title: Dynamics of VEGF matrix-retention in vascular network patterning.
abstract: Vascular endothelial growth factor (VEGF) is a central regulator of blood vessel morphogenesis, although its role in patterning of endothelial cells into vascular networks is not fully understood. It has been suggested that binding of soluble VEGF to extracellular matrix components causes spatially restricted cues that guide endothelial cells into network patterns. Yet, current evidence for such a mechanism remains indirect. In this study, we quantitatively analyse the dynamics of VEGF retention in a controlled in vitro situation of human umbilical vascular endothelial cells (HUVECs) in Matrigel. We show that fluorescent VEGF accumulates in pericellular areas and colocalizes with VEGF binding molecules. Analysis of fluorescence recovery after photobleaching reveals that binding/unbinding to matrix molecules dominates VEGF dynamics in the pericellular region. Computational simulations using our experimental measurements of kinetic parameters show that matrix retention of chemotactic signals can lead to the formation of reticular cellular networks on a realistic timescale. Taken together, these results show that VEGF binds to matrix molecules in proximity of HUVECs in Matrigel, and suggest that bound VEGF drives vascular network patterning.
authors.affiliation: Faculty of Mathematics, Department of Applied Mathematics, Universidad Complutense de Madrid, E-28040 Madrid, Spain. Department for Innovative Methods of Computing, Center for Information Services and High Performance Computing (ZIH), Technische Universität Dresden, D-01062 Dresden, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Köhn-Luque
authors.forename: W
authors.initials: W
authors.lastname: de Back
authors.forename: Y
authors.initials: Y
authors.lastname: Yamaguchi
authors.forename: K
authors.initials: K
authors.lastname: Yoshimura
authors.forename: M A
authors.initials: MA
authors.lastname: Herrero
authors.forename: T
authors.initials: T
authors.lastname: Miura
mesh_terms: D001672:Biocompatible Materials / Q000737:chemistry; D003094:Collagen / Q000737:chemistry; D003198:Computer Simulation; D004338:Drug Combinations; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology* / Q000378:metabolism*; D006801:Humans; D007797:Laminin / Q000737:chemistry; D008954:Models, Biological; D018919:Neovascularization, Physiologic; D011485:Protein Binding; D011509:Proteoglycans / Q000737:chemistry; D042461:Vascular Endothelial Growth Factor A / Q000032:analysis / Q000378:metabolism*
chemical_list: D001672:Biocompatible Materials; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; D042461:Vascular Endothelial Growth Factor A; C059485:matrigel; D003094:Collagen