Two phases of zymogen granule lifetime in mouse pancreas: ghost granules linger after exocytosis of contents.
Different cell types show widely divergent mechanisms and kinetics of exocytosis. We investigated these processes in pancreatic acinar cells by using video-rate 2-photon microscopy to image entry of extracellular dye into individual zymogen granules undergoing exocytosis. Fluorescence signals display two distinct phases; an initial peak that then decays over several seconds to a prolonged plateau. Several observations suggest that the first component reflects the binding of dye to the granule contents and their subsequent release into the acinar duct. These observations include: the peak/plateau fluorescence ratio differs between different dyes; the initial fluorescence decay mirrors the loss of granule contents as monitored by differential interference contrast microscopy; and the fall in vesicular fluorescence is accompanied by a rise in fluorescence in the adjacent duct lumen. We thus propose the use of extracellular fluorescent probes as a convenient means to monitor the kinetics of loss of proteinaceous content from secretory granules. In pancreatic acinar cells the fusion pore remains open much longer than required to ensure secretion of the granule contents, and instead the persistent empty 'ghost-granule' may act as a conduit to which secondary granules can fuse and release their contents by compound exocytosis.
Thorn
Peter
P
Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge CB2 1PD, UK. pt207@cam.ac.uk
Parker
Ian
I
D000818:Animals; D002264:Carboxylic Acids; D004792:Enzyme Precursors / Q000378:metabolism; D005089:Exocytosis / Q000502:physiology*; D005456:Fluorescent Dyes; D006834:Hydrazines; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D046790:Pancreas, Exocrine / Q000201:enzymology / Q000502:physiology* / Q000648:ultrastructure*; D012235:Rhodamines; D022142:Secretory Vesicles / Q000502:physiology* / Q000648:ultrastructure*
C417561:Alexa 488 hydrazide; D002264:Carboxylic Acids; D004792:Enzyme Precursors; D005456:Fluorescent Dyes; D006834:Hydrazines; C436864:Oregon Green 488 carboxylic acid; D012235:Rhodamines; C022027:lissamine rhodamine B