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X. Liu, L. Guo, J. Park, M.S. Filla, N. Wang, P.L. Kaufman; C3 Transferase Disrupts CLAN Formation in DEX Treated HTM Cells, Relevant to Glaucoma Gene Therapy . Invest. Ophthalmol. Vis. Sci. 2006;47(13):1864.
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© ARVO (1962-2015); The Authors (2016-present)
To determine the effects of adenovirus–delivered exoenzyme C3 transferase (C3) gene expression on normal or dexamethasone (DEX)–treated cultured human trabecular meshwork (HTM) cells.
DEX–induced changes in actin organization of cultured HTM cells were evaluated. An adenoviral (Ad) vector expressing both C3 and green fluorescent protein (GFP) was constructed and used to transduce cultured HTM cells with or without DEX treatment. Changes in the organization of actin, vinculin and beta–catenin were assessed using immunofluorescence. An Ad vector expressing GFP (AdGFP) only was the control.
Compared to AdGFP–transduced cells, AdC3GFP–transduced cells demonstrated disrupted actin cytoskeleton, and reduced vinculin and beta–catenin staining. DEX induced formation of cross linked actin networks (CLANs) in cultured HTM cells, and, when the cells were treated with AdC3GFP, CLAN structure was disrupted.
C3 transduction is effective in disrupting the actin cytoskeleton, cellular adhesions, and DEX–induced CLANs, in HTM cells, providing molecular and cellular bases for earlier reports of the outflow facility increasing effects of C3 expression in monkey organ cultured anterior segment.
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