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Mapping Mechanical Strain of an Endogenous Cytoskeletal Network in Living Endothelial Cells

Brian P. Helmke^*, Amy B. Rosen^, and Peter F. Davies^,,

^* Department of Biomedical Engineering and Cardiovascular Research Center, University of Virginia, Charlottesville, Virginia; and Institute for Medicine and Engineering, Department of Bioengineering, and Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania

Correspondence: Address reprint requests to Peter F. Davies, Institute for Medicine and Engineering, University of Pennsylvania, 1010 Vagelos Research Laboratories, 3340 Smith Walk, Philadelphia, PA 19104-6383. Tel.: 215-573-6813; Fax: 215-573-6815; E-mail: pfd{at}pobox.upenn.edu.

A central aspect of cellular mechanochemical signaling is a change of cytoskeletal tension upon the imposition of exogenous forces. Here we report measurements of the spatiotemporal distribution of mechanical strain in the intermediate filament cytoskeleton of endothelial cells computed from the relative displacement of endogenous green fluorescent protein (GFP)-vimentin before and after onset of shear stress. Quantitative image analysis permitted computation of the principal values and orientations of Lagrangian strain from 3-D high-resolution fluorescence intensity distributions that described intermediate filament positions. Spatially localized peaks in intermediate filament strain were repositioned after onset of shear stress. The orientation of principal strain indicated that mechanical stretching was induced across cell boundaries. This novel approach for intracellular strain mapping using an endogenous reporter demonstrates force transfer from the lumenal surface throughout the cell.

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