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.

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Dependence of Corneal Stem Cells on Normal Ocular Surface Innervation

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Dependence of corneal stem cells on normal ocular surface innervation

Dependence of Corneal Stem Cellson 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.


Dependence of corneal stem cells on normal ocular surface innervation

Background

Neurotrophickeratopathy (NK) is a degenerative disease of the cornea characterized by impaired healing.

Corneal sensory nerve degeneration is responsible for corneal epithelial defects, stromal thinning, and perforation.

Seitz B, Kruse FE.

Ophthalmologe. 2005


Purpose

Purpose

  • To identify the relationship between limbal stem cells and trigeminal innervation using a mouse model of corneal denervation.

  • Hypothesis: Neurotrophickeratopathy results in epithelial stem cell deficiency.

  • Approach: Analysis of stem cells in a mouse model of neurotrophickeratopathy.


Dependence of corneal stem cells on normal ocular surface innervation

Methods

Trigeminal Stereotactic Electrolysis (TSE)

no blink reflex

7days

Cornea

Ophthalmic nerve

1. Real Time-PCR

ABCG2, p63, Hes1

2. Immunohistochemistry

ABCG2, p63

3. Flow cytometry

G. Ferrari et.al.

preparation

Side population


Dependence of corneal stem cells on normal ocular surface innervation

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

Contralateral eye

Denervated eye

b-tubulinIII


Dependence of corneal stem cells on normal ocular surface innervation

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

Denervated

Normal

2

ABCG2

DAPI

ABCG2

DAPI

p63

DAPI

p63

DAPI


Dependence of corneal stem cells on normal ocular surface innervation

RNA expression levels for both stem cell markers ABCG2 and Hes1 are significantly decreased in denervated corneas.

Relative Expression of mRNA

Normal Expression

of mRNA:1.0

p≦0.01

p≦ 0.01


Summary

Summary

Corneal stem cells are significantly reduced in

this murine model of neurotrophickeratopathy.

Expression of stem cell markers (ABCG2, P63, Hes1) in

denervated corneas is decreased at both the mRNA and

protein levels.


Conclusion

Conclusion

Our novel data clearly demonstrate that

corneal nerves play a critical role in maintaining

the corneal stem cells.


References

References

1. Touhami A, Grueterich M, Tseng SC. The role of NGF

signalingin 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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