PUBLICATION

A β-catenin chromobody-based probe highlights endothelial maturation during vascular morphogenesis in vivo

Authors
Gauvrit, S., Zhao, S., Rothbauer, U., Stainier, D.Y.R.
ID
ZDB-PUB-240608-1
Date
2024
Source
Development (Cambridge, England)   151(11): (Journal)
Registered Authors
Stainier, Didier
Keywords
Chromobody, Endothelium, Live imaging, Nanobody, Vascular morphogenesis, Zebrafish, β-catenin
MeSH Terms
  • Adherens Junctions/metabolism
  • Animals
  • Antigens, CD
  • Cadherins*/genetics
  • Cadherins*/metabolism
  • Endothelial Cells/cytology
  • Endothelial Cells/metabolism
  • Endothelium, Vascular/cytology
  • Endothelium, Vascular/metabolism
  • Morphogenesis*
  • Zebrafish*/embryology
  • Zebrafish*/metabolism
  • Zebrafish Proteins*/genetics
  • Zebrafish Proteins*/metabolism
  • beta Catenin*/metabolism
PubMed
38847494 Full text @ Development
Abstract
Visualization of protein dynamics is a crucial step in understanding cellular processes. Chromobodies, fluorescently labelled single-domain antibodies, have emerged as versatile probes for live cell imaging of endogenous proteins. However, how these chromobodies behave in vivo and how accurately they monitor tissue changes remain poorly explored. Here, we generated an endothelial-specific β-catenin chromobody-derived probe and analyzed its expression pattern during cardiovascular development in zebrafish. Using high-resolution confocal imaging, we show that the chromobody signal correlates with the localization of β-catenin in the nucleus and at cell-cell junctions, and thereby can be used to assess endothelial maturation. Loss of Cadherin 5 strongly affects the localization of the chromobody at the cell membrane, confirming the cadherin-based adherens junction role of β-catenin. Furthermore, using a genetic model to block blood flow, we observed that cell junctions are compromised in most endothelial cells but not in the endocardium, highlighting the heterogeneous response of the endothelium to the lack of blood flow. Overall, our data further expand the use of chromobodies for in vivo applications and illustrate their potential to monitor tissue morphogenesis at high resolution.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping