Roosvelt University1, Biological, Chemical and Physcial Sciences, Chicago, IL 60605 Roosevelt University2, Biological, Chemical and Physcial Sciences, Chicago, IL 60605
The beta catenin protein serves two main functions in a cell; it physically links E-cadherins and actin filaments, providing the cell its ability to adhere to the extracellular environment, and, via the Wnt pathway, the protein associates with Tcf/Lef transcription factors increasing cellular proliferation. When the Wnt protein binds with the Frizzled receptor on the cell surface, activation of Disheveled suppresses the activity of the axin/APC/GSK-3beta, the regulator of cytoplasmic beta-catenin. Previous studies have indicated a role for the Wnt signaling pathway in the trabecular meshwork (TM), a specialized tissue located in the anterior chamber of the eye proximal to the cornea. The TM is known to play a crucial role in maintaining intraocular pressure through the regulation of the flow of aqueous humor. The TM has been established as an important tissue in glaucoma pathology as a result of its regulation of aqueous humor outflow. In this study, we investigate the role of β‐catenin in bovine TM cells in culture through over expression of the protein. β‐catenin cDNA was amplified from a cDNA library and subcloned into a pTarget mammalian expression vector. The vector was then transfected into the primary bovine TM cell grown from TM tissue explants dissected from fresh bovine eyes. After transfection, a population of bovine TM cells containing the vector were selected and used in further experiments. Over expression of beta-catenin was confirmed through RT-PCR and Western blot analysis. Future studies will include the evaluation of beta-catenin over expression on wound healing and cellular adhesion.