Analysis of the pattern of subcellular force generation by corneal fibroblasts after Rho activation

Eye & Contact Lens
W Matthew PetrollMridula Vishwanath

Abstract

To determine the structural and subcellular mechanical effects of Rho activation on corneal fibroblasts in three-dimensional collagen matrices. Human corneal fibroblasts were plated at low density in 100-microm thick fibrillar collagen matrices and cultured for 1 or 2 days in serum-free media. Time-lapse imaging was then performed at 1- to 2-minute intervals with Nomarski differential interference contrast. After 1 hour, perfusion was switched to serum-free media containing 1 micromol/L lysophosphatidic acid (LPA). After an additional 30 to 60 minutes, the Rho kinase (ROCK) inhibitor Y-27632 was added to the perfusion media. Changes in cell structure and extracellular matrix deformation were measured with MetaMorph. Addition of LPA activated Rho and induced retraction of cell processes and cellular contraction, as indicated by decreases in cell length (-12.1%+/-7.0%; P<0.05) and cell area (-13.1%+/-13.5%; P=0.06). Force generation was greatest along the cell body in all cases, as indicated by the location of maximum extracellular matrix compression. Subsequent addition of Y-27632 resulted in relaxation of extracellular matrix stress, and reextension of cellular processes. The data show that Rho induces rapid contraction of corn...Continue Reading

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Citations

Jul 7, 2009·Annual Review of Cell and Developmental Biology·Frederick Grinnell, W Matthew Petroll
Dec 23, 2009·Experimental Eye Research·Neema LakshmanW Matthew Petroll
Dec 9, 2008·Cell Motility and the Cytoskeleton·Dimitris KaramichosW Matthew Petroll
Mar 31, 2015·Experimental Eye Research·W Matthew Petroll, Miguel Miron-Mendoza
Jul 4, 2013·Experimental Cell Research·Miguel Miron-MendozaW Matthew Petroll
Nov 26, 2013·Biophysical Journal·Karin A JansenGijsje H Koenderink
Dec 7, 2010·Cellular and Molecular Bioengineering·Chengxin Zhou, W Matthew Petroll

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