CA2-KINASE-C ACTIVATION OF MUCIN GRANULE EXOCYTOSIS IN PERMEABILIZED SPOC1 CELLS( AND PROTEIN)

Mucin secretion by airway goblet cells is under the control of apical P2Y(2), 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) sti mulates 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 Ce ll. Mob. Physiol. 17): L201-L210, 1997]. This apparent independence be tween PKC and Ca2+ in the stimulation of mucin secretion was tested in streptolysin O-permeabilized SPOC1 cells. These cells were fully comp etent to secrete mucin when Ca2+ was elevated from 100 nM to 3.1 mu M for 2 min following permeabilization; the Ca2+ EC50 was 2.29 +/- 0.07 mu M. Permeabilized SPOC1 cells were exposed to PMA or 4 alpha-phorbol at Ca2+ activities ranging from 10 nM to 10 mu M. PMA, but not 4 alph a-phorbol, increased mucin release at all Ca2+ activities tested: at 1 0 nM Ca2+ mucin release was 2.1-fold greater than control and at 4.7 m u M Ca2+ mucin release was maximal (3.6-fold increase). PMA stimulated 27% more mucin release at 4.7 mu M than at 10 nM Ca2+. Hence, SPOC1 c ells possess Ca2+-insensitive, PKC-dependent, and Ca2+-dependent PKC-p otentiated pathways for mucin granule exocytosis.