OHC synaptic immunopuncta.

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OHC synaptic immunopuncta. OHC synaptic immunopuncta. Confocal z-stack of OHCs in the middle turn of the organ of Corti from an adult rat viewed from the endolymphatic surface. A, Immunolabeling with the postsynaptic density proteins PSD-95 (red channel) and Shank (green channel) show an interconnected series of puncta along the base of the OHCs. PSD-95 and Shank puncta are closely coincident (magnified insert, x- to y-plane). B, Immunolabeling with the presynaptic ribbon marker CtBP2 (red) and postsynaptic marker Shank (green). C, Immunolabel for CtBP2 (red) and PSD-95 (green). D, Immunolabel for CtPB2 (red) and Homer (green). Magnified inserts (x- to y-plane) in each case show more extensive postsynaptic density distribution than presynaptic ribbon label. E, Presynaptic and postsynaptic immunopuncta at the OHCs. There were significantly fewer CtBP2 or GluA2 puncta than postsynaptic density puncta (PSD-95, Shank, or Homer; one way-ANOVA, p = 0.01; Bonferroni’s multiple comparison test; n = 3-7; 72-168 OHCs). F, Thumbnails of the base of individual OHCs immunolabeled for CtBP2 (red channel) and Shank (green channel). Many Shank immunopuncta had no associated CtBP2 puncta. G, Thumbnails of the base of individual OHCs immunolabeled for GluA2 (red channel) and Shank (green channel). Many of the Shank immunopuncta had no associated GluA2 puncta. H, Percentage juxtaposition of the CtBP2 and PSDs. The ratio of PSD-95, Shank, or Homer puncta juxtaposed to CTBP2 was significantly smaller than the ratio of CTBP2 puncta juxtaposed to the postsynaptic density proteins (one way-ANOVA, p ≤ 0.01; Bonferroni’s multiple comparison test; i.e., many PSDs had no ribbon). Scale bars: wide view, 5 µm; magnified inserts and thumbnails, 2.5 µm. I, Schematic drawing of OHC and IHC synapses. At the IHC afferent synapse, CtBP2/GluA2 relates closely in number to postsynaptic density markers. At the OHC afferent synapse, only a subset of postsynaptic density proteins relates to CtBP2/GluA2 synaptic markers. Rodrigo Martinez-Monedero et al. eneuro 2016;3:ENEURO.0078-16.2016 ©2016 by Society for Neuroscience