Effects of VEGF(165) and VEGF(121) on vasculogenesis and angiogenesis in cultured embryonic quail hearts.
It has been documented that hypoxia enhances coronary vasculogenesis and angiogenesis in cultured embryonic quail hearts via the upregulation of vascular endothelial growth factor (VEGF). In this study, we compared the functions of two VEGF splice variants. Ventricles from 6-day-old embryonic quail hearts were cultured on three-dimensional collagen gels. Recombinant human VEGF(121) or VEGF(165) were added to the culture medium for 48 h, and vascular growth was visualized by immunostaining with a quail-specific endothelial cell (EC) marker, QH1. VEGF(165) enhanced vascular growth in a dose-dependent manner: 5 ng/ml of VEGF(165) slightly increased the number of ECs, 10 ng/ml of VEGF(165) increased the incorporation of ECs into tubular structures, and at 20 ng/ml of VEGF(165) wider tubes were formed. This pattern plateaued at the 50 ng/ml dose. In contrast, VEGF(121) did not enhance either the number of ECs or tube formation at these or higher dosages. Combined effects of hypoxia and exogenous VEGF(165) were then compared. Tube formation from the heart explants treated with both hypoxia and 50 ng/ml of VEGF(165) had a morphology intermediate to those treated with hypoxia or VEGF(165) alone. Immunocytochemistry study revealed EC lumenization under all culture conditions. However, the addition of VEGF(165) stimulated the coalescence of ECs to form larger vessels. We conclude the following: 1) VEGF(121) and VEGF(165) induced by hypoxia have different functions on coronary vascular growth, 2) unknown factors induced by hypoxia can modify the effect of VEGF(165), and 3) EC lumenization observed in the heart explant culture closely mimics in vivo coronary vasculogenesis.
Yue
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Department of Anatomy and Cell Biology and the Cardiovascular Center, University of Iowa, Iowa City, Iowa 52242, USA.
Tomanek
R J
RJ
D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002642:Chick Embryo; D003326:Coronary Circulation / Q000187:drug effects*; D016228:Endothelial Growth Factors / Q000494:pharmacology*; D004730:Endothelium, Vascular / Q000648:ultrastructure; D006321:Heart / Q000196:embryology; D000860:Hypoxia / Q000503:physiopathology; D008222:Lymphokines / Q000494:pharmacology*; D008854:Microscopy, Electron; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D009924:Organ Culture Techniques; D010100:Oxygen / Q000494:pharmacology; D011784:Quail; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D016228:Endothelial Growth Factors; D008222:Lymphokines; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D010100:Oxygen