Changes in cell morphology affect the quantification of intercellular communication.

ABSTRACT

The scrape loading and dye transfer (SL/DT) method is a functional assay widely used to evaluate the level of the intercellular communication via gap junction. Initially, the evaluation of the degree of communication was done by counting the rows of fluorescent cells from the scrape. Its substitution by fluorescence distance or area, determinated by morphometric softwares, was proposed as the best way of communication measurement. Considering that normal cells change their morphology during different phases of growth in monolayer cultures we proposed to evaluate the possible influence of the cell morphology changes on the quantification of intercellular communication when the SL/DT method is used. The present data indicates that GJIC capacity should be rather quantified by counting the fluorescent cell rows than by the measurement of the fluorescence distance or area, when the goal is to evaluate the cell communication along different phases of cell culture. Normal cells growing in monolayer culture decrease their diameter according to their final density of saturation, impairing the quantification of GJIC capacity by morphometric analysis.