Endocytosis and synaptic removal of NR3A-containing NMDA receptors by PACSIN1/syndapin1.
A key step in glutamatergic synapse maturation is the replacement of developmentally expressed N-methyl-D-aspartate receptors (NMDARs) with mature forms that differ in subunit composition, electrophysiological properties and propensity to elicit synaptic plasticity. However, the mechanisms underlying the removal and replacement of synaptic NMDARs are poorly understood. Here we demonstrate that NMDARs containing the developmentally regulated NR3A subunit undergo rapid endocytosis from the dendritic plasma membrane in cultured rat hippocampal neurons. This endocytic removal is regulated by PACSIN1/syndapin1, which directly and selectively binds the carboxy-terminal domain of NR3A through its NPF motifs and assembles a complex of proteins including dynamin and clathrin. Endocytosis of NR3A by PACSIN1 is activity dependent, and disruption of PACSIN1 function causes NR3A accumulation at synaptic sites. Our results reveal a new activity-dependent mechanism involved in the regulation of NMDAR expression at synapses during development, and identify a brain-specific endocytic adaptor that confers spatiotemporal and subunit specificity to NMDAR endocytosis.
Pérez-Otaño
Isabel
I
Department of Neurobiology, Duke University Medical Center, Box 3209, Durham, North Carolina 27710, USA. otano@unav.es
Luján
Rafael
R
Tavalin
Steven J
SJ
Plomann
Markus
M
Modregger
Jan
J
Liu
Xiao-Bo
XB
Jones
Edward G
EG
Heinemann
Stephen F
SF
Lo
Donald C
DC
Ehlers
Michael D
MD
D000818:Animals; D015153:Blotting, Western / Q000379:methods; D002478:Cells, Cultured; D003001:Cloning, Molecular / Q000379:methods; D004594:Electrophysiology / Q000379:methods; D004622:Embryo, Mammalian; D004705:Endocytosis / Q000187:drug effects / Q000502:physiology*; D005455:Fluorescent Antibody Technique / Q000379:methods; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006624:Hippocampus / Q000166:cytology; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods; D008562:Membrane Glycoproteins / Q000235:genetics / Q000502:physiology*; D016253:Microscopy, Immunoelectron / Q000379:methods; D009154:Mutation / Q000502:physiology; D016202:N-Methylaspartate / Q000494:pharmacology; D009474:Neurons / Q000166:cytology* / Q000187:drug effects; D011509:Proteoglycans / Q000235:genetics / Q000502:physiology*; D051381:Rats; D016194:Receptors, N-Methyl-D-Aspartate / Q000235:genetics / Q000378:metabolism*; D013569:Synapses / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D053667:Syndecans; D014162:Transfection / Q000379:methods; D020798:Two-Hybrid System Techniques
D008562:Membrane Glycoproteins; C464437:NR3A NMDA receptor; D011509:Proteoglycans; D016194:Receptors, N-Methyl-D-Aspartate; D053667:Syndecans; D049452:Green Fluorescent Proteins; D016202:N-Methylaspartate