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Changes in cell morphology affect the quantification of intercellular communication.

Beatriz Brandão Vaz-de-Lima1, Marisa Ionta, Glaucia M Machado-Santelli

  • 1Universidade de São Paulo, Instituto de Ciências Biomédicas, Departamento de Biologia Celular e do Desenvolvimento, Sao Paulo, Brazil.

Micron (Oxford, England : 1993)
|June 6, 2007
PubMed
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Cell morphology changes impact gap junction intercellular communication (GJIC) measurements. Counting fluorescent cell rows is more reliable than morphometric analysis for evaluating GJIC across cell culture phases.

Area of Science:

  • Cell Biology
  • Biophysics

Background:

  • Gap junction intercellular communication (GJIC) is vital for cellular coordination.
  • The scrape loading and dye transfer (SL/DT) assay is a common method to measure GJIC.
  • Morphometric analysis using fluorescence distance or area has been proposed to improve SL/DT quantification.

Purpose of the Study:

  • To investigate the influence of cell morphology changes on GJIC quantification using the SL/DT method.
  • To determine the most reliable method for measuring GJIC in cells at different growth phases.

Main Methods:

  • Utilized the scrape loading and dye transfer (SL/DT) assay.
  • Compared quantification by counting fluorescent cell rows versus morphometric analysis (fluorescence distance/area).
  • Observed normal cells in monolayer cultures across different growth phases.

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Main Results:

  • Cellular diameter decreases as cells reach saturation density in monolayer cultures.
  • Morphometric quantification of GJIC (distance/area) is impaired by these cell morphology changes.
  • Counting fluorescent cell rows provides a more consistent measure of GJIC capacity.

Conclusions:

  • Cell morphology variations significantly affect morphometric quantification of GJIC.
  • Counting fluorescent cell rows is the preferred method for reliable GJIC assessment across different cell culture stages.
  • Avoid morphometric analysis for GJIC when cell morphology is variable.