Immunohistochemical localization of N-methyl-d-aspartate and α-amino-3-hydroxy-5-methyl-4-isoxazolepropionate receptor subunits in the substantia nigra pars compacta of the rat

Ionotropic glutamate receptors in the substantia nigra pars compacta regulate the activity of dopamine neurons. We have used dual-label immunofluoresence and confocal laser microscopy to study the localization of subunits of two types of ionotropic receptors within the substantia nigra pars compacta of the rat. Immunostaining for N-methyl-D-aspartate receptor 1 and glutamate receptor 2/3 was prominent in the soma and proximal dendrites of all tyrosine hydroxylase-immunopositive cells, while only low amounts of N-methyl-D-aspartate receptor 2A and N-methyl-D-aspartate receptor 2B were present. Selective antibodies were used to determine the isoforms of N-methyl-D-aspartate receptor 1 present. Immunostaining for the N1, C1 and C2 variably spliced segments of N-methyl-D-aspartate receptor 1 were scarce in the substantia nigra pars compacta, while immunoreactivity for the alternative C2' terminus of N-methyl-D-aspartate receptor 1 was quite abundant. Staining for glutamate receptor 1 was heterogeneous; about half of the tyrosine hydroxylase immunopositive cells stained intensely, while the other half were immunonegative. The glutamate receptor 1-stained cells were concentrated in the ventral tier of the substantia nigra pars compacta. Glutamate receptor 4 was not found in tyrosine hydroxylase-immunopositive cells within the substantia nigra pars compacta. Together, these data demonstrate that dopaminergic neurons in the substantia nigra pars compacta express primarily glutamate receptor 1, glutamate receptor 2/3 and N-methyl-D-aspartate receptor 1 isoforms containing the alternative C2' terminus.

[1]  R. Huganir,et al.  Regulation of NMDA receptor phosphorylation by alternative splicing of the C-terminal domain , 1993, Nature.

[2]  S. Nakanishi,et al.  Molecular cloning and characterization of the rat NMDA receptor , 1991, Nature.

[3]  G Chouvet,et al.  Tonic Activation of NMDA Receptors Causes Spontaneous Burst Discharge of Rat Midbrain Dopamine Neurons In Vivo , 1993, The European journal of neuroscience.

[4]  R. Huganir,et al.  Inactivation of NMDA Receptors by Direct Interaction of Calmodulin with the NR1 Subunit , 1996, Cell.

[5]  D. Laurie,et al.  Ligand affinities at recombinant N-methyl-D-aspartate receptors depend on subunit composition. , 1994, European journal of pharmacology.

[6]  Y. Jan,et al.  Changing subunit composition of heteromeric NMDA receptors during development of rat cortex , 1994, Nature.

[7]  M. Iadarola,et al.  NMDAR1 Glutamate Receptor Subunit Isoforms in Neostriatal, Neocortical, and Hippocampal Nitric Oxide Synthase Neurons , 1998, The Journal of Neuroscience.

[8]  A. Lees,et al.  Ageing and Parkinson's disease: substantia nigra regional selectivity. , 1991, Brain : a journal of neurology.

[9]  S. Heinemann,et al.  Zinc potentiates agonist-lnduced currents at certain splice variants of the NMDA receptor , 1993, Neuron.

[10]  R. Huganir,et al.  AMPA glutamate receptor subunits are differentially distributed in rat brain , 1993, Neuroscience.

[11]  S. Heinemann,et al.  Control of proton sensitivity of the NMDA receptor by RNA splicing and polyamines. , 1995, Science.

[12]  M. Bennett,et al.  Alternatively spliced isoforms of the NMDARI receptor subunit , 1995, Trends in Neurosciences.

[13]  M. Tohyama,et al.  The differential expression patterns of messenger RNAs encoding non-N-methyl-d-aspartate glutamate receptor subunits (GluR1–4) in the rat brain , 1993, Neuroscience.

[14]  J. Morrison,et al.  Regional, cellular, and ultrastructural distribution of N-methyl-D-aspartate receptor subunit 1 in monkey hippocampus. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[15]  L. Meltzer,et al.  Evidence for N-methyl-d-aspartate and AMPA subtypes of the glutamate receptor on substantia nigra dopamine neurons: Possible preferential role for N-methyl-d-aspartate receptors , 1995, Neuroscience.

[16]  P. Seeburg,et al.  Regional and developmental heterogeneity in splicing of the rat brain NMDAR1 mRNA , 1994, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[17]  M. Beal.  Mechanisms of excitotoxicity in neurologic diseases , 1992, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[18]  S. Nakanishi,et al.  Structures and properties of seven isoforms of the NMDA receptor generated by alternative splicing. , 1992, Biochemical and biophysical research communications.

[19]  R. Petralia,et al.  Light and electron immunocytochemical localization of AMPA‐selective glutamate receptors in the rat brain , 1992, The Journal of comparative neurology.

[20]  Bert Sakmann,et al.  Heteromeric NMDA Receptors: Molecular and Functional Distinction of Subtypes , 1992, Science.

[21]  M. Bennett,et al.  Splice variants of the N-methyl-D-aspartate receptor NR1 identify domains involved in regulation by polyamines and protein kinase C. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[22]  V. Pickel,et al.  Comparative ultrastructural localization of the NMDAR1 glutamate receptor in the rat basolateral amygdala and bed nucleus of the stria terminalis , 1995, The Journal of comparative neurology.

[23]  J. Penney,et al.  Excitatory amino acid binding sites in the basal ganglia of the rat: A quantitative autoradiographic study , 1992, Neuroscience.

[24]  S. Heinemann,et al.  Cloned glutamate receptors. , 1994, Annual review of neuroscience.

[25]  Y. Smith,et al.  AMPA and NMDA Glutamate Receptor Subunits in Midbrain Dopaminergic Neurons in the Squirrel Monkey: An Immunohistochemical andIn Situ Hybridization Study , 1997, The Journal of Neuroscience.

[26]  S. Hoffman,et al.  Protection against malaria by vaccination with sporozoite surface protein 2 plus CS protein. , 1991, Science.

[27]  B. Sakmann,et al.  Structural determinants of ion flow through recombinant glutamate receptor channels , 1991, Science.

[28]  M. Sheng,et al.  Heteromultimerization and NMDA Receptor-Clustering Activity of Chapsyn-110, a Member of the PSD-95 Family of Proteins , 1996, Neuron.

[29]  R. Axel,et al.  Alternative splicing generates functionally distinct N-methyl-D-aspartate receptors. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[30]  F. Gonon.  Nonlinear relationship between impulse flow and dopamine released by rat midbrain dopaminergic neurons as studied by in vivo electrochemistry , 1988, Neuroscience.

[31]  K. Moriyoshi,et al.  Molecular characterization of the family of the N-methyl-D-aspartate receptor subunits. , 1993, The Journal of biological chemistry.

[32]  H. Tohgi,et al.  [Aging and Parkinson's disease]. , 1991, No to shinkei = Brain and nerve.

[33]  D. Monaghan,et al.  NR1 and NR2 subunit contributions to N-methyl-D-aspartate receptor channel blocker pharmacology. , 1997, The Journal of pharmacology and experimental therapeutics.

[34]  C. Saper,et al.  Organization of atriopeptin‐like immunoreactive neurons in the central nervous system of the rat , 1986, The Journal of comparative neurology.

[35]  Mary B. Kennedy,et al.  The postsynaptic density at glutamatergic synapses , 1997, Trends in Neurosciences.

[36]  R. Wenthold,et al.  Immunochemical characterization of the non-NMDA glutamate receptor using subunit-specific antibodies. Evidence for a hetero-oligomeric structure in rat brain. , 1992, The Journal of biological chemistry.

[37]  J. Penney,et al.  Organization of N‐methyl‐D‐aspartate glutamate receptor gene expression in the basal ganglia of the rat , 1994, The Journal of comparative neurology.

[38]  R. Wenthold,et al.  The NMDA receptor subunits NR2A and NR2B show histological and ultrastructural localization patterns similar to those of NR1 , 1994, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[39]  B. Sakmann,et al.  Developmental and regional expression in the rat brain and functional properties of four NMDA receptors , 1994, Neuron.

[40]  P. Seeburg,et al.  Domain interaction between NMDA receptor subunits and the postsynaptic density protein PSD-95. , 1995, Science.

[41]  R. North,et al.  Burst firing in dopamine neurons induced by N-methyl-D-aspartate: role of electrogenic sodium pump. , 1992, Science.

[42]  T. Verdoorn,et al.  N-methyl-D-aspartate receptors: different subunit requirements for binding of glutamate antagonists, glycine antagonists, and channel-blocking agents. , 1994, Molecular pharmacology.

[43]  C. Aoki,et al.  NMDA-R1 subunit of the cerebral cortex co-localizes with neuronal nitric oxide synthase at pre- and postsynaptic sites and in spines , 1997, Brain Research.

[44]  D. Bredt,et al.  Interaction of Nitric Oxide Synthase with the Postsynaptic Density Protein PSD-95 and α1-Syntrophin Mediated by PDZ Domains , 1996, Cell.

[45]  P. Follesa,et al.  Chronic Ethanol-mediated Up-regulation of the N-Methyl-D-aspartate Receptor Polypeptide Subunits in Mouse Cortical Neurons in Culture* , 1996, The Journal of Biological Chemistry.

[46]  D. Standaert,et al.  Immunohistochemical localization of metabotropic glutamate receptors mGluR1a and mGluR2/3 in the rat basal ganglia , 1998, The Journal of comparative neurology.

[47]  S. Heinemann,et al.  Ca2+ permeability of KA-AMPA--gated glutamate receptor channels depends on subunit composition , 1991, Science.

[48]  B. Sakmann,et al.  Divalent ion permeability of AMPA receptor channels is dominated by the edited form of a single subunit , 1992, Neuron.

[49]  T. Dawson,et al.  Cellular and subcellular localization of NMDA-R1 subunit immunoreactivity in the visual cortex of adult and neonatal rats , 1994, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[50]  D. Clark,et al.  Iontophoretically administered drugs acting at the N‐methyl‐D‐aspartate receptor modulate burst firing in A9 dopamine neurons in the rat , 1992, Synapse.