Dynamic imaging of single secretory granule in cultured PC12 cells.
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.
Wu
Zheng-Xing
ZX
Institute of Biophysics and Biochemistry, Huazhong University of Science and Technology, Wuhan 430074, China. txu@hust.edu.cn
Xia
Sheng
S
Xu
Liang
L
Bai
Li
L
Xu
Tao
T
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
D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins