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Endocytosis and synaptic removal of NR3A-containing NMDA receptors by PACSIN1/syndapin1

Nature Neuroscience volume 9, pages 611–621 (2006)Cite this article

Abstract

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.

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Figure 1: NMDARs containing NR3A subunits are inefficiently targeted to synapses and reside in intracellular compartments.
Figure 2: NR3A localizes to extrasynaptic and perisynaptic membrane domains.
Figure 3: Ultrastructural localization of NR3A at CA1 hippocampal synapses of adult rat using postembedding immunogold labeling.
Figure 4: NR3A-containing NMDARs undergo activity-dependent endocytosis in hippocampal neurons.
Figure 5: NR3A binds the neuron-specific endocytic adaptor PACSIN1/syndapin1.
Figure 6: PACSIN1 mediates NR3A endocytosis.
Figure 7: PACSIN1 reduces the functional surface expression of NR3A-containing triheteromeric NMDARs.
Figure 8: PACSIN1 mediates synaptic removal of NR3A.

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Acknowledgements

We thank J.F. Wesseling for intellectual support; K. Hawk, A. Zandueta, I. López-García, C. Zhang and H. Zhang for technical help; J.R. Naranjo and M. Palczewska for support with yeast assays; and G. Augustine, T. Blanpied, J. Hernandez, B. Philpot, A. Horton and R. Mooney for critical readings of this manuscript. This work was supported by grants from the National Alliance for Research on Schizophrenia and Depression, the Marie Curie International Reintegration Grant (IRG) Program and the Unión Temporal de Empresas project (UTE) at the Centro de Investigación Médica Aplicada (CIMA; I.P.O.), Deutsche Forschungsgemeinschaft (PL233/1-1, M.P.) and the Center for Molecular Medicine (CMMC) of the University of Cologne (TP78, M.P.). Research in D.C.L.'s laboratory is supported by grants from the US National Institutes of Health (NS32742). S.J.T. is supported by grants from the US National Institutes of Health (NS46661); R.L. by grants from Junta de Comunidades de Castilla-La Mancha (SAN-04-008-00, PAI05-040); M.D.E. by grants from the US National Institutes of Health (NS39402, MH64748), the American Heart Association, the Raymond and Beverley Sackler Foundation and the Ruth K. Broad Foundation; and S.F.H. by the US National Institutes of Health (R01 NS28709) and the McKnight and Adler Foundations. M.D.E is an Investigator of the Howard Hughes Medical Institute.

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Authors and Affiliations

  1. Department of Neurobiology, Duke University Medical Center, Box 3209, Durham, 27710, North Carolina, USA

    Isabel Pérez-Otaño, Donald C Lo & Michael D Ehlers

  2. Neuroscience Department, Cellular Neurobiology Laboratory, Centro de Investigación Médica Aplicada (CIMA), Universidad de Navarra, Pamplona, 31008, Spain

    Isabel Pérez-Otaño

  3. Departamento Ciencias Médicas, CRIB-Facultad de Medicina, Universidad de Castilla-La Mancha, Albacete, 02006, Spain

    Rafael Luján

  4. Department of Pharmacology, University of Tennessee Health Science Center, Memphis, 38163, Tennessee, USA

    Steven J Tavalin

  5. Center for Biochemistry and Center for Molecular Medicine, University of Cologne, Cologne, D-50931, Germany

    Markus Plomann & Jan Modregger

  6. Center for Neuroscience, University of California Davis, Davis, 95616, California, USA

    Xiao-Bo Liu & Edward G Jones

  7. Molecular Neurobiology Laboratory, The Salk Institute for Biological Studies, La Jolla, 92037, California, USA

    Stephen F Heinemann

  8. Howard Hughes Medical Institute, Duke University Medical Center, Durham, 27710, North Carolina, USA

    Michael D Ehlers

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Supplementary information

Supplementary Fig. 1

Internalized NR3A is transported to early endosomes. (PDF 913 kb)

Supplementary Fig. 2

Expression of NR3A and PACSIN1 in rat brain. (PDF 1096 kb)

Supplementary Fig. 3

Expression of PACSIN1 in hippocampal neurons. (PDF 2455 kb)

Supplementary Fig. 4

Proposed model for activity-dependent synaptic removal of NR3A and regulation of NMDAR maturation during development. (PDF 788 kb)

Supplementary Methods (PDF 148 kb)

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Pérez-Otaño, I., Luján, R., Tavalin, S. et al. Endocytosis and synaptic removal of NR3A-containing NMDA receptors by PACSIN1/syndapin1. Nat Neurosci 9, 611–621 (2006). https://doi.org/10.1038/nn1680

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