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Biophys. J. BioFAST: First Published May 27, 2005. doi:10.1529/biophysj.105.061218
© 2005 by the Biophysical Society.

A more recent version of this article appeared on August 1, 2005.
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CELL BIOPHYSICS

Spatial Analysis of 3' Phosphoinositide Signaling in Living Fibroblasts: III. Influence of Cell Morphology and Morphological Polarity

Ian C. Schneider 1, Elizabeth M. Parrish 1 and Jason M. Haugh 1*

1 North Carolina State University

* To whom correspondence should be addressed. E-mail: jason_haugh{at}ncsu.edu.

Submitted on February 15, 2005
Revised on March 27, 2005
Accepted on 23 May 2005


  Abstract
Activation of phosphoinositide (PI) 3-kinase is a required signaling pathway in fibroblast migration directed by platelet-derived growth factor (PDGF). The pattern of 3' PI lipids in the plasma membrane, integrating local PI 3-kinase activity as well as 3' PI diffusion and turnover, influences the spatiotemporal regulation of the cytoskeleton. In fibroblasts stimulated uniformly with PDGF, visualized using total internal reflection fluorescence (TIRF) microscopy, we consistently observed localized regions with significantly higher or lower 3' PI levels than adjacent regions (hot and cold spots, respectively). A typical cell contained multiple hot spots, coinciding with apparent leading edge structures, and at most one cold spot at the rear. Using a framework for finite-element modeling with actual cell contact area geometries, we find that although the 3' PI pattern is affected by irregular contact area shape, cell morphology alone cannot explain the presence of hot or cold spots. Our results and analysis instead suggest that these regions reflect different local 3' PI dynamics, specifically through a combination of mechanisms: enhanced PI 3-kinase activity, reduced 3' PI turnover, and possibly slow/constrained 3' PI diffusion. The morphological polarity of the cell may thus bias 3' PI signaling to promote persistent migration in fibroblasts.

Key Words: diffusion, fluorescence, gradient, mathematical model, phosphatidylinositol 3-kinase, signal transduction




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Copyright © 2005 by the Biophysical Society.