title: Spatial distribution and temporal evolution of DRONPA-fused SNAP25 clusters in adrenal chromaffin cells.
abstract: Sub-diffraction imaging of plasma membrane localized proteins, such as the SNARE (Soluble NSF Attachment Protein Receptor) proteins involved in exocytosis, in fixed cells have resulted in images with high spatial resolution, at the expense of dynamical information. Here, we have imaged localized fluorescence bursts of DRONPA-fused SNAP-25 molecules in live chromaffin cells by Total Internal Reflection Fluorescence (TIRF) imaging. We find that this method allows tracking protein cluster dynamics over relatively long times (∼20 min.), partly due to the diffusion into the TIRF field of fresh molecules, making possible the simultaneous identification of cluster size, location and temporal evolution. The results indicate that the DRONPA-fused SNAP-25 clusters display rich dynamics, going from staying constant to disappearing and reappearing in specific cluster domains within minutes.
authors.affiliation: Department of Neuroscience and Pharmacology and Center for Biomembranes in Nanomedicine (CBN), University of Copenhagen, Blegdamsvej 3, 2200 Copenhagen, Denmark. jakobbs@sund.ku.dk.
authors.forename: Yasuko
authors.initials: Y
authors.lastname: Antoku
authors.forename: Peter
authors.initials: P
authors.lastname: Dedecker
authors.forename: Paulo S
authors.initials: PS
authors.lastname: Pinheiro
authors.forename: Tom
authors.initials: T
authors.lastname: Vosch
authors.forename: Jakob Balslev
authors.initials: JB
authors.lastname: Sørensen
mesh_terms: D000311:Adrenal Glands / Q000378:metabolism*; D000818:Animals; D019439:Chromaffin Cells / Q000378:metabolism*; D004058:Diffusion; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D018345:Mice, Knockout; D008856:Microscopy, Fluorescence; D050825:Synaptosomal-Associated Protein 25 / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors
chemical_list: D008164:Luminescent Proteins; C494209:Snap25 protein, mouse; D050825:Synaptosomal-Associated Protein 25