Figure 2.

Inoculated P. aeruginosa shows significantly greater adhesion to mouse corneas lacking TRPA1 and TRPV1 nociceptors compared to controls. A. Quantification of bacteria adhering to murine corneas per field of view after inoculation of ~ 1×10¹¹ CFU/mL every hour for 4 h on both intact (no blot) and tissue-paper blotted corneas. TRPA1^−/−/TRPV1^−/− corneas showed significantly greater bacterial adhesion than WT (~ 2.2-fold) with blotting causing a greater difference from WT (~ 3.1-fold). Data are expressed as the mean ± SEM number of bacteria per field of view. * P < 0.05 (Two-way ANOVA). B. Representative images of P. aeruginosa (green) adhering to the murine cornea (red) under each condition. Scale bar = 50 μm. C. Optical slices of the XZ axis showing that bacteria remained surface-associated (green, arrows) and did not penetrate the corneal epithelium (red). Scale bar = 20 μm. D. Quantification of eye wipes over 1 min after capsaicin (100 μM) was dropped onto lightly anesthetized control and RTX-treated mice. RTX-treated mice did not wipe their eyes after capsaicin application versus controls (mean of 34 wipes/min). * P < 0.05 (Mann-Whitney U Test). E. Corneal fluorescein staining observed by slit lamp examination in blotted control eyes compared to the absence of staining in intact (no blot) control or RTX-treated corneas indicating normal epithelial integrity. F. Quantification of bacteria adhering to corneas of RTX-treated mice versus WT controls after 4 h of P. aeruginosa inoculation. Intact corneas (no blot) of RTX-treated mice showed significantly greater adhesion than WT (~ 3.7-fold). After blotting, a greater difference was observed between RTX-treated mice and WT (~ 4.7-fold). Data are expressed as the mean ± SEM number of bacteria per field of view. * P < 0.05 (Two-way ANOVA). G. Representative images of P. aeruginosa (green) adhering to the murine cornea (red) under each condition. Scale bar = 50 μm.