Phosphatidylinositol 3,4,5-trisphosphate-dependent stimulation of phospholipase C-gamma2 is an early key event in FcgammaRIIA-mediated activation of human platelets.
Platelets express a single class of Fcgamma receptor (FcgammaRIIA), which is involved in heparin-associated thrombocytopenia and possibly in inflammation. FcgammaRIIA cross-linking induces platelet secretion and aggregation, together with a number of cellular events such as tyrosine phosphorylation, activation of phospholipase C-gamma2 (PLC-gamma2), and calcium signaling. Here, we show that in response to FcgammaRIIA cross-linking, phosphatidylinositol (3,4, 5)-trisphosphate (PtdIns(3,4,5)P3) is rapidly produced, whereas phosphatidylinositol (3,4)-bisphosphate accumulates more slowly, demonstrating a marked activation of phosphoinositide 3-kinase (PI 3-kinase). Inhibition of PI 3-kinase by wortmannin or LY294002 abolished platelet secretion and aggregation, as well as phospholipase C (PLC) activation, indicating a role of this lipid kinase in the early phase of platelet activation. Inhibition of PLCgamma2 was not related to its tyrosine phosphorylation state, since wortmannin actually suppressed its dephosphorylation, which requires platelet aggregation and integrin alphaIIb/beta3 engagement. In contrast, the stable association of PLCgamma2 to the membrane/cytoskeleton interface observed at early stage of platelet activation was fully abolished upon inhibition of PI 3-kinase. In addition, PLCgamma2 was able to preferentially interact in vitro with PtdIns(3,4,5)P3. Finally, exogenous PtdIns(3,4,5)P3 restored PLC activation in permeabilized platelets treated with wortmannin. We propose that PI 3-kinase and its product PtdIns(3,4,5)P3 play a key role in the activation and adequate location of PLCgamma2 induced by FcgammaRIIA cross-linking.
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Institut Fédératif de Recherche en Immunologie Cellulaire et Moléculaire, Université Paul Sabatier and Centre Hospitalo-Universitaire de Toulouse, INSERM, Unité 326, France.
Payrastre
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Viala
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Mauco
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Plantavid
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D000730:Androstadienes / Q000494:pharmacology; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D015703:Antigens, CD / Q000187:drug effects / Q000502:physiology*; D001792:Blood Platelets / Q000187:drug effects / Q000276:immunology / Q000502:physiology*; D002462:Cell Membrane / Q000502:physiology; D002867:Chromones / Q000494:pharmacology; D003599:Cytoskeleton / Q000502:physiology; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000097:blood*; D007700:Kinetics; D008954:Models, Biological; D009025:Morpholines / Q000494:pharmacology; D018129:Phosphatidylinositol Phosphates / Q000096:biosynthesis / Q000097:blood*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D051966:Phospholipase C gamma; D015539:Platelet Activation / Q000187:drug effects / Q000502:physiology*; D017452:Receptors, IgG / Q000187:drug effects / Q000502:physiology*; D012701:Serotonin / Q000097:blood; D015398:Signal Transduction; D013917:Thrombin / Q000494:pharmacology; D010738:Type C Phospholipases / Q000097:blood*; D000077191:Wortmannin
D000730:Androstadienes; D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D002867:Chromones; D004791:Enzyme Inhibitors; C080568:Fc gamma receptor IIA; D007527:Isoenzymes; D009025:Morpholines; D018129:Phosphatidylinositol Phosphates; D000081082:Phosphoinositide-3 Kinase Inhibitors; D017452:Receptors, IgG; C060974:phosphatidylinositol 3,4,5-triphosphate; C060980:phosphatidylinositol 3,4-diphosphate; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D012701:Serotonin; D010738:Type C Phospholipases; D051966:Phospholipase C gamma; D013917:Thrombin; D000077191:Wortmannin