adenosine-5--o-(3-thiotriphosphate) and phorbol

Mucin secretion by airway goblet cells is under the control of apical P2Y2, phospholipase C-coupled purinergic receptors. In SPOC1 cells, the mobilization of intracellular Ca2+ by ionomycin or the activation of protein kinase C (PKC) by phorbol 12-myristate 13-acetate (PMA) stimulates mucin secretion in a fully additive fashion [L. H. Abdullah, J. D. Conway, J. A. Cohn, and C. W. Davis. Am. J. Physiol. 273 (Lung Cell. Mol. Physiol. 17): L201-L210, 1997]. This apparent independence between PKC and Ca2+ in the stimulation of mucin secretion was tested in streptolysin O-permeabilized SPOC1 cells. These cells were fully competent to secrete mucin when Ca2+ was elevated from 100 nM to 3.1 microM for 2 min following permeabilization; the Ca2+ EC50 was 2.29 +/- 0.07 microM. Permeabilized SPOC1 cells were exposed to PMA or 4alpha-phorbol at Ca2+ activities ranging from 10 nM to 10 microM. PMA, but not 4alpha-phorbol, increased mucin release at all Ca2+ activities tested: at 10 nM Ca2+ mucin release was 2.1-fold greater than control and at 4.7 microM Ca2+ mucin release was maximal (3.6-fold increase). PMA stimulated 27% more mucin release at 4.7 microM than at 10 nM Ca2+. Hence, SPOC1 cells possess Ca2+-insensitive, PKC-dependent, and Ca2+-dependent PKC-potentiated pathways for mucin granule exocytosis.

Topics: Adenosine Triphosphate; Animals; Bacterial Proteins; Calcium; Cell Line; Cell Membrane Permeability; Cytoplasmic Granules; Enzyme Activation; Exocytosis; Kinetics; Mucins; Mucous Membrane; Phorbols; Protein Kinase C; Rats; Streptolysins; Tetradecanoylphorbol Acetate; Trachea; Uridine Triphosphate