Increased Expression of Axl Tyrosine Kinase After Vascular Injury and Regulation by G Protein–Coupled Receptor Agonists in Rats

From the Department of Medicine, Division of Cardiovascular Research, University of Washington (M.G.M., D.A.W., V.P., D.G., B.C.B., M.A.C.), Seattle; and Maine Medical Center Research Institute (V.L.), South Portland.

Correspondence to Marshall A. Corson, MD, Box 357710, Seattle, WA 98195-7710. E-mail mcorson{at}u.washington.edu

Abstract—Axl is a receptor tyrosine kinase originally identified as a transforming gene product in human myeloid leukemia cells. Cultured rat vascular smooth muscle cells also express Axl, where it has been proposed that Axl may play a role in cell proliferation. In the current study, we tested the hypotheses that Axl expression would parallel neointima formation in balloon-injured rat carotid, and that Axl expression would be regulated by growth factors present at sites of vascular injury. Ribonuclease protection assay showed dynamic increases in Axl mRNA in vessels, with peak expression 7 and 14 days after injury. Immunohistochemical analysis confirmed these results and demonstrated that Axl protein expression was localized primarily to cells of the neointima after injury. Northern blot analysis indicated increased mRNA expression for the secreted Axl ligand, Gas6, in injured carotids, with a time course paralleling that of Axl upregulation. Axl and Gas6 expression were temporally correlated with neointima formation, suggesting a role for Axl signaling in this process. Other studies, performed in cultured rat vascular smooth muscle cells, revealed positive regulation of Axl mRNA expression by thrombin or angiotensin II but not by basic fibroblast growth factor, platelet-derived growth factor-BB, or transforming growth factor-ß₁. Western blot analysis confirmed these results, showing that Axl protein expression was specifically increased by thrombin or angiotensin II. Our results implicate Axl as a potential mediator of vascular smooth muscle migration and proliferation caused by vascular injury and G protein–coupled receptor agonists.

Key Words: angiotensin II • thrombin • proliferation • neointima • vascular smooth muscle

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