The Effect of Protein Kinase C Inhibitors on Desensitization of the Murine P2Y2 Receptor

Date of Graduation

Spring 2005

Degree

Master of Science in Cell and Molecular Biology

Department

Biomedical Sciences

Committee Chair

Richard Garrad

Abstract

Cystic Fibrosis (CF) is the most common lethal genetic disease within the Caucasian population. The disease is caused by defects in the cystic fibrosis transmembrane conductance regulator (CFTR) protein that is responsible for chloride ion secretion in airway epithelium. Upon agonist (UTP or ATP) stimulation the G protein-coupled P2Y2 receptor facilities an increase in intracellular calcium that stimulates calcium-dependent chloride anion secretion in airway epithelia. The release of chloride anions via the P2Y2 receptor is an alternative pathway that can bypass the defect present in Cystic Fibrosis. This alternative pathway is short lived due to agonist-induced desensitization. The P2y2 signaling cascade activates protein kinase C (PKC), however, whether PKC plays a specific role in agonist-induced PSY2 receptor cDNA. Spectrofluorometric calcium assays were performed to test the effects of the PKC inhibitors GF109203X, rottlerin, and RO 31-8220, on agonist and PMA-induced PSY2 receptor desensitization. The PKC inhibitor GF109203X inhibited agonist and PMA-induced desensitization, whereas RO 31-8220 inhibited only PMA-induced desensitization. Treatment with the nPKC inhibitor, rottlerin, had no significant effect on agonist or PMA-mediated desensitization. Overall, the results do not support a role for PKC in agonist-induced PSY2 receptor desensitization.

Keywords

P2Y2 receptor, desensitization, protein kinase C, G protein-coupled receptors, cystic fibrosis

Subject Categories

Medical Molecular Biology

Copyright

© Dana K. Tucker

Citation-only

Dissertation/Thesis

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