Expression of AMPA, kainate, and NMDA receptor subunits in cochlear and vestibular ganglia

Glutamate is believed to be the principal afferent neurotransmitter in the peripheral auditory and vestibular systems. In this report, we present a comprehensive molecular analysis of ionotropic glutamate receptor gene expression in the cochlear and vestibular ganglia of the rat. Fourteen glutamate receptor subunits were studied: GluR1–4 (including flip and flop variants), GluR5–7, KA1&2, NR1, and NR2A-D. Reverse transcription of RNA followed by DNA amplification with the polymerase chain reaction was used for the initial analysis. Immunocytochemistry and in situ hybridization with subunit-specific oligonucleotides were subsequently used for cellular localization of receptor expression. AMPA (GluR2–4), kainate (GluR5&6 and KA1&2), and NMDA receptor (NR1 and NR2A-D) subunit expression was detected. Based on the relative amounts of mRNA detected by in situ hybridization, the predominant receptors expressed by cochlear and vestibular ganglion cells appear to be GluR2, GluR3, GluR4, GluR5, and NR1. At a moderate level were GluR6, NR2B, and NR2D. KA1, KA2, NR2A, and NR2C mRNAs were also expressed in ganglion cells, but at lower levels. Only the AMPA receptor subunit GluR1 and the kainate receptor subunit GluR7 were not found to be expressed in vestibulocochlear neurons. These studies suggest that functional AMPA, kainate, and NMDA receptors are present at the hair cell/vestibulocochlear nerve synapse.

[1]  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.

[2]  R. Petralia,et al.  Light and electron microscopic immunocytochemical localization of AMPA‐selective glutamate receptors in the rat spinal cord , 1994, The Journal of comparative neurology.

[3]  M Tohyama,et al.  Expression of NMDA receptor subunit mRNA in the vestibular ganglion of the rat and guinea-pig. , 1994, Neuroreport.

[4]  J. Puel,et al.  Excitatory amino acid antagonists protect cochlear auditory neurons from excitotoxicity , 1994, The Journal of comparative neurology.

[5]  R. Wenthold,et al.  Light and electron microscope distribution of the NMDA receptor subunit NMDAR1 in the rat nervous system using a selective anti-peptide antibody , 1994, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[6]  R. Petralia,et al.  Biochemical and assembly properties of GluR6 and KA2, two members of the kainate receptor family, determined with subunit-specific antibodies. , 1994, The Journal of biological chemistry.

[7]  M. Mayer,et al.  Selective modulation of desensitization at AMPA versus kainate receptors by cyclothiazide and concanavalin A , 1993, Neuron.

[8]  G. Uhl,et al.  Expression and novel subunit isoforms of glutamate receptor genes GluR5 and GluR6. , 1993, Neuroreport.

[9]  M. Mayer,et al.  Differential modulation by cyclothiazide and concanavalin A of desensitization at native alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid- and kainate-preferring glutamate receptors. , 1993, Molecular pharmacology.

[10]  R. Petralia,et al.  Expression of AMPA-selective glutamate receptor subunits in morphologically defined neurons of the mammalian cochlear nucleus , 1993, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[11]  M. Eybalin,et al.  Neurotransmitters and neuromodulators of the mammalian cochlea. , 1993, Physiological reviews.

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

[13]  W Wisden,et al.  The rat delta‐1 and delta‐2 subunits extend the excitatory amino acid receptor family , 1993, FEBS letters.

[14]  S. Safieddine,et al.  Co-expression of NMDA and AMPA/kainate receptor mRNAs in cochlear neurones. , 1992, Neuroreport.

[15]  Masahiko Watanabe,et al.  Cloning and expression of the ε4 subunit of the NMDA receptor channel , 1992 .

[16]  S. Nakanishi.  Molecular diversity of glutamate receptors and implications for brain function. , 1992, Science.

[17]  J. Rossier,et al.  AMPA receptor subunits expressed by single purkinje cells , 1992, Neuron.

[18]  K. Sakimura,et al.  Molecular diversity of the NMDA receptor channel , 1992, Nature.

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

[20]  T Kawase,et al.  Spatial organization of the auditory nerve according to spontaneous discharge rate , 1992, The Journal of comparative neurology.

[21]  M. Yamazaki,et al.  Functional characterization of a heteromeric NMDA receptor channel expressed from cloned cDNAs , 1992, Nature.

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

[23]  B. Sakmann,et al.  The KA-2 subunit of excitatory amino acid receptors shows widespread expression in brain and forms ion channels with distantly related subunits , 1992, Neuron.

[24]  B. Sakmann,et al.  A glutamate receptor channel with high affinity for domoate and kainate. , 1992, The EMBO journal.

[25]  M. Yamazaki,et al.  Molecular cloning of a cDNA encoding a novel member of the mouse glutamate receptor channel family. , 1992, Biochemical and biophysical research communications.

[26]  L. Vyklický,et al.  Molecular cloning and development analysis of a new glutamate receptor subunit isoform in cerebellum , 1992, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[27]  K. Sakimura,et al.  Primary structure and expression of the γ 2 subunit of the glutamate receptor channel selective for kainate , 1992, Neuron.

[28]  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.

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

[30]  A. Ryan,et al.  Preservation of mRNA during in situ hybridization in the cochlea , 1991, Hearing Research.

[31]  G. Moonen,et al.  Kainate and NMDA toxicity for cultured developing and adult rat spiral ganglion neurons: further evidence for a glutamatergic excitatory neurotransmission at the inner hair cell synapse , 1991, Brain Research.

[32]  S. Heinemann,et al.  Cloning of a cDNA for a glutamate receptor subunit activated by kainate but not AMPA , 1991, Nature.

[33]  P. Seeburg,et al.  Cloning of a putative high-affinity kainate receptor expressed predominantly in hippocampal CA3 cells , 1991, Nature.

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

[35]  H. Monyer,et al.  Glutamate-operated channels: Developmentally early and mature forms arise by alternative splicing , 1991, Neuron.

[36]  D. Ryugo,et al.  Neurofilament antibodies and spiral ganglion neurons of the mammalian cochlea , 1991, The Journal of comparative neurology.

[37]  S. Komune,et al.  Excitatory amino acid response in isolated spiral ganglion cells of guinea pig cochlea. , 1991, Journal of neurophysiology.

[38]  J. Puel,et al.  Electrophysiological evidence for the presence of NMDA receptors in the guinea pig cochlea , 1991, Hearing Research.

[39]  S. Heinemann,et al.  Cloning of a novel glutamate receptor subunit, GluR5: Expression in the nervous system during development , 1990, Neuron.

[40]  B. Sakmann,et al.  Flip and flop: a cell-specific functional switch in glutamate-operated channels of the CNS. , 1990, Science.

[41]  J. E. Huettner.  Glutamate receptor channels in rat DRG neurons: Activation by kainate and quisqualate and blockade of desensitization by con A , 1990, Neuron.

[42]  D. Choi,et al.  Glutamate neurotoxicity and diseases of the nervous system , 1988, Neuron.

[43]  E. Mroz,et al.  Neuroactive substances in inner ear extracts , 1987, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[44]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[45]  P. Ascher,et al.  Glycine potentiates the NMDA response in cultured mouse brain neurons , 1987, Nature.

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

[47]  P. Seeburg,et al.  Glutamate receptor channels: novel properties and new clones. , 1992, Trends in pharmacological sciences.

[48]  J. Puel,et al.  Alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid electrophysiological and neurotoxic effects in the guinea-pig cochlea. , 1991, Neuroscience.

[49]  R. Bobbin,et al.  Comparative actions of quisqualate and N-methyl-D-aspartate, excitatory amino acid agonists, on guinea-pig cochlear potentials. , 1986, Comparative biochemistry and physiology. C, Comparative pharmacology and toxicology.