Barium and Calcium Stimulate Secretion from Digitonin-Permeabilized Bovine Adrenal Chromaffin Cells by Similar Pathways

We compared the characteristics of secretion stimulated by EGTA-buffered Ba 2+ - and Ca 2+ -containing solutions in digitonin-permeabilized bovine adrenal chromaffin cells. Half-maximal secretion occurred at approximately 100 Μ M Ba 2+ or 1 Μ M Ca 2+ . Ba 2+ -stimulated release was not due to release of sequestered intracellular Ca 2+ because at a constant free Ba 2+ concentration, increasing unbound EGTA did not diminish the extent of release due to Ba 2+ . The maximal extents of Ba 2+ - and Ca 2+ -dependent secretion in the absence of MgATP were identical. MgATP enhanced Ba 2+ -induced secretion to a lesser extent than Ca 2+ -induced secretion. Half-maximal concentrations of Ba 2+ and Ca 2+ , when added together to cells, yielded approximately additive amounts of secretion. Maximal concentrations of Ba 2+ and Ca 2+ when added together to cells for 2 or 15 min were not additive. Tetanus toxin inhibited Ba 2+ - and Ca 2+ -dependent secretion to a similar extent. Ba 2+ , unlike Ca 2+ , did not activate polyphosphoinositide-specific phospholipase C. These data indicate that (1) Ba 2+ directly stimulates exocytosis, (2) Ba 2+ -induced secretion is stimulated to a lesser extent than Ca 2+ -dependent secretion by MgATP, (3) Ba 2+ and Ca 2+ use similar pathways to trigger exocytosis, and (4) exocytosis from permeabilized cells does not require activation of polyphosphoinositide-specific phospholipase C.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/66323/1/j.1471-4159.1992.tb09771.x.pd

Regulation of exocytosis by the exocyst subunit Sec6 and the SM protein Sec1

The Sec6 subunit of the multisubunit exocyst tethering complex interacts with the Sec1/Munc18 protein Sec1 and with the t-SNARE Sec9. Assembly of the exocyst upon vesicle arrival at sites of secretion is proposed to release Sec9 for SNARE complex assembly and to recruit Sec1 for interaction with SNARE complexes to facilitate fusion