A novel mouse model for neurotrophic keratopathy: trigeminal nerve stereotactic electrolysis through the brain.
OBJECTIVE
To develop a mouse model of neurotrophic keratopathy by approaching the trigeminal nerve through the brain and to evaluate changes in corneal cell apoptosis and proliferation.


METHODS
Six- to 8-week-old male C57BL/6 mice underwent trigeminal stereotactic electrolysis (TSE) to destroy the ophthalmic branch of the trigeminal nerve. Clinical follow-up using biomicroscopy of the cornea was performed at days 2, 4, 5, and 7. To confirm the effectiveness of the procedure, we examined the gross nerve pathology, blink reflex, and immunohistochemistry of the corneal nerves. TUNEL-positive apoptotic and Ki-67-positive proliferating corneal cells were evaluated to detect changes from the contralateral normal eye.


RESULTS
TSE was confirmed by gross histology of the trigeminal nerve and was considered effective if the corneal blink reflex was completely abolished. TSE totally abolished the blink reflex in 70% of mice and significantly reduced it in the remaining 30%. Animals with absent blink reflex were used for subsequent experiments. In these mice, a progressive corneal degeneration developed, with thinning of the corneal epithelium and eventually perforation after 7 days. In all mice, 48 hours after TSE, corneal nerves were not recognizable histologically. Seven days after TSE, an increase in cellular apoptosis in all the corneal layers and a reduction in proliferation in basal epithelial cells were detected consistently in all mice.


CONCLUSIONS
TSE was able, in most cases, to induce a disease state that reflected clinical neurotrophic keratitis without damaging the periocular structures. Moreover, corneal denervation led to increased apoptosis and reduced proliferation of epithelial cells, formally implicating intact nerve function in regulating epithelial survival and turnover.
Schepens Eye Research Institute, Boston, MA 02114, USA.
Giulio
G
Ferrari
Sunil K
SK
Chauhan
Hiroki
H
Ueno
Nambi
N
Nallasamy
Stefano
S
Gandolfi
Lawrence
L
Borges
Reza
R
Dana
D000818:Animals; D017209:Apoptosis; D001767:Blinking; D001921:Brain / Q000601:surgery; D003315:Cornea / Q000294:innervation* / Q000378:metabolism / Q000473:pathology; D003316:Corneal Diseases / Q000209:etiology* / Q000378:metabolism / Q000473:pathology; D003714:Denervation / Q000379:methods*; D004195:Disease Models, Animal; D004564:Electrocoagulation / Q000379:methods*; D020287:In Situ Nick-End Labeling; D019394:Ki-67 Antigen / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009882:Ophthalmic Nerve / Q000378:metabolism / Q000473:pathology / Q000601:surgery*; D013238:Stereotaxic Techniques
D019394:Ki-67 Antigen