Publication Date
2011
Document Type
Thesis
Committee Members
Norma Adragna (Committee Member), Thomas Brown (Committee Member), David Cool (Committee Member), Peter Lauf (Advisor)
Degree Name
Master of Science (MS)
Abstract
Protein kinase inhibition by staurosporine causes apoptotic volume decrease involving potassium (K) channels in immortalized human B3 lens epithelial cells (LECs). Here, the effect of two pro-apoptotic protein kinase C (PKC) inhibitors [12-O-tetradecanoyl-phorbol-13-acetate (TPA) and chelerythrine (CET)] were studied on membrane K transport in a fetal human LEC line (FHL124) by western blotting, immunofluorescence, ion flux, ATP, apoptosis, and biotinylation assays. Long term TPA exposure (0-6 h) inhibited 75% of Na-K-2Cl cotransport (NKCC). In contrast, short term (0-20 min) exposure to 50 μM CET reduced Na/K pump and NKCC by >90% and >70%, respectively, without retrieval from the membrane into the cytosol, loss of ATP and early signs of apoptosis. CET (10-30 μM) decreased cell K by 33%. Results suggest PKC modulation through the use of pro-apoptotic agents caused major early membrane changes independently affecting K channels, the Na/K pump and NKCC function, prior to rise of any significant apoptosis.
Page Count
128
Department or Program
Department of Pharmacology and Toxicology
Year Degree Awarded
2011
Copyright
Copyright 2011, all rights reserved. This open access ETD is published by Wright State University and OhioLINK.
