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Related Experiment Videos

A pre-loading method of evaluating gap junctional communication by fluorescent dye transfer

G S Goldberg1, J F Bechberger, C C Naus

  • 1University of Western Ontario, London, Canada.

Biotechniques
|March 1, 1995
PubMed
Summary

This study introduces a straightforward method to assess gap junctional communication (GJC) in cultured cells using fluorescent dyes. The technique quantifies dye transfer, enabling the evaluation of GJC with potential inhibitors.

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Area of Science:

  • Cell Biology
  • Biophysics

Background:

  • Gap junctional communication (GJC) is crucial for intercellular signaling.
  • Accurate methods for quantifying GJC are essential for understanding cellular processes.

Purpose of the Study:

  • To present a simple and effective method for evaluating gap junctional communication (GJC) in cultured cells.
  • To establish a quantifiable assay for intercellular communication.

Main Methods:

  • Pre-loading cells with two fluorescent dyes: calcein (permeable through gap junctions) and DiI (not permeable).
  • Co-culturing pre-loaded cells with unlabeled cells.
  • Quantifying the number of unlabeled cells that receive calcein after settling.

Main Results:

  • The method allows for the quantification of dye transfer, indicating the extent of GJC.

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  • The system is amenable to testing potent and reversible inhibitors of GJC.

    • Conclusions:

    • This method provides a simple, quantifiable approach to assess gap junctional communication in vitro.
    • The assay can be utilized to study the effects of various compounds on GJC.