Fig. 7: Local substance P blockade ameliorates contralateral corneal nerve dysfunction associated with unilateral dry eye. | Experimental & Molecular Medicine

Fig. 7: Local substance P blockade ameliorates contralateral corneal nerve dysfunction associated with unilateral dry eye.

From: A transient receptor potential vanilloid 1-dependent corneal–trigeminal neuroinflammatory circuit promotes corneal neuropathy

Fig. 7

a Unilateral DED was surgically induced in WT mice of both sexes by excising the right extraorbital lacrimal gland only and and either 10 mg/ml fosaprepitant (SPbl), a neurokinin-1 receptor antagonist or saline (PBS) was applied four times per day for 10 days on the contralateral (Contra) eye starting immediately after the surgery. b Cumulative data (left) and representative micrographs (right) of corneal dextran-fluorescein uptake in the Contra eyes of PBS and SP blocker-treated mice. Data are shown as the MFI calculated with ImageJ software (Materials and methods). c The number (left) of proliferating (Ki67+, green) cells within the epithelial basal layer and representative micrographs (right) of corneal wholemounts obtained 10 days after DED induction. E-cadherin (epithelial-specific) stained in red. d, e The corneal mechanosensitivity (d) and ocular capsaicin sensitivity (e) in Contra eyes of PBS and SP blocker-treated mice on days 0, 5 and 10 of unilateral DED induction shown as individual data points (left) and overall progression curves (right). f The eye-closing ratio in Contra eyes of PBS and SP blocker-treated mice on day 10 of unilateral DED induction. g–j The quantification of intraepithelial corneal innervation at three different levels by anti-tubulin β3 staining of corneal wholemounts of the PBS- and SP blocker-treated Contra eyes after 10 days of DED induction: subapical (g) (percent area occupied by nerve endings) and midepithelial (h) (count of nerve endings per field) nerve ending density and complexity of subbasal nerves (i) (sum of intersections at all Sholl radii) and representative micrographs (j). In b–h the black and red dotted lines correspond to sham-operated mice and ipsilateral eyes of unilateral DED eyes, respectively. To compare means, two-way ANOVA was used for b c and e (time and treatment) with Dunnett’s post hoc test, and Student’s t-test was applied in c and f–i. *P < 0.05, **P < 0.01. ns, not significant.

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