PMA and calcium ionophore induce myosin and F-actin rearrangement during histamine secretion from RBL-2H3 cells.
Rat basophilic leukemia (RBL-2H3) cells undergo morphological and cytoskeletal changes during antigen-induced secretion of allergic mediators. The exact role these changes play in the process of secretion is unclear. Using confocal microscopy we now show that PMA+A23187 causes extensive F-actin rearrangements during secretion of [3H] 5-HT. We also describe for the first time the association of myosin with F-actin during this secretory process. In unstimulated cells, myosin and F-actin are concentrated at the plasma membrane with no evidence of stress fibres. Upon addition of PMA or A23187, both F-actin and myosin are rearranged into membrane ruffles and discrete aggregations (foci), followed by the formation of parallel stress fibres located on the ventral membrane. This is in contrast to reports in other cell types in which PMA has been described as causing the disruption of F-actin stress fibres. The time course of secretion coincides with the formation of the foci and ruffles whilst the stress fibres form after the majority of secretion has occurred. These changes are accompanied by a 40% decrease in cell height and a two-fold increase in cell spreading and they occur in the absence of extracellular calcium but are inhibited by the protein kinase C inhibitor, Bisindolylmaleimide, which also inhibits secretion. The formation of myosin-decorated stress fibres, foci, and ruffles is not sufficient to cause secretion, as PMA alone induces these changes without any secretion. The relevance of actin and myosin rearrangements for the regulation of secretion is discussed.
Ludowyke
R I
RI
Centre for Immunology, St. Vincent's Hospital, Sydney, Australia.
Kawasugi
K
K
French
P W
PW
D000199:Actins / Q000032:analysis* / Q000648:ultrastructure; D000818:Animals; D000001:Calcimycin / Q000494:pharmacology*; D002118:Calcium / Q000494:pharmacology; D002462:Cell Membrane; D048429:Cell Size / Q000187:drug effects; D006636:Histamine Release; D015471:Leukemia, Basophilic, Acute; D008301:Maleimides / Q000494:pharmacology; D008407:Mast Cells / Q000737:chemistry / Q000187:drug effects* / Q000648:ultrastructure; D008856:Microscopy, Fluorescence; D009218:Myosins / Q000032:analysis* / Q000648:ultrastructure; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D051381:Rats; D012701:Serotonin / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014316:Tritium; D014404:Tubulin / Q000032:analysis; D014407:Tumor Cells, Cultured; D014746:Vimentin / Q000032:analysis
D000199:Actins; D008301:Maleimides; D014404:Tubulin; D014746:Vimentin; D014316:Tritium; D012701:Serotonin; D000001:Calcimycin; D011493:Protein Kinase C; D009218:Myosins; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium