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. 2017 Jan 13;16:292–301. doi: 10.1016/j.ebiom.2017.01.004

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© 2017 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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Fig. 4 — Slc4a11^−/− mouse corneal endothelium shows signs of ammonia toxicity and altered glutaminolysis enzymes. (A) Photography of Slc4a11^+/+ and Slc4a11^−/− mouse cornea, shows diffuse edema (increase of light reflection in clear stroma) in 12-week Slc4a11^−/−. (B) H&E staining of 40-week Slc4a11^+/+ and Slc4a11^−/− mouse cornea section shows endothelial vacuolation and Descemet's (basement) membrane thickening. (C) Nitrotyrosine immunostaining shows increased intensity in 40-week old Slc4a11^−/− corneal endothelium, suggesting ammonia toxicity. (D) Quantification of nitrotyrosine staining by mean fluorescence intensity. Quantification of the mean fluorescence intensity was achieved by selecting corneal endothelium layer as region of interest (n = 3 in each group). (E) Real-time qPCR of 12-week Slc4a11^+/+ and Slc4a11^−/− mouse cornea endothelium shows upregulated Gls1 and non-detectable (N.D.) Gls2 in Slc4a11^−/−. (F) Nested-PCR verification of Gls2 and Ggt are not detectable in 12-week Slc4a11^−/− cornea endothelium. (G) Immunostaining verification of Gls1 and Gls2 expression changes in 40-week Slc4a11^+/+ and Slc4a11^−/− mouse cornea endothelium are consistent with real-time qPCR results of 12-week mouse. Data are presented as mean ± s.e.m., *p < 0.05.