphilin-3A
  • Guanylate Kinases
  • N-methyl D-aspartate receptor subtype 2A