Visualizing Spatiotemporal Dynamics of Intercellular Mechanotransmission upon Wounding

Pengzhi Wang, Jing Liang, Linda Z. Shi, Yi Wang, Ping Zhang, Mingxing Ouyang, Daryl Preece, Qin Peng, Lunan Shao, Jason Fan, Jie Sun, Shawn S. Li, Michael W. Berns, Huimin Zhao, Yingxiao Wang

Research output: Contribution to journalArticle

Abstract

During cell-to-cell communications, the interplay between physical and biochemical cues is essential for informational exchange and functional coordination, especially in multicellular organisms. However, it remains a challenge to visualize intercellular signaling dynamics in single live cells. Here, we report a photonic approach, based on laser microscissors and Förster resonance energy transfer (FRET) microscopy, to study intercellular signaling transmission. First, using our high-throughput screening platform, we developed a highly sensitive FRET-based biosensor (SCAGE) for Src kinase, a key regulator of intercellular interactions and signaling cascades. Notably, SCAGE showed a more than 40-fold sensitivity enhancement than the original biosensor in live mammalian cells. Next, upon local severance of physical intercellular connections by femtosecond laser pulses, SCAGE enabled the visualization of a transient Src activation across neighboring cells. Lastly, we found that this observed transient Src activation following the loss of cell-cell contacts depends on the passive structural support of cytoskeleton but not on the active actomyosin contractility. Hence, by precisely introducing local physical perturbations and directly visualizing spatiotemporal transmission of ensuing signaling events, our integrated approach could be broadly applied to mimic and investigate the wounding process at single-cell resolutions. This integrated approach with highly sensitive FRET-based biosensors provides a unique system to advance our in-depth understanding of molecular mechanisms underlying the physical-biochemical basis of intercellular coupling and wounding processes.

Original languageEnglish (US)
Pages (from-to)3565-3574
Number of pages10
JournalACS Photonics
Volume5
Issue number9
DOIs
StatePublished - Sep 19 2018

Keywords

  • FRET imaging
  • active actomyosin contractility
  • directed evolution
  • high-throughput screening
  • highly sensitive FRET-based biosensor SCAGE
  • laser-induced wounding
  • passive structural support of cytoskeleton
  • transient Src activation

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Biotechnology
  • Atomic and Molecular Physics, and Optics
  • Electrical and Electronic Engineering

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