Microvessel formation from mouse embryonic aortic explants is oxygen and VEGF dependent.
To delineate the roles of O(2) and vascular endothelial growth factor (VEGF) in the process of angiogenesis from the embryonic aorta, we cultured mouse embryonic aorta explants (thoracic level to lateral vessels supplying the mesonephros and metanephros) in a three-dimensional type I collagen gel matrix. During 8 days of culture under 5% O(2), but not room air, the addition of VEGF to explants stimulated the formation of CD31-positive, Flk-1-positive, Gs-IB(4)-positive structures in a concentration-dependent manner. Electron microscopy showed the structures to be capillary-like. VEGF-induced capillary-like structure formation was inhibited by sequestration of VEGF via addition of soluble Flt-1 fusion protein or anti-VEGF antibodies. Expression of Flk-1, but not Flt-1, was increased in embryonic aorta cultured under 5% O(2) relative to room air. Our data suggest that low O(2) upregulates Flk-1 expression in embryonic aorta in vitro and renders it more responsive to VEGF.
George M. O'Brien Kidney and Urological Disease Center, Renal Division, Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Tetsu
T
Akimoto
Helen
H
Liapis
Marc R
MR
Hammerman
D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D000943:Antigens, Differentiation / Q000096:biosynthesis; D001011:Aorta / Q000166:cytology / Q000187:drug effects* / Q000196:embryology; D002196:Capillaries / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D015687:Cell Hypoxia / Q000502:physiology; D046508:Culture Techniques; D004305:Dose-Response Relationship, Drug; D016228:Endothelial Growth Factors / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D008222:Lymphokines / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D051379:Mice; D008810:Mice, Inbred C57BL; D018919:Neovascularization, Physiologic / Q000187:drug effects* / Q000502:physiology; D010100:Oxygen / Q000494:pharmacology / Q000502:physiology*; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000096:biosynthesis; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis; D020794:Receptor Protein-Tyrosine Kinases / Q000096:biosynthesis / Q000378:metabolism; D017978:Receptors, Growth Factor / Q000378:metabolism; D040262:Receptors, Vascular Endothelial Growth Factor; D011993:Recombinant Fusion Proteins / Q000494:pharmacology; D042461:Vascular Endothelial Growth Factor A; D040281:Vascular Endothelial Growth Factor Receptor-1; D042442:Vascular Endothelial Growth Factors
D000906:Antibodies; D000943:Antigens, Differentiation; D016228:Endothelial Growth Factors; D008222:Lymphokines; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D011518:Proto-Oncogene Proteins; D017978:Receptors, Growth Factor; D011993:Recombinant Fusion Proteins; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D020794:Receptor Protein-Tyrosine Kinases; D040262:Receptors, Vascular Endothelial Growth Factor; D040281:Vascular Endothelial Growth Factor Receptor-1; D010100:Oxygen