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Single Cell Analysis of Endothelial Morphogenesis during Sprouting Angiogenesis

Friday, November 08, 2013 — Poster Session IV

2:00 p.m. – 4:00 p.m.

FAES Academic Center (Upper-Level Terrace)




  • JA Yu
  • BM Weinstein


Angiogenesis is critical for vertebrate organogenesis and plays key roles in pathological processes such as cancer. Although vessel formation has been extensively studied at the tissue level, lack of proper genetic tools and difficulty in in vivo imaging have hampered study of dynamic cell behaviors during endothelial morphogenesis. We have now developed endothelium-specific fluorescent reporter transgenic zebrafish and high-speed two-photon imaging methods allowing us to examine in vivo EC morphogenesis at single cell resolution. New transgenic tools have been generated to simultaneously mark EC nuclei, membranes, and junctions with different fluorescent proteins. These transgenes, which we are using in injected mosaics, permit definitive identification and imaging of individual ECs during vessel sprouting and lumenization. Combined with sophisticated two-photon imaging methods that provide an unprecedented new level of resolution, we are using these tools and methods to probe the morphological rearrangements of individual ECs during trunk intersegmental vessel sprouting and examine intracellular and extracellular modes of lumen formation. The genetic tools and methods developed here, and the approach of single cell analysis, will be useful not only for defining the concerted EC behaviors during normal vessel formation, but for examining the abnormal endothelial phenotype in zebrafish models of human vascular disease.

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