Figure 1

Supporting cell (SC) expression of miR-182 in Tg^1MDW/1MDW mice at P18. (A–D) Dual ISH/IHC fluorescence imaging of whole mount organ of Corti (OC) from WT (A,C) and Tg^1MDW/1MDW (B,D) P18 littermates. Cy5 fluorescence of tyramide enhanced miR-182 ISH labeling shows nuclear/cytoplasmic staining in hair cells (HCs A,B) and definitive Tg^1MDW/1MDW OC SC nuclear staining, including Deiters’, inner pillar, outer pillar, inner phalangeal and inner border cells (B, arrowheads). SC labeling with miR-182 was found in spiral limbus cells and myelinated Schwann cells of spiral ganglion neurons (B, arrows). Alexa 546 immunofluorescence of MYO6 positive HCs (C,D). (E) Quantitative increase in miR-183 cluster in P18 Tg^1MDW/1MDW total cochlear RNA by RT-PCR. Using ABI Taqman assays, quantitative RT-PCR was performed in Tg^1MDW/1MDW versus WT littermates (n = 3). The miR-183, miR-96 and miR-182 levels were normalized to Sno135. The results quantitate statistically significant (ΔCT values, 2 sample t-test, P < 0.001) increases in miR-182 (2.9 fold), miR-96 (2.7 fold) and miR-183 (2.2 fold) in Tg^1MDW/1MDW cochlea at P18. These data are consistent with a transgenic misexpression of these neurosensory miRNAs via the core human GFAP promoter and validate microarray data. Scale bar: 20 μM (A–D).