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Simultaneously Capturing Real-time Images in Two Emission Channels Using a Dual Camera Emission Splitting System: Applications to Cell Adhesion
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Focal Adhesions Undergo Longitudinal Splitting into Fixed-Width Units.

Lorna E Young1, Henry N Higgs1

  • 1Department of Biochemistry and Cell Biology, Geisel School of Medicine at Dartmouth, Hanover, NH 03755, USA.

Current Biology : CB
|June 19, 2018
PubMed
Summary

Focal adhesions (FAs) assemble via splitting of linear units, revealing a fundamental fixed-width unit. This process is influenced by cell type, matrix, and proteins like APC and VASP, impacting cell motility.

Keywords:
collagenfocal adhesion plaquefocal adhesion unitmyosinnano-architecture

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Area of Science:

  • Cell Biology
  • Biophysics
  • Cytoskeleton Dynamics

Background:

  • Focal adhesions (FAs) and stress fibers (SFs) are crucial for cell motility and environmental sensing.
  • The assembly and maturation of FAs and SFs during initial cell spreading remain poorly understood.

Purpose of the Study:

  • To investigate the de novo assembly and maturation process of focal adhesions.
  • To identify the fundamental structural units and dynamic behaviors of focal adhesions during cell spreading.

Main Methods:

  • Live-cell Airyscan microscopy was employed to visualize FA and SF dynamics.
  • Experimental manipulation involved depletion of adenomatous polyposis coli (APC) and vasodilator-stimulated phosphoprotein (VASP).

Main Results:

  • Focal adhesions were observed to undergo a "splitting" process during assembly, dividing along their longitudinal axis.
  • FAs initially form from multiple linear units (FAUs) of 0.3-μm width, which then separate.
  • FA-associated tension increases during splitting, and variations depend on cell type and extracellular matrix. APC and VASP depletion reduced splitting.
  • Ventral SFs appear first during cell spreading, followed by transverse arcs and dorsal SFs.

Conclusions:

  • The fundamental unit of focal adhesions is a fixed-width FAU.
  • Dynamic interactions between FAUs govern focal adhesion morphology and influence cell spreading dynamics.