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. 2021 Aug 25;10(9):1351. doi: 10.3390/antiox10091351

Figure 3.

Figure 3

Comparative hearing evaluation, cochlear cytoarchitecture and organ of Corti degeneration. (A) Audiograms representing click and tone burst stimuli (4, 8, 16, 24 and 32 kHz) auditory brainstem response (ABR) thresholds of 8- and 16-week-old wild-type (WT), Dusp1 knock-out (KO) and NAC-treated Dusp1 KO mice. Data are presented as mean ± SEM of at least 15 mice per condition. Statistical significance between genotypes was analyzed by one-way ANOVA: * vs. KO, # vs. WT (*, # p < 0.05; **, ## p < 0.01; *** p < 0.001). (B) Representative basal, middle and apical turns hematoxylin-eosin stained cochlear mid-modiolar microphotographs of the organ of Corti (OC) (a–c, g–i and m–o) and spiral ganglion (SG) (d–f, j–l and p–r). SG close-ups (d’–f’, j’–l’ and p’–r’). Scale bars: 25 μm in a, 50 μm in d and 25 μm in d’. Asterisks and arrowheads indicate the absence or presence, respectively, of hair cells and fibers. (C) Representative confocal images of TUNEL-stained (green) basal, middle and apical basilar membrane regions of 16-week-old WT, Dusp1 KO and NAC-treated Dusp1 KO mice co-immunolabeled with Myo7a (red). Asterisks and arrowheads indicate hair cell (HC) loss and TUNEL+ nuclei, respectively. Scale bar: 50 μm. (D) Inner (IHC) and outer (OHC) hair cell number per 200 μm of basal, middle or apical basilar membrane regions. (E) Percentage of TUNEL+ OHCs in 200 μm of basal, middle or apical basilar membrane sections. Data are presented as mean ± SEM of 3 mice per condition. Statistical significance between genotypes was analyzed by one-way ANOVA: * vs. KO, # vs. WT (*, # p < 0.05; **, ## p < 0.01; *** p < 0.001).