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P C Lu, H Ye, M Maeda, D T Azar; Immunolocalization and gene expression of matrilysin during corneal wound healing.. Invest. Ophthalmol. Vis. Sci. 1999;40(1):20-27. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
PURPOSE: To investigate the protein level of matrilysin and stromelysin-1 and the gene expression of matrilysin in rat corneas after excimer keratectomy using immunofluorescence staining, reverse transcriptase-polymerase Chain Reaction (RT-PCR), and in situ hybridization. METHODS: Rat corneas were treated with 3-mm excimer laser keratectomy (193-nm ArF). Unwounded corneas served as controls. Confocal microscopy was used to immunolocalize matrilysin protein at 6 hours, 1 day, 3 days, 1 week, 4 weeks, and 8 weeks after surgery. RT-PCR was performed to analyze matrilysin mRNA in unwounded controls and in 3-day wounded corneas. In situ hybridization was performed to localize matrilysin mRNA. RESULTS: Matrilysin was immunolocalized to the epithelial layers of unwounded and wounded corneas, predominantly to the leading edge at 6 hours and 1 day after wounding and to the epithelial layer at 3 to 7 days after surgery. Stromelysin-1 was expressed in the deep stromal layer in the first 3 days after wounding and in the area of newly synthesized stromal matrix 1 week after surgery. Upregulation of matrilysin expression 3 days after wounding was confirmed by RT-PCR. In situ hybridization revealed that the gene expression of matrilysin in rat corneas was upregulated during the migration phase (6 hours, 1 day) and that matrilysin mRNA was predominantly localized to the basal cell layers during the subsequent cell proliferation phase (7 and 14 days). CONCLUSIONS: Basal epithelial cells express matrilysin during the migration proliferation phase of corneal wound healing after excimer keratectomy.
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