Cytoskeleton Modulation — Chemical Screen


Paran et al., J. Struct. Biol. 158 (2007) 233–243 

 

Chemical modulators of focal adhesions (FA) were screened using fluorescently labeled fibroblasts. Effects were automatically detected using multiparametric evaluation of FA morphological and intensity features in control versus treated wells. This methodology was used to measure dose-dependent and time-dependent effects of the compounds.

Fibroblasts expressing fluorescently labeled Paxillin, a protein localized at the focal adhesion complex, were treated with chemicals and natural extracts. Watershed segmentation was used to define the focal adhesions. Quantification of morphological and intensity features of the cells and their focal adhesions were used to automatically define affected cells (right and middle) in comparison to control (left). Scale bar - 20 ?m.


Dose-dependent effect of a crude natural extract from the marine sponge, Jaspis sp. (A) Typical images of cells treated with extract; 100 ?g/ml causes massive cell damage 1 ?g/ml induces doublets of small FAs, while FAs remain similar to control at 10 ng/ml. Scale bar: 20 ?m. The lower panel in (A) shows quantitative analysis of FA area using Kolmogorov–Smirnov statistics (KSsa). The colors mark the scores; red denotes a large deviation from control and green indicates similarity to control. Each dilution was tested in well duplicate (~60 cells). (B) An example of FA area histograms that were used to calculate the KSsa factor between control and treated cells.