Impaired "outside-in" integrin alphaIIbbeta3 signaling and thrombus stability in TSSC6-deficient mice.

We investigated the role of the hematopoietic-specific tetraspanin superfamily member, TSSC6, in platelet function using wild-type mice and TSSC6-deficient mice. TSSC6 is expressed on the surface of murine platelets and is up-regulated by thrombin stimulation, indicating an intracellular pool of TSSC6. Immunoprecipitation/Western blot studies reveal a constitutive physical association of TSSC6 with the integrin alpha(IIb)beta(3) complex under strong detergent conditions. In vivo evaluation of hemostasis by tail bleeding revealed increased bleeding time, volume of blood lost, and evidence of tail rebleeds in TSSC6 null mice, indicating unstable hemostasis. Using ex vivo techniques, we showed that TSSC6-deficient platelets exhibited impaired kinetics of clot retraction, platelet aggregation at lower doses of PAR-4, and collagen and platelet spreading on fibrinogen in the presence of normal integrin alpha(IIb)beta(3) expression. TSSC6-deficient platelets showed normal alpha granule secretion, normal "inside-out" integrin alpha(IIb)beta(3) signaling (fluorescein isothiocyanate [FITC]-fibrinogen and JON/A binding), and normal platelet adhesion on fibrinogen. Furthermore, we show that absence of platelet TSSC6 affects the secondary stability of arterial thrombi in vivo upon vascular injury. These data demonstrate that TSSC6 appears to regulate integrin alpha(IIb)beta(3) "outside-in" signaling events in platelets and is necessary for stability of arterial thrombi in vivo.

[1]  L. Brass,et al.  Minding the gaps to promote thrombus growth and stability. , 2005, The Journal of clinical investigation.

[2]  N. Prévost,et al.  Eph kinases and ephrins support thrombus growth and stability by regulating integrin outside-in signaling in platelets. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[3]  S. Israels,et al.  CD63 modulates spreading and tyrosine phosphorylation of platelets on immobilized fibrinogen , 2004, Thrombosis and Haemostasis.

[4]  M. Wright,et al.  The tetraspanin superfamily member CD151 regulates outside-in integrin αIIbβ3 signaling and platelet function , 2004 .

[5]  M. Wright,et al.  Characterization of Mice Lacking the Tetraspanin Superfamily Member CD151 , 2004, Molecular and Cellular Biology.

[6]  M. Wright,et al.  A Regulatory Role for CD37 in T Cell Proliferation1 , 2004, The Journal of Immunology.

[7]  R. Alon,et al.  The CD81 Tetraspanin Facilitates Instantaneous Leukocyte VLA-4 Adhesion Strengthening to Vascular Cell Adhesion Molecule 1 (VCAM-1) under Shear Flow* , 2003, Journal of Biological Chemistry.

[8]  D. Jackson,et al.  Physical proximity and functional interplay of PECAM-1 with the Fc receptor Fc gamma RIIa on the platelet plasma membrane. , 2003, Blood.

[9]  Richard T. Lee,et al.  Tetraspanin CD151 regulates α6β1 integrin adhesion strengthening , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[10]  T. V. Kolesnikova,et al.  Functional domains in tetraspanin proteins. , 2003, Trends in biochemical sciences.

[11]  M. Wright,et al.  The Absence of Tssc6, a Member of the Tetraspanin Superfamily, Does Not Affect Lymphoid Development but Enhances In Vitro T-Cell Proliferative Responses , 2002, Molecular and Cellular Biology.

[12]  G. Freeman,et al.  Critical role of CD81 in cognate T-B cell interactions leading to Th2 responses. , 2002, International immunology.

[13]  R. Hynes,et al.  CD40L stabilizes arterial thrombi by a β3 integrin–dependent mechanism , 2002, Nature Medicine.

[14]  M. Hemler,et al.  Specific tetraspanin functions , 2001, The Journal of cell biology.

[15]  F. Berditchevski.  Complexes of tetraspanins with integrins: more than meets the eye. , 2001, Journal of cell science.

[16]  M. Wright,et al.  Molecular characterisation of mouse and human TSSC6: evidence that TSSC6 is a genuine member of the tetraspanin superfamily and is expressed specifically in haematopoietic organs. , 2001, Biochimica et biophysica acta.

[17]  H. Liu,et al.  Anti-human platelet tetraspanin (CD9) monoclonal antibodies induce platelet integrin alphaIIbbeta3 activation in a Fc receptor-independent fashion. , 2000, Chinese medical sciences journal = Chung-kuo i hsueh k'o hsueh tsa chih.

[18]  M. Ko,et al.  Phemx, a novel mouse gene expressed in hematopoietic cells maps to the imprinted cluster on distal chromosome 7. , 2000, Genomics.

[19]  S. Oda,et al.  The gamete fusion process is defective in eggs of Cd9-deficient mice , 2000, Nature Genetics.

[20]  C. Jasmin,et al.  Severely reduced female fertility in CD9-deficient mice. , 2000, Science.

[21]  E. Mekada,et al.  Requirement of CD9 on the egg plasma membrane for fertilization. , 2000, Science.

[22]  N. Killeen,et al.  Integrin cytoplasmic tyrosine motif is required for outside-in αIIbβ3 signalling and platelet function , 1999, Nature.

[23]  S. Goodman,et al.  Sheep, pig, and human platelet-material interactions with model cardiovascular biomaterials. , 1999, Journal of biomedical materials research.

[24]  A. Strasser,et al.  Rapid hybridoma screening method for the identification of monoclonal antibodies to low-abundance cytoplasmic proteins. , 1998, BioTechniques.

[25]  R. Hynes,et al.  A mouse model of severe von Willebrand disease: defects in hemostasis and thrombosis. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[26]  K. Campbell,et al.  Normal development but differentially altered proliferative responses of lymphocytes in mice lacking CD81 , 1997, The EMBO journal.

[27]  Y. Yatomi,et al.  Differential activation of human platelets induced by Fc gamma receptor II cross-linking and by anti-CD9 monoclonal antibody. , 1996, Journal of immunology.

[28]  Y. Yatomi,et al.  Fc gamma II receptor-mediated platelet activation induced by anti-CD9 monoclonal antibody opens Ca2+ channels which are distinct from those associated with Ca2+ store depletion. , 1995, Journal of immunology.

[29]  M. Berndt,et al.  Platelet alpha-granule and plasma membrane share two new components: CD9 and PECAM-1 , 1994 .

[30]  J. Slupsky,et al.  Evidence that monoclonal antibodies against CD9 antigen induce specific association between CD9 and the platelet glycoprotein IIb-IIIa complex. , 1989, The Journal of biological chemistry.

[31]  D. Fitzpatrick,et al.  Multiple pulmonary emboli in a patient with afibrinogenaemia. , 1988, Acta haematologica.

[32]  A. Mulder,et al.  Megakaryoblastic differentiation of proerythroblastic K562 cell-line cells. , 1984, Leukemia research.

[33]  G. Ingram,et al.  Fatal pulmonary embolus in congenital fibrinopenia. Report of two cases. , 1966, Acta haematologica.