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M.G. Tomich, T.A. Nguyen, A. Jewell, L.J. Takemoto, D.J. Takemoto; Stress-Induced Alterations in Lens Epithelial Cells: Possible Regulatory Role of Gap Junctions . Invest. Ophthalmol. Vis. Sci. 2003;44(13):326.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Earlier studies have shown that oxidative stress alters gap junction activity and communication in cells. Gap junction activity is regulated by a number of factors including drugs, growth factors, and protein kinases. The purpose of this study is to determine how PKC γ regulates the effect of H2O2 stress on lens epithelial cell gap junctions. Methods: DAG, Calphostin C, TPA, PKC γ and α pseudosubstrate inhibitors, LEDGF, EGF, IGF-1 and H2O2 were incubated with whole Sprague-Dawley rat lenses after which dye transfer was determined using lucifer yellow and rhodamine-dextran. The resulting gap junction activity was measured by confocal microscopy. Rabbit lens epithelial N/N1003A cells were grown in tissue culture until 90% confluent. The effect of H2O2 on cell viability was determined by trypan blue dye exclusion. Apoptosis was determined using flow cytometry. Cell culture gap junction activity was determined using the scrape load/dye transfer technique. Results: In lens epithelial cells 120µM H2O2 increased gap junction activity at 15 minutes without cell death. Gap junction activity was normalized by 10 ng/ml LEDGF but IGF-1 had no effect. Conclusions: H2O2 stress induced changes in gap junction activity. This could be controlled by stress-related growth factors such as LEDGF. Data suggest a role for gap junctions in lens epithelial cell stress responses.
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