Abstract
Abstract: :
Objective: To determine the expression profiles of α3, α6, α9, ß1 and ß4 integrins on the mouse ocular surface using whole mounts. Methods: Eyes of adult BalbC mice were sutured on the left temporal region to maintain orientation and labeled as left or right eyes. They were then fixed in a mixture of methanol and dimethyl sulfoxide, and stored in 100% methanol. Prior to staining, the lens and posterior tissues were discarded and the anterior segments were used for these experiments. For α9 integrin, eyes where gradually transferred to PBS, incubated with primary antibody overnight, and incubated with secondary antibody for 2 hours. Eyes were flattened on black filters, and coverslipped, For other integrins, the eyes were incubated with 30% triton X–100 in PBS for 5 minutes without going through the gradual series of methanol. They were then washed with PBS and the same procedure was followed. The images were captured using a confocal microscope. Results: Integrins are differentially expressed on the mouse ocular surface in the central cornea versus the limbus, where the stem cells reside. α9 integrin is expressed at the limbal region exclusively, whereas, preliminary data on α3, α6, ß1 and ß4 integrins show that these integrins are present on the entire ocular surface including the limbus and the conjunctiva. Conclusions:Integrins have unique distributions on the mouse ocular surface suggesting that α3ß1, α9ß1, and α6ß4 play distinct roles in maintaining the ocular surface epithelia. These findings are currentlybeing used to study the properties of label–retaining cells at the limbus. Research supported by EY13559–03.
Keywords: cornea: epithelium • cell adhesions/cell junctions • microscopy: light/fluorescence/immunohistochemistry