Role of phosphatidylinositol 3-kinase in degranulation induced by IgE-dependent and -independent mechanisms in rat basophilic RBL-2H3 (ml) cells.
We have examined the role of phosphatidylinositol 3-kinase (P13-kinase) in the degranulation induced by the antigen, an IgE-dependent stimulant, and by carbachol and thapsigargin, IgE-independent stimulants, in the muscarine ml receptor-transfected mast cell line RBL-2h3 (ml) cells. These stimulants commonly increased P13-kinase activity in the anti-phosphotyrosine immunoprecipitate. The P13-kinase inhibitors wortmannin and LY294002 inhibited induced by these stimulants. The membrane ruffling induced by the antigen or carbachol was also inhibited by wortmannin. In contrast, thapsigargin induced by membrane ruffling but induced microspikes, which was not affected by wortmannin. In the permeabilized RBL-2H3 (ml) cells, wortmannin the GTP gamma S-induced membrane ruffling without inhibiting the GTP gamma S-induced degranulation. These findings suggest that P13-kinase is involved not only in IgE-dependent degranulation but also in IgE-independent degranulation, and that the GTP gamma S-sensitive protein at the downstream of P13-kinase is responsible for the degranulation but not for the membrane ruffling.
Hirasawa
N
N
Department of Pathophysiological Biochemistry, Faculty of Pharmaceutical Sciences, Tohoku University, Miyagi, Japan.
Sato
Y
Y
Yomogida
S
S
Mue
S
S
Ohuchi
K
K
D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D000941:Antigens / Q000494:pharmacology; D002217:Carbachol / Q000494:pharmacology; D015550:Cell Degranulation / Q000502:physiology*; D002461:Cell Line, Transformed; D002463:Cell Membrane Permeability; D002867:Chromones / Q000494:pharmacology; D004140:Dinitrophenols / Q000494:pharmacology; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D007073:Immunoglobulin E / Q000378:metabolism*; D008407:Mast Cells / Q000187:drug effects / Q000201:enzymology* / Q000502:physiology; D009025:Morpholines / Q000494:pharmacology; D019869:Phosphatidylinositol 3-Kinases; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000502:physiology*; D051381:Rats; D012709:Serum Albumin / Q000494:pharmacology; D019284:Thapsigargin / Q000494:pharmacology; D000077191:Wortmannin; D001619:beta-N-Acetylhexosaminidases / Q000378:metabolism
D000730:Androstadienes; D000941:Antigens; D002867:Chromones; D004140:Dinitrophenols; D004791:Enzyme Inhibitors; D009025:Morpholines; D012709:Serum Albumin; C058064:dinitrophenyl-human serum albumin conjugate; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D007073:Immunoglobulin E; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D019284:Thapsigargin; D002217:Carbachol; D019869:Phosphatidylinositol 3-Kinases; D017853:Phosphotransferases (Alcohol Group Acceptor); D001619:beta-N-Acetylhexosaminidases; D000077191:Wortmannin