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Using En Face Immunofluorescence Staining to Observe Vascular Endothelial Cells Directly
06:09

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Published on: August 20, 2019

AFM functional imaging on vascular endothelial cells.

Lilia A Chtcheglova1, Linda Wildling, Jens Waschke

  • 1Institute for Biophysics, Johannes Kepler University of Linz, A-4040 Linz, Austria. lilia.chtcheglova@jku.at

Journal of Molecular Recognition : JMR
|November 2, 2010
PubMed
Summary
This summary is machine-generated.

Vascular endothelial (VE)-cadherin forms cell connections, visualized using nano-mapping. This technique reveals VE-cadherin clusters linked to the actin cytoskeleton in endothelial cells.

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Area of Science:

  • Cell Biology
  • Biophysics
  • Nanotechnology

Background:

  • Vascular endothelial (VE)-cadherin mediates mechanical linkage between endothelial cells.
  • VE-cadherin forms clusters linked to the actin cytoskeleton, a dynamic process influenced by cell function.

Purpose of the Study:

  • To nano-map VE-cadherin distribution and clustering in mouse myocardial microvascular endothelial cells (MyEnd).
  • To investigate the relationship between VE-cadherin clusters and the actin cytoskeleton.

Main Methods:

  • Simultaneous topography and recognition imaging (TREC) technique was employed.
  • TREC provided high lateral resolution (nanometer scale) mapping of VE-cadherin on cell surfaces.
  • Microtubule depolymerization using nocodazole was performed to assess cytoskeletal interactions.

Main Results:

  • TREC revealed prominent VE-cadherin 'dark' spots (10-250 nm domains/clusters) arising from cis-dimer binding.
  • These domains collocalized with cytoskeletal filaments, even after microtubule depolymerization.
  • VE-cadherin clusters were confirmed to be linked to actin filaments.

Conclusions:

  • TREC is an effective method for high-resolution mapping of cell surface receptors.
  • VE-cadherin is dynamically linked to the actin cytoskeleton in endothelial cells.
  • The findings support the hypothesis of VE-cadherin's direct association with actin filaments.