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H. Ueno, G. Ferrari, T. Hattori, T. Funaki, D. Saban, S. Chauhan, R. Dana; Nerve Regeneration Associate With Stem Cell Environment. Invest. Ophthalmol. Vis. Sci. 2010;51(13):3753.
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Neurotrophic keratopathy (NK) remains difficult to treat. Impairment of corneal sensory innervation is responsible for corneal epithelial defects, stromal thinning, and perforation. The purpose of this study was to identify the relationship between limbal stem cells and trigeminal innervation using a denervated mouse model.
NK was induced in mice by electro-coagulation of the trigeminal nerve and absence of corneal nerves was assessed by beta-III tubulin immunostaining. We compared the presence of corneal side population cells (SP), which is based on the ability to efflux Hoechst 33342 dye, between normal and denervated corneas by flow cytometry. Real-time polymerase chain reaction was performed to quantify expression of corneal limbal stem cell markers. ATP-binding cassette subfamily G member 2 (ABCG2) was assessed in denervated mice and controls by immunofluorescent microscopic studies.
ABCG2 expression detected with immunostaining was decreased in denervated corneas after 7days. RNA expression levels for both ABCG2 and p63 were significantly decreased in denervated corneas. SP cells from NK mice after 7 days were decreased by approximately 75% when compared with normal corneas.
Corneal stem cells are significantly reduced in this murine model of NK. Additionally, we provide novel evidence for the critical role of innervation in maintaining corneal epithelial cells and/ or the stem cell niche.
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