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- W64589543 abstract "Regulated secretion in cells is mediated by fusion pore formation between a secretory vesicle and the plasma membrane. The characteristics of the fusion pore are likely to be important in affecting the release of vesicle contents, but have largely remained unknown. Using novel 2-dye fluorescent techniques and 2-photon imaging, we are studying the structure and function of the fusion pore in mouse pancreatic acinar cells. Employing a range of high molecular weight extracellular dyes and recording entry of these dyes in to the vesicles via the open fusion pore we show the fusion pore has a diameter of at least 55 nm. A 2-dye fluorescent technique was then employed to study the dynamics of pore closure. In brief this method consists of stimulating the cells in the presence of an extracellular dye, this dye then enters the lumen of all fused vesicles. One minute after stimulation we then apply a second extracellular dye of a different colour. In most vesicles this second dye enters the vesicle lumen. However, in ~15 % of vesicles, shown to have undergone exocytosis by the presence of the first dye, the second dye does not enter indicating that the fusion pore must have closed. We are currently investigating the functional characteristics of these kinetics by examining the contents of membrane-retained vesicles using an immunohistochemical approach. We are examining the presence of digestive enzymes in populations of vesicles where we have identified fusion pore closure. These studies will provide mechanistic insight into the dynamics and function of the secretory fusion pore in epithelial cells." @default.
- W64589543 created "2016-06-24" @default.
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- W64589543 date "2007-01-01" @default.
- W64589543 modified "2023-09-26" @default.
- W64589543 title "Exocytosis of digestive enzymes: The role of fusion pore dynamics" @default.
- W64589543 hasPublicationYear "2007" @default.
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