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Volume 16, Number 14, Issue of July 15, 1996 pp. 4457-4467
Copyright ©1996 Society for NeuroscienceOrganization of AMPA Receptor Subunits at a Glutamate Synapse: A Quantitative Immunogold Analysis of Hair Cell Synapses in the Rat Organ of Corti
Received Feb. 29, 1996; revised April 18, 1996; accepted April 24, 1996.
Atsushi Matsubara1, 2, Jon H. Laake1, Svend Davanger1, Shin-ichi Usami2, and Ole P. Ottersen1 1 Department of Anatomy, Institute of Basic Medical Sciences, University of Oslo, P.O. Box 1105 Blindern, N-0317 Oslo, Norway, and 2 Department of Otorhinolaryngology, Hirosaki University School of Medicine, Hirosaki, 036 Japan
Sensitive and high-resolution immunocytochemical procedures were used to investigate the spatial organization of AMPA receptor subunits (GluR1-4) at the synapse between the inner hair cells and the afferent dendrites in the rat organ of Corti. This is a synapse with special functional properties and with a presynaptic dense body that defines the center of the synapse and facilitates its morphometric analysis. A quantitative postembedding immunocytochemical analysis was performed on specimens that had been embedded in a metachrylate resin at low temperature after freeze substitution. Single- and double-labeling procedures indicated that GluR2/3 and GluR4 subunits were colocalized throughout the postsynaptic density, with a maximum distance of 300 nm from the presynaptic body and with higher concentrations peripherally than centrally. No receptor immunolabeling was found at extrasynaptic membranes, but some GluR4 subunits appeared to be expressed presynaptically. The synapses between outer hair cells and afferent dendrites were devoid of labeling. The present data indicate that AMPA receptor subunits are inserted into the postsynaptic membrane in a very precise manner and that their density increases on moving away from the center of the synapse.
Key words: AMPA; pre- and postsynaptic receptors; immunocytochemistry; immunogold; silver enhancement; organ of Corti; rat
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