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James Spurlin, Peter Lwigale; Innervation of embryonic corneas during wound healing. Invest. Ophthalmol. Vis. Sci. 2013;54(15):3901.
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© ARVO (1962-2015); The Authors (2016-present)
Determine if embryonic corneal wounds are innervated during regeneration.
We created a single wound traversing the epithelium and anterior stroma in the central cornea of one eye in each embryo at E7, the other eye of the same embryo serves as a negative control. Wounded and unwounded stage matched corneas were collected between E7 and E18 and analyzed for innervation following immunostaining with beta-neurotubulin antibody (TUJ1). Temporal expression of the axon guidance molecule, Semaphorin3A, was evaluated in wounded corneas using qPCR.
During regeneration of embryonic corneas, nerves do not project into the non-healed regions of the cornea but innervate sheet of epithelial cells that migrate into the wound. The rate of innervation of wounded embryonic corneas appears to follow the rate of re-epithelialization. Interestingly, Sema3A transcript levels are elevated in wounded embryonic corneas, peaking at 1.7 fold increase by 3 days post wound (dpw), prior to the onset of wound re-epithelialization. Sema3A transcript levels in wounded corneas decrease by 5 dpw and approach basal transcript levels detected in paired control corneas by 7dpw. By 11dpw, there is no difference in nerve density and innervation patterns between regenerated corneas and stage matched non-wounded controls.
These data suggest that axon projections into the wounded embryonic cornea are initially inhibited at the wound periphery by the increased expression of Sema3A. However, the high level of Sema3A is transient, which permits appropriate innervation of the regenerated corneas.
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