Abstract
alpha-Latrotoxin from the venom of black widow spider induces and augments neurotransmitter and hormone release by way of extracellular Ca(2+) influx and cellular signal transduction pathways. By using whole cell current and capacitance recording, the photolysis of caged Ca(2+), and Ca(2+) microfluorometry and amperometry, we investigated the stimulating effect and mechanism of alpha-latrotoxin on exocytosis in rat pancreatic beta cells, LbetaT2 cells and latrophilin plasmid-transfected INS-1 cells. Our data indicated that: (1) alpha-latrotoxin increased cytosolic Ca(2+) concentration through the formation of cation-permitting pores and subsequent Ca(2+) influx with the presence of extracellular Ca(2+); (2) alpha-latrotoxin stimulated exocytosis in normal bath solution and its stimulating effect on secretion was eradicated in Ca(2+)-free bath solution; and (3) alpha-latrotoxin sensitized the molecular machinery of fusion through activation of protein kinase C and increased the response of cells to Ca(2+) photolyzed by a flash of ultraviolet light. In summary, alpha-latrotoxin induced exocytosis by way of Ca(2+) influx and accelerated vesicle fusion by the sensitization of fusion machinery.
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