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The Journal of Neuroscience, June 15, 2000, 20(12):4573-4581

C-Terminal Truncation of NR2A Subunits Impairs Synaptic But Not Extrasynaptic Localization of NMDA Receptors

Frank Steigerwald1, Torsten W. Schulz1, Leslie T. Schenker2, Mary B. Kennedy2, Peter H. Seeburg1, and Georg Köhr1

1 Max-Planck-Institute for Medical Research, Molecular Neurobiology, D-69120 Heidelberg, Germany, and 2 Division of Biology, California Institute of Technology, Pasadena, California 91125

NMDA receptors interact via the extended intracellular C-terminal domain of the NR2 subunits with constituents of the postsynaptic density for purposes of retention, clustering, and functional regulation at central excitatory synapses. To examine the role of the C-terminal domain of NR2A in the synaptic localization and function of NR2A-containing NMDA receptors in hippocampal Schaffer collateral-CA1 pyramidal cell synapses, we analyzed mice which express NR2A only in its C-terminally truncated form. In CA1 cell somata, the levels, activation, and deactivation kinetics of extrasynaptic NMDA receptor channels were comparable in wild-type and mutant NR2ADelta C/Delta C mice. At CA1 cell synapses, however, the truncated receptors were less concentrated than their full-length counterparts, as indicated by immunodetection in cultured neurons, synaptosomes, and postsynaptic densities. In the mutant, the NMDA component of evoked EPSCs was reduced in a developmentally progressing manner and was even more reduced in miniature EPSCs (mEPSCs) elicited by spontaneous glutamate release. Moreover, pharmacologically isolated NMDA currents evoked by synaptic stimulation had longer latencies and displayed slower rise and decay times, even in the presence of an NR2B-specific antagonist. These data strongly suggest that the C-terminal domain of NR2A subunits is important for the precise synaptic arrangement of NMDA receptors.

Key words: immunocytochemistry; Western blotting; patch clamp; hippocampal culture and slice; mice expressing C-terminally truncated NR2A subunits; nucleated patches; evoked EPSCs; miniature currents; NR2B-specific antagonists (CP-101,606)

Copyright © 2000 Society for Neuroscience  0270-6474/00/20124573-09$05.00/0


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