ZFIN ID: ZDB-PUB-150809-9
Single cell analysis of endothelial morphogenesis in vivo
Yu, J.A., Castranova, D., Pham, V.N., Weinstein, B.M.
Date: 2015
Source: Development (Cambridge, England)   142(17): 2951-61 (Journal)
Registered Authors: Castranova, Dan, Pham, Van, Weinstein, Brant M., Yu, Jianxin Alex
Keywords: Endothelial morphogenesis, Single-cell analysis, Plexin D1, Morphodynamics, Lumenization, Tight junction, Claudin 5
MeSH Terms:
  • Animals
  • Cell Polarity
  • Embryo, Nonmammalian/cytology
  • Endothelial Cells/cytology
  • Endothelium/blood supply
  • Endothelium/cytology*
  • Endothelium/embryology*
  • Imaging, Three-Dimensional
  • Intercellular Junctions
  • Intracellular Space/metabolism
  • Membrane Fusion
  • Morphogenesis*
  • Neovascularization, Physiologic
  • Reproducibility of Results
  • Single-Cell Analysis/methods*
  • Torso/blood supply
  • Torso/embryology
  • Vacuoles/metabolism
  • Zebrafish/embryology*
PubMed: 26253401 Full text @ Development
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ABSTRACT
Vessel formation has been extensively studied at the tissue level, but difficulty in imaging the endothelium with cellular resolution has hampered study of the morphogenesis and behavior of endothelial cells (EC) in vivo. We are using endothelial-specific transgenes and high-resolution imaging to examine single endothelial cells in the 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 EC during vessel sprouting and lumen formation. Using these methods, we show that developing trunk vessels are composed of EC of varying morphology, and that single cell analysis of EC can be used to quantitate alterations in morphology and dynamics in EC defective in proper guidance and patterning. Finally, we use single cell analysis of intersegmental vessels (ISV) undergoing lumen formation to demonstrate the coexistence of seamless transcellular lumens and single or multicellular enclosed lumens with autocellular or intercellular junctions, suggesting heterogeneous mechanisms contribute to vascular lumen formation in vivo. The tools we have developed for single EC analysis should facilitate further rigorous qualitative and quantitative analysis of EC morphology and behavior in vivo.
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