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. 2017 Aug 14;13(8):e1006969. doi: 10.1371/journal.pgen.1006969

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© 2017 Crompton et al

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Fig 5 — (A) Click-evoked ABR thresholds across a time course show Itga5^tm1Hyn/+; Nisch^edsn/edsn mice exhibit significantly elevated auditory thresholds compared to Itga5^+/+; Nisch^edsn/edsn mice. Additionally, a mild late-onset hearing deficit is observed in Itga5^tm1Hyn/+; Nisch^edsn/+ mice, with onset at 12 wk. Expected ABR threshold range for normal hearing was between 15–30 dB SPL (dashed red lines). Itga5^+/+; Nisch^+/+ n = 14; Itga5^+/+; Nisch^edsn/+ n = 14; Itga5^tm1Hyn/+; Nisch^+/+ n = 15; Itga5^tm1Hyn/+; Nisch^edsn/+ n = 13; Itga5^+/+; Nisch^edsn/edsn n = 8; Itga5^tm1Hyn/+; Nisch^edsn/edsn n = 12. (B-D) Visual inspection of the tympanic membrane was used as a semi-quantitative measure for the prevalence of OM. (B) Itga5^+/+; Nisch^+/+ mice show a very low prevalence of unilateral OM at 4 wk and 6 wk only. (C) Itga5^+/+; Nisch^edsn/edsn mice show a progressive increase in prevalence of OM, whereas (D) Itga5^tm1Hyn/+; Nisch^edsn/edsn mice display a consistently high prevalence of bilateral OM throughout the time course. (E-J) H&E stained transverse sections of the MEC and mucoperiosteum, in 20 wk (E, F) Itga5^+/+ Nisch^+/+, (G, H) Itga5^+/+; Nisch^edsn/edsn and (I, J) Itga5^tm1Hyn/+; Nisch^edsn/edsn mice. Both Itga5^+/+; Nisch^edsn/edsn and Itga5^tm1Hyn/+; Nisch^edsn/edsn mice demonstrate chronic inflammation with an exudate. Inflammation of mucosa was more severe in sections from Itga5^tm1Hyn/+; Nisch^edsn/edsn ears, with increased polypoid exophytic growths and a thick cellular effusion. (K) Blinded assessment of mean mucosal thickness demonstrates significant increases in Itga5^tm1Hyn/+; Nisch^edsn/+, Itga5^+/+; Nisch^edsn/edsn and Itga5^tm1Hyn/+; Nisch^edsn/edsn mice compared to wild-type. Both Itga5^+/+; Nisch^edsn/edsn and Itga5^tm1Hyn/+; Nisch^edsn/edsn mice exhibit significant increases in mucosal thickness compared to Itga5^tm1Hyn/+; Nisch^edsn/+ mice. Itga5^+/+; Nisch^+/+ n = 12; Itga5^+/+; Nisch^edsn/+ n = 8; Itga5^tm1Hyn/+; Nisch^+/+ n = 10; Itga5^tm1Hyn/+; Nisch^edsn/+ n = 18; Itga5^+/+; Nisch^edsn/edsn n = 10; Itga5^tm1Hyn/+; Nisch^edsn/edsn n = 10. (L) To account for the disparities in OM prevalence, the mean mucosal thickness was additionally assessed for OM ears only. Itga5^tm1Hyn/+; Nisch^edsn/edsn OM ears exhibit significant increases in mucosal thickness compared to Itga5^+/+; Nisch^edsn/edsn OM ears. OM only: Itga5^tm1Hyn/+; Nisch^edsn/+ n = 3; Itga5^+/+; Nisch^edsn/edsn n = 8; Itga5^tm1Hyn/+; Nisch^edsn/edsn n = 9. C, cochlea; ET, eustachian tube; E, exudate; MEC, middle ear cavity; MP, mucoperiosteum (arrowheads); TB, temporal bone; TM, tympanic membrane. E, G, I scale bar = 2 mm; F, H, J scale bar = 200 μm. ns P > 0.05; * P < 0.05; ** P < 0.01; *** P < 0.001. Error bars indicate standard error of mean. A Kruskall-Wallis test was performed followed by Dunn’s multiple comparison tests for post-hoc analysis.