title: Single-cell analysis of endothelial morphogenesis in vivo.
abstract: Vessel formation has been extensively studied at the tissue level, but the difficulty in imaging the endothelium with cellular resolution has hampered study of the morphogenesis and behavior of endothelial cells (ECs) in vivo. We are using endothelial-specific transgenes and high-resolution imaging to examine single ECs in zebrafish. By generating mosaics with transgenes that simultaneously mark endothelial nuclei and membranes we are able to definitively identify and study the morphology and behavior of individual ECs during vessel sprouting and lumen formation. Using these methods, we show that developing trunk vessels are composed of ECs of varying morphology, and that single-cell analysis can be used to quantitate alterations in morphology and dynamics in ECs that are defective in proper guidance and patterning. Finally, we use single-cell analysis of intersegmental vessels undergoing lumen formation to demonstrate the coexistence of seamless transcellular lumens and single or multicellular enclosed lumens with autocellular or intercellular junctions, suggesting that heterogeneous mechanisms contribute to vascular lumen formation in vivo. The tools that we have developed for single EC analysis should facilitate further rigorous qualitative and quantitative analysis of EC morphology and behavior in vivo.
authors.affiliation: Program in Genomics of Differentiation, The Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Jianxin A
authors.initials: JA
authors.lastname: Yu
authors.affiliation: Program in Genomics of Differentiation, The Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Castranova
authors.affiliation: Program in Genomics of Differentiation, The Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Van N
authors.initials: VN
authors.lastname: Pham
authors.affiliation: Program in Genomics of Differentiation, The Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA weinsteb@mail.nih.gov.
authors.forename: Brant M
authors.initials: BM
authors.lastname: Weinstein
mesh_terms: D000818:Animals; D016764:Cell Polarity; D004625:Embryo, Nonmammalian / Q000166:cytology; D042783:Endothelial Cells / Q000166:cytology; D004727:Endothelium / Q000098:blood supply / Q000166:cytology* / Q000196:embryology*; D021621:Imaging, Three-Dimensional; D007365:Intercellular Junctions; D042541:Intracellular Space / Q000378:metabolism; D008561:Membrane Fusion; D009024:Morphogenesis; D018919:Neovascularization, Physiologic; D015203:Reproducibility of Results; D059010:Single-Cell Analysis / Q000379:methods*; D060726:Torso / Q000098:blood supply / Q000196:embryology; D014617:Vacuoles / Q000378:metabolism; D015027:Zebrafish / Q000196:embryology*
keywords: Claudin 5; Endothelial morphogenesis; Lumenization; Morphodynamics; Plexin D1; Single-cell analysis; Tight junction