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Related Experiment Video

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Genetically Encoded Fluorescent Indicator GRAPHIC Delineates Intercellular Connections.

Nagatoki Kinoshita1, Arthur J Y Huang2, Thomas J McHugh2

  • 1Molecular Mechanisms of Brain Development, Center for Brain Science (CBS), RIKEN, Saitama, Japan; Exploratory Research for Advanced Technology (ERATO), Japan Science and Technology Agency (JST), Tokyo, Japan.

Iscience
|April 27, 2019
PubMed
Summary

Researchers developed GRAPHIC, a new fluorescent tool to visualize cell-cell connections. This technology precisely maps intercellular contacts in various tissues, aiding the study of complex multicellular systems.

Keywords:
Biological SciencesCell BiologyMolecular Biology

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Area of Science:

  • Cell Biology
  • Neuroscience
  • Developmental Biology

Background:

  • Intercellular contacts are crucial for organ development and function.
  • Understanding the precise location and timing of cell-cell adhesions is challenging in many biological systems.

Purpose of the Study:

  • To develop a novel genetically encoded fluorescent indicator for visualizing intercellular contacts.
  • To demonstrate the utility of this indicator in various biological contexts, including cell culture, fish retina, and mouse brain.

Main Methods:

  • Development of GRAPHIC (GPI anchored reconstitution-activated proteins highlight intercellular connections), a fluorescent indicator utilizing optimized intercellular GFP reconstitution via a GPI anchor.
  • Application of GRAPHIC in cultured cells, zebrafish retina, and mouse central nervous system.
  • Generation of GRAPHIC color variants for simultaneous detection of multiple contacts.

Main Results:

  • GRAPHIC successfully visualized intercellular contacts at the interface of cultured cells without disrupting natural adhesion.
  • The tool delineated specific cone-bipolar connection sites in the fish retina.
  • GRAPHIC identified synaptic sites within the mouse thalamocortical circuit.
  • Color variants enabled simultaneous detection of multiple convergent contacts in cell culture.

Conclusions:

  • GRAPHIC is a sensitive and versatile tool for analyzing complex multicellular connections.
  • This technology overcomes previous limitations in visualizing spatiotemporal information of cell-cell contacts.
  • GRAPHIC facilitates the study of organogenesis, function, and maintenance by providing detailed insights into intercellular interactions.