Dependence of Corneal Stem Cellson Normal Ocular Surface Innervation
H. Ueno, G. Ferrari, T. Hattori,
T. Funaki, D.R. Saban, S.K. Chauhan,
R. DanaThe 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. 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.
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 eyeb
-tub
ulin
III
Denervated eye
Expression of beta-III tubulin (neuronal marker) is decreased in the denervated cornea.
Normal Denervated
ABCG2
p63
2DAPI ABCG2 DAPI
DAPI DAPIp63
Expression of stem cell markers (ABCG2 & p63) is decreased in the denervated cornea.
ABCG2 p63 Hes1-1.66533453693773E-16
0.2
0.4
0.6
0.8
1
1.2
Rel
ativ
e E
xpre
ssio
n of
mR
NA Normal Expression
of mRNA:1.0
p 0.01≦p 0.01≦
RNA expression levels for both stem cell markers ABCG2 and Hes1 are significantly decreased in denervated corneas.
Summary
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.
Conclusion
Our novel data clearly demonstrate that
corneal nerves play a critical role in maintaining
the corneal stem cells.
References
1. 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.
