The Role of Actin Microfilaments in the Down-Regulation of the Degranulation Response in RBL-2H3 Mast Cells

• Published in: The Journal of immunology (1950) Vol. 162; no. 4; pp. 2243 - 2250
• Main Authors: Frigeri, Luciano, Apgar, John R
• Format: Journal Article
• Language: English
• Published: United States Am Assoc Immnol 15-02-1999
• Subjects: Actin Cytoskeleton - drug effects; Actin Cytoskeleton - physiology; Actins - antagonists & inhibitors; Actins - physiology; Animals; Bridged Bicyclo Compounds, Heterocyclic - pharmacology; Cell Degranulation - drug effects; Cell Degranulation - immunology; Cytochalasin D - pharmacology; Dinitrophenols - immunology; Dose-Response Relationship, Immunologic; Down-Regulation - drug effects; Down-Regulation - immunology; Haptens - immunology; Leukemia, Basophilic, Acute; Marine Toxins - pharmacology; Mast Cells - drug effects; Mast Cells - immunology; Mast Cells - physiology; Porifera; Rats; Receptors, IgE - physiology; Serum Albumin, Bovine - immunology; Signal Transduction - drug effects; Signal Transduction - immunology; Thiazoles - pharmacology; Thiazolidines; Tumor Cells, Cultured
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.162.4.2243

Cross-linking of FcepsilonRI on rat basophilic leukemia (RBL) cells initiates a signaling cascade leading to degranulation of the cells and the release of inflammatory mediators. Inhibitors that disrupt microfilaments, such as latrunculin and cytochalasin D, do not cause any degranulation on their own, but they do enhance FcepsilonRI-mediated degranulation. Dose-response studies show a good correlation between inhibition of actin polymerization and increased degranulation. In RBL cells, latrunculin causes a decrease in basal levels of filamentous actin (F-actin), while cytochalasin D does not. This is particularly evident in the Triton-insoluble pool of F-actin which is highly cross-linked and associated with the plasma membrane. A concentration of 500 nM latrunculin decreases the basal level of Triton-insoluble F-actin by 60-70% and total F-actin levels by 25%. Latrunculin increases both the rate and extent of Ag-induced degranulation while having no effect on pervanadate-induced degranulation. Pervanadate activates the signaling pathways directly and bypasses the cross-linking of the receptor. RBL cells, activated through FcepsilonRI in the presence of latrunculin, show increased phospholipase activity as well as increased tyrosine phosphorylation of Syk and increased tyrosine phosphorylation of the receptor itself by the tyrosine kinase Lyn. This indicates that the very earliest signaling events after receptor cross-linking are enhanced. These results suggest that actin microfilaments may interact, either directly or indirectly, with the receptor itself and that they may regulate the signaling process at the level of receptor phosphorylation. Microfilaments may possibly act by uncoupling Lyn from the cross-linked receptor.