Quantitative analysis of exocytosis visualized by a video-enhanced light/fluorescence microscope reveals two distinct components of exocytosis in RBL-2H3 cells.
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 RBL-2H3 cells.
Ozawa
K
K
Institute of Pharmaceutical Sciences, Hiroshima University School of Medicine, Japan.
Kobayashi
H
H
Kawai
E
E
Suzaki
E
E
Nonomura
Y
Y
Masujima
T
T
D000818:Animals; D000941:Antigens / Q000494:pharmacology; D000001:Calcimycin / Q000494:pharmacology; D004140:Dinitrophenols / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005089:Exocytosis / Q000187:drug effects / Q000502:physiology*; D006632:Histamine / Q000378:metabolism; D007089:Image Enhancement / Q000379:methods; D007211:Indoles / Q000494:pharmacology; D015471:Leukemia, Basophilic, Acute / Q000378:metabolism* / Q000473:pathology; D008301:Maleimides / Q000494:pharmacology; D008856:Microscopy, Fluorescence / Q000379:methods*; D018715:Microscopy, Video / Q000379:methods*; D009281:Naphthalenes / Q000494:pharmacology; D010739:Phospholipase D / Q000378:metabolism; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D051381:Rats; D012701:Serotonin / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D013997:Time Factors; D014407:Tumor Cells, Cultured
D000941:Antigens; D004140:Dinitrophenols; D004791:Enzyme Inhibitors; D007211:Indoles; D008301:Maleimides; D009281:Naphthalenes; C061465:calphostin complex; C034638:dinitrophenyl-bovine serum albumin; C071924:Ro 31-8425; D012710:Serum Albumin, Bovine; D012701:Serotonin; D000001:Calcimycin; D006632:Histamine; D011493:Protein Kinase C; D010739:Phospholipase D; C064758:Ro 31-8220