α-Adrenergic-mediated activation of human reconstituted fibrinogen receptor (integrin αIIbβ3) in Chinese hamster ovary cells

Nora Butta; Susana Larrucea; Consuelo Gonzalez-Manchon; Sonia Alonso; Roberto Parrilla

doi:10.1160/TH04-02-0090

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2004; 92(06): 1368-1376
DOI: 10.1160/TH04-02-0090

Platelets and Blood Cells

Schattauer GmbH

α-Adrenergic-mediated activation of human reconstituted fibrinogen receptor (integrin αIIbβ3) in Chinese hamster ovary cells

Nora Butta

¹Department of Physiopathology and Human Molecular Genetics

²Centro de Investigaciones Biológicas (CSIC), Madrid, Spain

,

Susana Larrucea

¹Department of Physiopathology and Human Molecular Genetics

²Centro de Investigaciones Biológicas (CSIC), Madrid, Spain

,

Consuelo Gonzalez-Manchon

¹Department of Physiopathology and Human Molecular Genetics

²Centro de Investigaciones Biológicas (CSIC), Madrid, Spain

,

Sonia Alonso

¹Department of Physiopathology and Human Molecular Genetics

²Centro de Investigaciones Biológicas (CSIC), Madrid, Spain

,

Roberto Parrilla

¹Department of Physiopathology and Human Molecular Genetics

²Centro de Investigaciones Biológicas (CSIC), Madrid, Spain

› Institutsangaben

Abstract

Summary

This work reports the functional studies of CHO cells coexpressing α-adrenergic (αAR) and human fibrinogen (Fg) receptors (integrin αIIbβ3). Stimulation of these cells with α-agonists produced a transient rise in the free cytosolic calcium (Ca⁺⁺) accompanied by enhanced binding to soluble Fg, and these effects were prevented by specific αAR antagonists. The αadrenergic-induced activation of αIIbβ3 in CHO-αIIbβ3-αAR increased the rate of adhesion and extension of cells onto Fg coated plates, and also induced a soluble Fgand αIIbβ3-dependent formation of cell aggregates, whereas no effects were observed by the stimulation of CHO-αIIbβ3 cells. α-Adrenergic antagonists, the ligand mimetic peptide RGDS, pertussis toxin (PTX), or EDTA, they all prevented the α-adrenergic stimulation of adhesion and aggregation. However, inhibition of PKC prevented the α-adrenergic stimulation of cell adherence, whereas blocking the intracellular Ca⁺⁺ mobilization impeded the stimulation of cell aggregation. The α-adrenergic activation was associated with phosphorylation of a protein of ∼100 kDa and proteins of the MAPK family. The former was selectively phosphorylated by α-adrenergic stimulation whereas the latter were phosphorylated by the binding of cells to Fg and markedly intensified by α-adrenergic stimulation.

Keywords

α-adrenergic receptors - adhesion receptors/integrins - fibrinogen/fibrin - gene expression - molecular biology methods

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Referenzen

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