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. 2022 Nov 11;14(3):180–201. doi: 10.1093/procel/pwac058

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©The Author(s) 2022. Published by Oxford University Press on behalf of Higher Education Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — Aging-related phenotypes of mouse cochlea. (A) Diagram showing the procedure of aging phenotypical analysis, single cell RNA sequencing (scRNA-seq), and subsequent verification of molecular mechanism. Month, M; Scala Vestibuli, SV; Scala Media, SM; Scala Tympani, ST. (B and C) Line charts showing ABR (B) and DPOAE (C) thresholds of 1-, 2-, 5-, 11-, and 14-month-old mice in response to different frequencies. The ABR or DPAOE thresholds of 2-, 5-, 11-, and 14-month-old mice were compared with those of 1-month-old mice. The quantitative data are presented as the mean ± SEMs (n = 10 mice). Two-tailed Student’s t-test P-values were indicated. Month, M. (D) A recording of the endocochlear potential of 1, 5, and 14 months old mice (1 M, n = 8 mice; 5 M, n = 10 mice; 14 M, n = 8 mice). Data were shown as the mean ± SEMs. Two-tailed Student’s t-test P-values were indicated. Month, M. (E) H&E-staining of outer hair cells in the apical, middle, and basal turns of cochleae from 1-, 5-, and 15-month-old mice. Scale bars, 40 and 10 μm (zoomed-in images). The cell number was counted and quantified. The number of cells is quantified as fold changes relative to that of apical turn in 1-month-old cochlea (n = 5 mice). Two-tailed Student’s t-test P-values were indicated. Month, M. (F) H&E staining of modiolus in the apical, middle, and basal turns from 1-, 5-, and 15-month-old mouse cochleae. Scale bars, 40 μm. The cell density is quantified as fold changes relative to that of apical turn in 1-month-old cochlea (n = 5 mice). Two-tailed Student’s t-test P-values were indicated. (G) H&E staining of stria vascularis in the apical, middle, and basal turns of cochleae from 1-, 5-, and 15-month-old mice. Scale bars, 40 μm and 10 μm (zoomed-in images). The thickness of stria vascularis was counted and quantified as fold changes relative to that of apical turn in 1-month-old cochlea (n = 5 mice). Two-tailed Student’s t-test P-values were indicated. (H) H&E staining of spiral ligament in the apical, middle, and basal turns of cochleae from 1-, 5-, and 15-month-old mice. Scale bars, 80 and 20 μm (zoomed-in images). The apical, middle, and basal spiral ligament cell density were counted and quantified as fold changes relative to that of apical turn in 1-month-old cochlea (n = 5 mice). Two-tailed Student’s t-test P-values were indicated. (I) 4-HNE immunofluorescence staining showed elevated intensity of 4-HNE in 5- or 15-month-old mice compared with that in 1-month-old mice. Scale bars, 40 and 10 μm (zoomed-in images). The relative intensity is quantified as fold changes relative to that of apical turn in 1-month-old cochlea (n = 5 mice). Two-tailed Student’s t-test P-values were indicated.