BrdU标记

A Mouse Model for the Studyof Recurrent CornealEpithelial Erosions: ␣9␤1 Integrin Implicated inProgression of the DiseaseSonali Pal-Ghosh,1 Ahdeah Pajoohesh-Ganji,1,2 Marcus Brown,1 and MaryAnn Stepp1,3PURPOSE. To describe an in vivo mouse model for the study ofrecurrent corneal erosion syndrome (RCES) in mice and tocharacterize the changes in ␣9 integrin expression duringwound healing.METHODS. Corneal epithelial debridement wounds of two sizes(1.5 and 2.5 mm) were made on the ocular surface of BALB/cmice and were evaluated at various times after wounding.Corneas were processed either as whole mounts and stainedwith propidium iodide and an antibody against ␣9 integrin orfor bromodeoxyuridine analyses of cell proliferation. A sepa-rate study involved analyses of corneal wound healing overtime in individual mice with large and small debridementwounds. Mice were anesthetized once per week and theircorneas stained with fluorescein to assess the quality of thecorneal epithelium. After 6 weeks, mice were killed and eyesprocessed for study by immunofluorescence in either wholemounts or frozen sections.RESULTS. Whole mount confocal microscopy showed openwounds on the ocular surface of mice at 1 and 2 weeks afterlarge wounds were created, but not after small wounds. Inaddition, ␣9 integrin was upregulated during healing, andchanges were observed in ␣9 integrin localization at the limbuswith large wounds but not with small wounds. Although only1 of 16 corneas with small wounds had erosions at 1 and 2weeks, 11 of 16 with large wounds had erosions. However, by6 weeks, 13 of 16 eyes showed signs of erosion whetherwounds were small or large. With large wounds, RCES corneasfrequently showed numerous goblet cells adjacent to ...