Alpha-latrotoxin Triggers Extracellular Ca^2＋-dependent Exocytosis and Sensitizes Fusion Machinery in Endocrine Cells

α-Latrotoxin from the venom of black widow spider induces and augments neurotransmitterand hormone release by way of extracellular Ca~(2 ) influx and cellular signal transduction pathways.By usingwhole cell current and capacitance recording,the photolysis of card Ca~(2 ),and Ca~(2 ) microfluorometry andamperometry,we investigated the stimulating effect and mechá(?)ism of α-latrotoxin on exocytosis in ratpancreatic β cells,LβT2 cells and latrophilin plasmid-transfected INS-1 cells.Our data indicated that:(1)α-latrotoxin increased cytosolic Ca~(2 ) concentration through the formation of cation-permitting pores and sub-sequent Ca~(2 ) influx with the presence of extracellular Ca~(2 );(2)α-latrotoxin stimulated exocytosis in normalbath solution and its stimulating effect on secretion was eradicated in Ca~(2 )-free bath solution; and (3)α-latrotoxin sensitized the molecular machinery of fusion through activation of protein kinase C and increasedthe response of cells to Ca~(2 ) photolysed by a flash of ultraviolet light.In summary,α-latrotoxin inducedexocytosis by way of Ca~(2 ) influx and accelerated vesicle fusion by the sensitization of fusion machinery.

Alpha-latrotoxin triggers extracellular Ca(2+)-dependent exocytosis and sensitizes fusion machinery in endocrine cells

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.