A critical role for phospholipase Cgamma2 in alphaIIbbeta3-mediated platelet spreading.
The interaction of fibrinogen with the integrin alphaIIbbeta3 plays a crucial role in platelet adhesion and platelet activation leading to the generation of intracellular signals that nucleate the reorganization of the cytoskeleton. Presently, we have only a limited understanding of the signaling cascades and effector proteins through which changes in the cytoskeletal architecture are mediated. The present study identifies phospholipase Cgamma2 (PLCgamma2) as an important target of the Src-dependent signaling cascade regulated by alphaIIbbeta3. Real time phasecontrast microscopy is used to show that formation of filopodia and lamellapodia in murine platelets on a fibrinogen surface is dramatically inhibited in the absence of PLCgamma2. Significantly, the formation of these structures is mediated by Ca2+ elevation and activation of protein kinase C, both directly regulated by PLC activity. With the involvement of Syk, SLP-76, and Btk, alphaIIbbeta3-induced PLCgamma2 activation partly overlaps with the pathway used by the collagen receptor glycoprotein VI. Important differences, however, exist between the two signaling cascades in that activation of PLCgamma2 by alphaIIbbeta3 is unaltered in murine platelets, which lack the FcR gamma-chain or the adaptor LAT, but is abolished in the presence of cytochalasin D. Therefore, PLCgamma2 plays not only a crucial role in activation of alphaIIbbeta3 by collagen receptors but also in alphaIIbbeta3-mediated responses.
Wonerow
Peter
P
Department of Pharmacology, University of Oxford, Mansfield Road, Oxford OX1 3QT, United Kingdom. peter.wonerow@aventis.com
Pearce
Andrew C
AC
Vaux
David J
DJ
Watson
Steve P
SP
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D001792:Blood Platelets / Q000502:physiology*; D002118:Calcium / Q000378:metabolism; D002352:Carrier Proteins / Q000502:physiology; D015638:Cytochalasin D / Q000494:pharmacology; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D008565:Membrane Proteins; D051379:Mice; D008810:Mice, Inbred C57BL; D051966:Phospholipase C gamma; D010750:Phosphoproteins / Q000502:physiology; D010766:Phosphorylation; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000502:physiology*; D010980:Platelet Membrane Glycoproteins / Q000494:pharmacology; D011505:Protein-Tyrosine Kinases / Q000502:physiology; D010738:Type C Phospholipases / Q000502:physiology*
D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; C491734:Lat protein, mouse; D008565:Membrane Proteins; D010750:Phosphoproteins; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D010980:Platelet Membrane Glycoproteins; C098341:platelet membrane glycoprotein VI; D015638:Cytochalasin D; D011505:Protein-Tyrosine Kinases; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; C496051:Ptk2 protein, mouse; D010738:Type C Phospholipases; D051966:Phospholipase C gamma; D002118:Calcium