P C Lu1, H Ye, M Maeda, D T Azar. 1. Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, and Department of Ophthalmology, Harvard Medical School, Boston 02114, USA.
Abstract
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.
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.
Authors: John P T Higgins; Lingli Wang; Neeraja Kambham; Kelli Montgomery; Veronica Mason; Stefanie U Vogelmann; Kevin V Lemley; Patrick O Brown; James D Brooks; Matt van de Rijn Journal: Mol Biol Cell Date: 2003-12-02 Impact factor: 4.138
Authors: M Saghizadeh; D J Brown; R Castellon; M Chwa; G H Huang; J Y Ljubimova; S Rosenberg; K S Spirin; R B Stolitenko; W Adachi; S Kinoshita; G Murphy; L J Windsor; M C Kenney; A V Ljubimov Journal: Am J Pathol Date: 2001-02 Impact factor: 4.307