© 1995 American Heart Association, Inc.
Articles |
Osteopontin and ß3 Integrin Are Coordinately Expressed in Regenerating Endothelium In Vivo and Stimulate Arg-Gly-Asp–Dependent Endothelial Migration In Vitro
From the Department of Pathology (V.L., S.M.S., C.M.G.), University of Washington, Seattle; the London (Canada) Regional Cancer Centre (A.F.C.); and the Department of Cell Biology (L.L.), Vanderbilt University, Nashville, Tenn.
Abstract Osteopontin is an Arg-Gly-Asp (RGD)–containing
acidic glycoprotein postulated to mediate cellular adhesion
and migration in a growing number of normal and pathological conditions
through interaction with integrin molecules. In this report, we have
investigated the potential contributions of osteopontin and one of its
receptors, the 
vß3 integrin, to
endothelial regenerative processes by using both in
vivo and in vitro models. In vivo, uninjured rat arterial
endothelium had undetectable levels of osteopontin and
ß3-integrin mRNA by in situ hybridization. After balloon
catheter denudation, osteopontin mRNA levels correlated temporally and
spatially with active endothelial proliferation and
migration, with the highest levels observed at the wound edge between 8
hours and 2 weeks after injury, declining to uninjured levels at 6
weeks, when regeneration was complete. Osteopontin protein levels, as
determined by immunocytochemistry, paralleled the time course
of mRNA expression. Likewise, ß3-integrin mRNA and
protein levels were substantially elevated in regenerating
endothelial cells but were not detectable in uninjured
or healed endothelium. In vitro, rat smooth muscle
cell–derived and bacterial expressed mouse recombinant
osteopontins both stimulated the adhesion and directed migration of
bovine aortic endothelial cells through interactions
with the vß3 receptor. Structural mutants
of osteopontin confirmed the importance of the RGD domain for both
adhesion and migration of endothelial cells through
vß3. These data suggest important roles
for osteopontin and ß3 integrin in regenerating
endothelium.
Key Words: osteopontin • integrin • endothelium • adhesion • migration
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