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Visualizing Spatiotemporal Dynamics of Intercellular Mechanotransmission upon Wounding.

Pengzhi Wang1,2, Jing Liang3, Linda Z Shi2

  • 1Department of Bioengineering, University of California, San Diego, La Jolla, California 92093, United States.

ACS Photonics
|May 10, 2019
PubMed
Summary

Researchers developed a new method combining laser microscissors and FRET microscopy to visualize cell signaling. This technique revealed that cell-contact loss triggers Src kinase activation dependent on cytoskeleton, not actomyosin contractility.

Keywords:
FRET imagingactive actomyosin contractilitydirected evolutionhigh-throughput screeninghighly sensitive FRET-based biosensor SCAGElaser-induced woundingpassive structural support of cytoskeletontransient Src activation

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Area of Science:

  • Cell Biology
  • Biophysics
  • Molecular Imaging

Background:

  • Intercellular communication relies on physical and biochemical cues, crucial for multicellular organism function.
  • Visualizing dynamic intercellular signaling in single live cells remains a significant challenge.
  • Understanding the interplay between physical contact and biochemical signaling is vital.

Purpose of the Study:

  • To develop a photonic approach for visualizing intercellular signaling dynamics in live cells.
  • To investigate the role of physical cues in triggering biochemical signaling cascades.
  • To study the molecular mechanisms underlying cell-cell contact loss and subsequent signaling events.

Main Methods:

  • Development of a highly sensitive Förster resonance energy transfer (FRET)-based biosensor (SCAGE) for Src kinase.
  • Utilized femtosecond laser pulses ('laser microscissors') to precisely sever physical intercellular connections.
  • Employed FRET microscopy to visualize real-time Src kinase activation in response to physical perturbations.

Main Results:

  • The novel SCAGE biosensor demonstrated over 40-fold sensitivity enhancement in live mammalian cells.
  • Severing cell-cell contacts induced a transient Src kinase activation in neighboring cells.
  • This Src activation was dependent on passive cytoskeletal support, independent of active actomyosin contractility.

Conclusions:

  • The integrated photonic approach enables precise physical perturbation and visualization of intercellular signaling.
  • This method provides a unique system for studying the physical-biochemical basis of intercellular coupling and wounding.
  • Findings elucidate the role of the cytoskeleton in mediating signaling responses to mechanical stress.