title: Dynamic imaging of single secretory granule in cultured PC12 cells.
abstract: Fluorescent labeling and dynamic imaging of secretory vesicles are new powerful means to study the mechanisms of protein and membrane trafficking. The large dense-core granules in PC12 cells were labeled by transfecting EGFP-hpNPY chimera and imaged with epi-fluorescence and evanescent field (EF) fluorescence simultaneously. Under epi-fluorescence illumination, the cells exhibited nearly uniform fluorescence, however, EF-fluorescence excitation revealed distinct fluorescent spots corresponding to GFP-labeled granules. The trafficking, docking and fusing with plasma membrane of individual granule in cultured PC12 cells were observed directly by EF fluorescence imaging.
authors.affiliation: Institute of Biophysics and Biochemistry, Huazhong University of Science and Technology, Wuhan 430074, China. txu@hust.edu.cn
authors.forename: Zheng-Xing
authors.initials: ZX
authors.lastname: Wu
authors.forename: Sheng
authors.initials: S
authors.lastname: Xia
authors.forename: Liang
authors.initials: L
authors.lastname: Xu
authors.forename: Li
authors.initials: L
authors.lastname: Bai
authors.forename: Tao
authors.initials: T
authors.lastname: Xu
mesh_terms: D000818:Animals; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods*; D016716:PC12 Cells / Q000378:metabolism*; D021381:Protein Transport; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D022142:Secretory Vesicles / Q000378:metabolism*; D014162:Transfection
chemical_list: D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins