title: The plasma membrane Q-SNARE syntaxin 2 enters the zymogen granule membrane during exocytosis in the pancreatic acinar cell.
abstract: During exocytosis in the pancreatic acinar cell, zymogen granules fuse directly with the apical plasma membrane and also with granules that have themselves fused with the plasma membrane. Together, these primary and secondary fusion events constitute the process of compound exocytosis. It has been suggested that the sequential nature of primary and secondary fusion is a consequence of the requirement for plasma membrane soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptors, such as syntaxin 2, to enter the membrane of the primary fused granule. We have tested this possibility by determining the location of syntaxin 2 in unstimulated and stimulated pancreatic acini. Syntaxin 2 was imaged by confocal immunofluorescence microscopy. Fused granules were detected both through their filling with the aqueous dye lysine-fixable Texas Red-dextran and through the decoration of their cytoplasmic surfaces with filamentous actin. In unstimulated cells, syntaxin 2 was exclusively present on the apical plasma membrane. In contrast, after stimulation, syntaxin 2 had moved into the membranes of fused granules, as judged by its location around dye-filled structures of 1-mum diameter that were coated with filamentous actin. At long times of stimulation (5 min), the majority (85%) of dye-filled granules were also positive for syntaxin 2. In contrast, at shorter times (1 min), more dye-filled granules (29%) were syntaxin 2-negative. We conclude that syntaxin 2 enters the membrane of a fused zymogen granule after the opening of the fusion pore, and we suggest that this movement might permit the onset of secondary fusion.
authors.affiliation: Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge CB2 1PD, United Kingdom.
authors.forename: James A
authors.initials: JA
authors.lastname: Pickett
authors.forename: Peter
authors.initials: P
authors.lastname: Thorn
authors.forename: J Michael
authors.initials: JM
authors.lastname: Edwardson
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D000954:Antigens, Surface / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism*; D016764:Cell Polarity; D003911:Dextrans; D004792:Enzyme Precursors / Q000378:metabolism*; D005089:Exocytosis; D007425:Intracellular Membranes / Q000378:metabolism*; D051379:Mice; D008856:Microscopy, Fluorescence; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D010179:Pancreas / Q000166:cytology* / Q000187:drug effects; D051381:Rats; D050600:SNARE Proteins; D022142:Secretory Vesicles / Q000378:metabolism*; D050827:Syntaxin 1; D033921:Vesicular Transport Proteins / Q000378:metabolism; D014966:Xanthenes
chemical_list: D000199:Actins; D000954:Antigens, Surface; D003911:Dextrans; D004792:Enzyme Precursors; D009419:Nerve Tissue Proteins; D050600:SNARE Proteins; D050827:Syntaxin 1; D033921:Vesicular Transport Proteins; D014966:Xanthenes; C034657:Texas red