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Journal of Neuroscience, Vol 14, 667-696, Copyright © 1994 by Society for Neuroscience

ARTICLE

Light and electron microscope distribution of the NMDA receptor subunit NMDAR1 in the rat nervous system using a selective anti-peptide antibody

RS Petralia, N Yokotani and RJ Wenthold
Laboratory of Neurochemistry, NIDCD, NIH, Bethesda, Maryland 20892.

NMDA receptors play key roles in synaptic plasticity and neuronal development, and may be involved in learning, memory, and compensation following injury. A polyclonal antibody that recognizes four of seven splice variants of NMDAR1 was made using a C-terminus peptide (30 amino acid residues). NMDAR1 is the major NMDA receptor subunit, found in most or all NMDA receptor complexes. On immunoblots, this antibody labeled a single major band migrating at M(r) = 120,000. The antibody did not cross-react with extracts from transfected cells expressing other glutamate receptor subunits, nor did it label non-neuronal tissues. Immunostained vibratome sections of rat tissue showed labeling in many neurons in most structures in the brain, as well as in the cervical spinal cord, dorsal root and vestibular ganglia, and in pineal and pituitary glands. Staining was moderate to dense in the olfactory bulb, neocortex, striatum, some thalamic and hypothalamic nuclei, the colliculi, and many reticular, sensory, and motor neurons of the brainstem and spinal cord. The densest stained cells included the pyramidal and hilar neurons of the CA3 region of the hippocampus, Purkinje cells of the cerebellum, supraoptic and magnocellular paraventricular neurons of the hypothalamus, inferior olive, red nucleus, lateral reticular nucleus, peripheral dorsal cochlear nucleus, and motor nuclei of the lower brainstem and spinal cord. Ultrastructural localization of immunostaining was examined in the hippocampus, cerebral cortex, and cerebellar cortex. The major staining was in postsynaptic densities apposed by unstained presynaptic terminals with round or mainly round vesicles, and in associated dendrites. The pattern of staining matched that of previous in situ hybridization but differed somewhat from that of binding studies, implying that multiple types of NMDA receptors exist. Comparison with previous studies of localization of other glutamate receptor types revealed that NMDAR1 may colocalize with these other types in many neurons throughout the nervous system.

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				M. Wyszynski, V. Kharazia, R. Shanghvi, A. Rao, A. H. Beggs, A. M. Craig, R. Weinberg, and M. Sheng Differential Regional Expression and Ultrastructural Localization of alpha -Actinin-2, a Putative NMDA Receptor-Anchoring Protein, in Rat Brain J. Neurosci., February 15, 1998; 18(4): 1383 - 1392. [Abstract] [Full Text] [PDF]

				Y.-X. Wang, R. J. Wenthold, O. P. Ottersen, and R. S. Petralia Endbulb Synapses in the Anteroventral Cochlear Nucleus Express a Specific Subset of AMPA-Type Glutamate Receptor Subunits J. Neurosci., February 1, 1998; 18(3): 1148 - 1160. [Abstract] [Full Text] [PDF]

				G. Yang, C. Iadecola, and F. M. Faraci Activation of Cerebellar Climbing Fibers Increases Cerebellar Blood Flow : Role of Glutamate Receptors, Nitric Oxide, and cGMP • Editorial Comment: Role of Glutamate Receptors, Nitric Oxide, and cGMP Stroke, February 1, 1998; 29(2): 499 - 508. [Abstract] [Full Text] [PDF]

				M. D. Ehlers, E. T. Fung, R. J. O'Brien, and R. L. Huganir Splice Variant-Specific Interaction of the NMDA Receptor Subunit NR1 with Neuronal Intermediate Filaments J. Neurosci., January 15, 1998; 18(2): 720 - 730. [Abstract] [Full Text] [PDF]

				G. Bai, D. D. Norton, M. S. Prenger, and J. W. Kusiak Single-stranded DNA-binding Proteins and Neuron-restrictive Silencer Factor Participate in Cell-specific Transcriptional Control of the NMDAR1 Gene J. Biol. Chem., January 9, 1998; 273(2): 1086 - 1091. [Abstract] [Full Text] [PDF]

				M. Mock, C. Schwarz, and P. Thier Electrophysiological Properties of Rat Pontine Nuclei Neurons In Vitro II. Postsynaptic Potentials J Neurophysiol, December 1, 1997; 78(6): 3338 - 3350. [Abstract] [Full Text] [PDF]

				F. S. Livingston and R. Mooney Development of Intrinsic and Synaptic Properties in a Forebrain Nucleus Essential to Avian Song Learning J. Neurosci., December 1, 1997; 17(23): 8997 - 9009. [Abstract] [Full Text] [PDF]

				A. L. Kirchgessner, M.-T. Liu, and F. Alcantara Excitotoxicity in the Enteric Nervous System J. Neurosci., November 15, 1997; 17(22): 8804 - 8816. [Abstract] [Full Text] [PDF]

				D. W. Brann and V. B. Mahesh Excitatory Amino Acids: Evidence for a Role in the Control of Reproduction and Anterior Pituitary Hormone Secretion Endocr. Rev., October 1, 1997; 18(5): 678 - 700. [Abstract] [Full Text]

				R. J. O'Brien, A. L. Mammen, S. Blackshaw, M. D. Ehlers, J. D. Rothstein, and R. L. Huganir The Development of Excitatory Synapses in Cultured Spinal Neurons J. Neurosci., October 1, 1997; 17(19): 7339 - 7350. [Abstract] [Full Text] [PDF]

				S. Safieddine and R. J. Wenthold The Glutamate Receptor Subunit delta 1 Is Highly Expressed in Hair Cells of the Auditory and Vestibular Systems J. Neurosci., October 1, 1997; 17(19): 7523 - 7531. [Abstract] [Full Text] [PDF]

				G. D. Funk, S. M. Johnson, J. C. Smith, X.-W. Dong, J. Lai, and J. L. Feldman Functional Respiratory Rhythm Generating Networks in Neonatal Mice Lacking NMDAR1 Gene J Neurophysiol, September 1, 1997; 78(3): 1414 - 1420. [Abstract] [Full Text] [PDF]

				M.-T. Liu, J. D. Rothstein, M. D. Gershon, and A. L. Kirchgessner Glutamatergic Enteric Neurons J. Neurosci., June 15, 1997; 17(12): 4764 - 4784. [Abstract] [Full Text] [PDF]

				K. N. Gracy, A. L. Svingos, and V. M. Pickel Dual Ultrastructural Localization of µ-Opioid Receptors and NMDA-Type Glutamate Receptors in the Shell of the Rat Nucleus Accumbens J. Neurosci., June 15, 1997; 17(12): 4839 - 4848. [Abstract] [Full Text] [PDF]

				A. S. Leonard and J. W. Hell Cyclic AMP-dependent Protein Kinase and Protein Kinase C Phosphorylate N-Methyl-D-aspartate Receptors at Different Sites J. Biol. Chem., May 2, 1997; 272(18): 12107 - 12115. [Abstract] [Full Text] [PDF]

				K. Gottmann, A. Mehrle, G. Gisselmann, and H. Hatt Presynaptic Control of Subunit Composition of NMDA Receptors Mediating Synaptic Plasticity J. Neurosci., April 15, 1997; 17(8): 2766 - 2774. [Abstract] [Full Text] [PDF]

				A. H. Gazzaley, D. L. Benson, G. W. Huntley, and J. H. Morrison Differential Subcellular Regulation of NMDAR1 Protein and mRNA in Dendrites of Dentate Gyrus Granule Cells after Perforant Path Transection J. Neurosci., March 15, 1997; 17(6): 2006 - 2017. [Abstract] [Full Text] [PDF]

				M. J. Schell, R. O. Brady Jr., M. E. Molliver, and S. H. Snyder D-Serine as a Neuromodulator: Regional and Developmental Localizations in Rat Brain Glia Resemble NMDA Receptors J. Neurosci., March 1, 1997; 17(5): 1604 - 1615. [Abstract] [Full Text] [PDF]

				M. Paquet, M. Tremblay, J.-J. Soghomonian, and Y. Smith AMPA and NMDA Glutamate Receptor Subunits in Midbrain Dopaminergic Neurons in the Squirrel Monkey: An Immunohistochemical and In Situ Hybridization Study J. Neurosci., February 15, 1997; 17(4): 1377 - 1396. [Abstract] [Full Text] [PDF]

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