Quantitative analysis of exocytosis visualized by a video‐enhanced light/fluorescence microscope reveals two distinct components of exocytosis in RBL‐2H3 cells

FEBS Letters

Suzaki, Etsuko²  Ozawa, Koichiro²  Masujima, Tsutomu²  Kawai, Eriko²  Kobayashi, Hideyuki²  Nonomura, Yoshiaki¹ 

[1]First Department of Pharmacology, Faculty of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113, Japan

[2]Institute of Pharmaceutical Sciences, Hiroshima University School of Medicine, 1-2-3 Kasumi, Minami-ku, Hiroshima 734, Japan

关键词: Exocytosis;    Video-microscope;    Protein kinase C;    Phospholipase D;    RBL-2H3 cell;    RBL-2H3 cells;    rat basophilic leukemia 2H3 cells;    IgE;    immunoglobulin E;    FcϵRI;    high affinity immunoglobulin E receptor;    DNP;    2;    4-dinitrophenol;    DNP-BSA;    antigen consisting of 24 molecules of dinitrophenol conjugated with one molecule of bovine serum albumin;    PIPES;    1;    4-piperazinediethanesulfonic acid;    PI3-kinase;    phosphatidylinositol 3-kinase;    MLCK;    myosin light chain kinase;    PKC;    protein kinase C;    PLD;    phospholipase D;    CCD camera;    charge coupled device camera;    SIT camera;    silicon intensified target camera;    DIC;    differential interference contrast;   

DOI  :  10.1016/S0014-5793(96)01184-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Rat basophilic leukemia (RBL-2H3) cells, which exhibit Ca²⁺-dependent secretion of granules when stimulated with antigen or the Ca²⁺-ionophore A23187, were observed under a video-enhanced light/fluorescence microscope. Exocytotic events of individual granules were visualized in individual cells stimulated with antigen or A23187. Exocytosis of granules stimulated with A23187 showed two peaks in the time course. The earlier one was inhibited by selective inhibitors of protein kinase C (Ro31–8425, Ro31–8220, and chelerythrine) and the other was inhibited by an inhibitor of phosphatidate hydrolase, propranolol. Exocytosis by antigen stimulation, however, showed only one peak, which was inhibited by the selective inhibitors of protein kinase C, but not by propranolol. These results indicate that at least two distinct components of exocytosis exist in RBL2H3 cells.

【 授权许可】

Unknown