dependence of corneal stem cells on normal ocular surface innervation

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Dependence of Corneal Stem Cells on Normal Ocular Surface Innervation. H. Ueno, G. Ferrari, T. Hattori, T. Funaki, D.R. Saban , S.K. Chauhan , R. Dana. Schepens Eye Research Institute, Harvard Medical School. The authors have no financial interest in the subject matter of this poster. - PowerPoint PPT Presentation

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Dependence of Corneal Stem Cellson Normal Ocular Surface InnervationH. 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 SchoolBackgroundNeurotrophic keratopathy (NK) is a degenerative disease of the cornea characterized by impaired healing.

Seitz B, Kruse FE.Ophthalmologe. 2005 Corneal sensory nerve degeneration is responsible for corneal epithelial defects, stromal thinning, and perforation.

Purpose

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.

3Methods. Real Time-PCR2. Immunohistochemistry 7days3. Flow cytometry Trigeminal Stereotactic Electrolysis (TSE)no blink reflex

Ophthalmic nerve

G. Ferrari et.al.preparation ABCG2, p63, Hes1ABCG2, p63Side populationCornea4

Contralateral eyeb-tubulin IIIDenervated eyeExpression of beta-III tubulin (neuronal marker) is decreased in the denervated cornea.NormalDenervatedABCG2

p632

DAPIABCG2DAPIDAPIDAPIp63

Expression of stem cell markers (ABCG2 & p63) is decreased in the denervated cornea.Relative Expression of mRNANormal Expression of mRNA:1.0p 0.01p0.01RNA expression levels for both stem cell markers ABCG2 and Hes1 are significantly decreased in denervated corneas. Lgr5 has been suggested to mark of stem cells in multiple adult tissue.Hes1, a major target gene in Notch signaling, regulates the fate and differentiation of various cell types in many developmental systems.Hes1 regulates corneal development and the homeostatic function of corneal epithelial stem/progenitor cells.

7SummaryCorneal stem cells are significantly reduced inthis 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.

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

References1. Touhami A, Grueterich M, Tseng SC. The role of NGF signaling in human limbal epithelium expanded by amniotic membrane Invest Ophthalmol Vis Sci. 2002 ;43:987-94. 2. 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. 2007 ;13:1934-41. 3. 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. 2008 ;86:34-40.

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