From: Sonic Hedgehog Expression and Role in Healing Corneal Epithelium

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From: Sonic Hedgehog Expression and Role in Healing Corneal Epithelium Invest. Ophthalmol. Vis. Sci.. 2004;45(8):2577-2585. doi:10.1167/iovs.04-0001 Figure Legend: Activation of Gli-3 signaling and proliferation of epithelial cells in healing epithelium postdebridement in vivo. (a) Immunofluorescent detection of Gli-3 in healing rat corneal and limbal epithelium. Gli-3 protein is observed in the cell cytoplasm of normal, uninjured, corneal (aA) and limbal (aB) epithelia. At 2 hours postdebridement, nuclei of healing corneal epithelium remain negative for Gli-3 (aC), whereas nuclear translocation of Gli-3 was detected in limbal epithelium (arrows, aD). Gli-3 protein is then observed in many cell nuclei of migrating epithelium at 6 hours (arrows, aE) and 12 hours (data not shown) postinjury. At these timepoints only a few limbal basal cells show a nuclear Gli-3 immunoreactivity (arrow, aF). At 24 hours postinjury, Gli-3 immunofluorescent staining is no longer observed in both corneal (aG) and limbal (aH) epithelia. Inset in (aD): a high magnification picture of nuclear localization of Gli-3. Bar, 100 μm. (b) Distribution of Ki67-positive cells in corneal epithelium at intervals of healing postdebridement. Many Ki67-labeled cells are observed in the basal layer of uninjured corneal epithelium (bA, bB). The number of Ki67-positive cells decreases in healing epithelia at 1 hour (not illustrated), 2 hours (bC, bD), and 6 hours (bE, bF) postdebridement. At 12 hours postinjury, the healing epithelium is still less proliferative, whereas many of the repopulated keratocytes are labeled with anti-Ki67 antibody (not illustrated). At 24 hours, nuclei of many cells of central epithelium resurfacing the defect are labeled with anti-Ki67 antibody (bG, bH). Immunofluorescent staining (bB, bD, bF, and bH: DAPI nuclear staining). (c) Histogram of Ki67-positive epithelial cells at each healing interval. Bar, 100 μm. Date of download: 1/3/2018 The Association for Research in Vision and Ophthalmology Copyright © 2018. All rights reserved.