Immunocytochemical localization of the postsynaptic density protein PSD-95 in the mammalian retina.
Synapse-associated proteins are the scaffold for the selective aggregation of ion channels at synapses; they provide the link to cytoskeletal elements and possibly are involved with the regulation of synaptic efficacy by electrical activity. The localization of the postsynaptic density protein PSD-95 was studied in different mammalian retinae (rat, monkey, and tree shrew) by using immunocytochemical methods. Immunofluorescence for PSD-95 was most prominent in the outer plexiform layer (OPL). The axon terminals of rods and cones, the rod spherules and cone pedicles, were strongly labeled. Electron microscopy, using preembedding immunocytochemistry, showed PSD-95 localized presynaptically within the photoreceptor terminals. Distinct PSD-95 labeling was also present in the inner plexiform layer (IPL). It had a punctate appearance suggesting the synaptic clustering of PSD-95 in the IPL. Electron microscopy showed that PSD-95 was concentrated in processes that were postsynaptic at bipolar cell ribbon synapses (dyads). As a rule, only one of the two postsynaptic members of the dyad was labeled for PSD-95. Double-labeling experiments were performed for PSD-95 and for SAP 102 or PSD-93, respectively, two other members of the family of synapse-associated proteins. All three were found to be colocalized in the synaptic hot spots in the IPL. In the OPL, however, PSD-95 and PSD-93 were found presynaptically, whereas SAP 102 was located postsynaptically at photoreceptor synapses. Double-labeling experiments also were performed for PSD-95 and for the NR1 subunit of the NMDA receptor. They were found to be colocalized in synaptic hot spots in the IPL.
Koulen
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Max-Planck-Institut für Hirnforschung, D-60528 Frankfurt, Germany.
Fletcher
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EL
Craven
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Bredt
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DS
Wässle
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H
D000818:Animals; D000911:Antibodies, Monoclonal; D000076122:Disks Large Homolog 4 Protein; D047908:Intracellular Signaling Peptides and Proteins; D008253:Macaca mulatta; D008565:Membrane Proteins; D008854:Microscopy, Electron; D009419:Nerve Tissue Proteins / Q000032:analysis* / Q000276:immunology; D009479:Neuropeptides / Q000032:analysis / Q000276:immunology; D051381:Rats; D011919:Rats, Inbred Strains; D016194:Receptors, N-Methyl-D-Aspartate / Q000032:analysis / Q000276:immunology; D017949:Retinal Cone Photoreceptor Cells / Q000737:chemistry* / Q000648:ultrastructure; D017948:Retinal Rod Photoreceptor Cells / Q000737:chemistry* / Q000648:ultrastructure; D013570:Synaptic Membranes / Q000737:chemistry*; D014417:Tupaia
D000911:Antibodies, Monoclonal; D000076122:Disks Large Homolog 4 Protein; C101195:Dlg3 protein, rat; C501449:Dlg4 protein, rat; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; C501451:Mpp2 protein, rat; D009419:Nerve Tissue Proteins; D009479:Neuropeptides; D016194:Receptors, N-Methyl-D-Aspartate; C033652:postsynaptic density proteins