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Imaging dynamic cell-cell junctional coupling in vivo using Trojan-LAMP.

Yan-Ming Guo1, Shiuhwei Chen, Premnath Shetty

  • 1Department of Cell Biology, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, Texas 75390, USA.

Nature Methods
|January 23, 2009
PubMed
Summary

Researchers developed a new fluorescent probe, dextran-CANPE-HCC, to visualize cell-cell gap junction communication in vivo. This method revealed dynamic remodeling of cell coupling patterns during early embryonic development in Caenorhabditis elegans.

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Area of Science:

  • Developmental Biology
  • Cell Biology
  • Biochemistry

Background:

  • Cell-cell communication via gap junctions is crucial for physiological regulation and function.
  • Existing methods for imaging gap junction communication in vivo have limitations.
  • A novel imaging tool is needed to study dynamic intercellular communication in developing organisms.

Purpose of the Study:

  • To develop and validate a novel bioconjugate for imaging cell coupling in vivo.
  • To investigate the dynamics of gap junction communication during early embryonic development.
  • To establish a new technique for noninvasive imaging of cell coupling in living organisms.

Main Methods:

  • Development of dextran-CANPE-HCC, a caged fluorescent dye bioconjugate.
  • In vitro characterization of photolysis efficiency and fluorescence enhancement.

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  • Application of Trojan-local activation of molecular fluorescent probes (LAMP) in Caenorhabditis elegans.
  • In vivo imaging of intercellular dye transfer through gap junction channels.

    • Main Results:

    • Dextran-CANPE-HCC demonstrated efficient photolysis and robust fluorescence enhancement in vitro.
    • The bioconjugate was successfully delivered and retained in Caenorhabditis elegans oocytes throughout development.
    • LAMP imaging revealed dynamic remodeling of cell coupling patterns during embryogenesis, isolating germ cell precursors.
    • Stable labeling allowed noninvasive examination of cell coupling in hatched worms.

    Conclusions:

    • Dextran-CANPE-HCC and the LAMP technique provide a powerful new tool for studying in vivo cell coupling.
    • The study elucidated dynamic changes in gap junction communication during early embryonic development.
    • This approach enables noninvasive, high-resolution imaging of intercellular communication in live model organisms.