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Rho GTPases mediated integrin αvβ3 activation in sphingosine-1-phosphate stimulated chemotaxis of endothelial cells

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Abstract

Integrins, a family of transmembrane heterodimeric polypeptides, mediate various biological responses including cell adhesion and migration. In this report, we show that sphingosine-1-phosphate (S1P) activates integrin αvβ3 in endothelial cells (ECs) via the sphingosine-1-phosphate receptor subtype 1 (S1P1)-mediated signaling pathway. S1P treatment results in the activation of integrin αvβ3 in the lamellipodia region of ECs, suggesting that integrin αvβ3 plays a critical role in the S1P-stimulated chemotactic response of ECs. Indeed, S1P treatment induces the association of focal adhesion kinase (FAK) and cytoskeletal proteins with integrin αvβ3, the ligation of αv and β3 subunits, as well as enhances endothelial migration on vitronectin-coated substrata. Knockdown endothelial S1P1 receptor, treatments with pertussis toxin or dominant-negative-Rho family GTPases abrogates the S1P-induced integrin αvβ3 activation in ECs. Consequently, these treatments markedly inhibit the S1P-induced endothelial migratory response on vitronectin-coated substrata. Collectively, these data indicate that the S1P-mediated signaling via the S1P1/Gi/Rho GTPases pathway activates integrin αvβ3, which is indispensable for S1P-stimulated chemotactic response of ECs.

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Abbreviations

S1P:

Sphingosine-1-phosphate

S1P1:

Sphingosine-1-phosphate receptor subtype 1 (old nomenclature EDG-1, endothelial differentiating gene-1)

FAK:

Focal adhesion kinase

GPCR:

G-Protein coupled receptor

HUVECs:

Human umbilical vein endothelial cells

ECs:

Cultured endothelial cells

MOI:

Multiplicity of infection

PTx:

Pertussis toxin

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Acknowledgments

We thank Drs. Robert Gray and Binks Wattenberg for critical discussion. This work is supported by NIH grant R01HL071071 and AHA grant-in-aid 0755245B (M.-J.L).

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Authors and Affiliations

  1. Gheens Center on Aging, Department of Microbiology and Immunology, University of Louisville Health Sciences Center, Louisville, KY, 40202, USA

    Lichun Wang, Jen-Fu Lee & Menq-Jer Lee

  2. Greenebaum Cancer Center, Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, MD, 21201, USA

    Chen-Yong Lin

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  1. Lichun Wang
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  2. Jen-Fu Lee
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Correspondence to Menq-Jer Lee.

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Wang, L., Lee, JF., Lin, CY. et al. Rho GTPases mediated integrin αvβ3 activation in sphingosine-1-phosphate stimulated chemotaxis of endothelial cells. Histochem Cell Biol 129, 579–588 (2008). https://doi.org/10.1007/s00418-008-0389-8

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