H. Ueno, G. Ferrari, T. Hattori, T. Funaki, D.R. Saban, S.K. Chauhan, R. Dana The authors have no financial interest in the subject matter of this poster.

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H. Ueno, G. Ferrari, T. Hattori, T. Funaki, D.R. Saban, S.K. Chauhan, R. Dana The authors have no financial interest in the subject matter of this poster. Schepens Eye Research Institute, Harvard Medical School

Background Neurotrophic keratopathy (NK) is a degenerative disease of the cornea characterized by impaired healing. Seitz B, Kruse FE. Ophthalmologe Corneal sensory nerve degeneration is responsible for corneal epithelial defects, stromal thinning, and perforation.

To identify the relationship between limbal stem cells and trigeminal innervation using a mouse model of corneal denervation. Hypothesis: Neurotrophic keratopathy results in epithelial stem cell deficiency. Approach: Analysis of stem cells in a mouse model of neurotrophic keratopathy.

Methods １. Real Time-PCR 2. Immunohistochemistry 7days 3. Flow cytometry Trigeminal Stereotactic Electrolysis (TSE) no blink reflex Ophthalmic nerve G. Ferrari et.al. preparation ABCG2, p63, Hes1 ABCG2, p63 Side population Cornea

Contralateral eye  -tubulin III Denervated eye Expression of beta-III tubulin (neuronal marker) is decreased in the denervated cornea.

Normal Denervated ABCG2 p63 2 DAPIABCG2DAPI p63 Expression of stem cell markers (ABCG2 & p63) is decreased in the denervated cornea.

Relative Expression of mRNA Normal Expression of mRNA:1.0 p ≦ 0.01 RNA expression levels for both stem cell markers ABCG2 and Hes1 are significantly decreased in denervated corneas.

Corneal stem cells are significantly reduced in this murine model of neurotrophic keratopathy. Expression of stem cell markers (ABCG2, P63, Hes1) in denervated corneas is decreased at both the mRNA and protein levels.

Our novel data clearly demonstrate that corneal nerves play a critical role in maintaining the corneal stem cells.

1. Touhami A, Grueterich M, Tseng SC. The role of NGF signaling in human limbal epithelium expanded by amniotic membrane Invest Ophthalmol Vis Sci ;43: Qi H, Chuang EY, Yoon KC, de Paiva CS, Shine HD, Jones DB, Pflugfelder SC, Li DQ. Patterned expression of neurotrophic factors and receptors in human limbal and corneal regions. Mol Vis ;13: Qi H, Li DQ, Shine HD, Chen Z, Yoon KC, Jones DB, Pflugfelder SC. Nerve growth factor and its receptor TrkA serve as potential markers for human corneal epithelial progenitor cells. Exp Eye Res ;86:34-40.