Journal of Cell Science, Vol 104, Issue 2 353-363, Copyright © 1993 by Company of Biologists

JOURNAL ARTICLES

Quantitative analysis of stress fiber orientation during corneal wound contraction

WM Petroll, HD Cavanagh, P Barry, P Andrews and JV Jester
Department of Ophthalmology, University of Texas Southwestern Medical Center, Dallas 75235-9057.

Previous studies of actin and actin-binding proteins in corneal myofibroblasts suggest the development of a contractile apparatus composed, in part, of F-actin micro-filament bundles, i.e. stress fibers. To better understand the mechanics of wound contraction and the relationship between microfilament bundles and wound closure, we have analyzed the spatial and temporal organization of stress fibers during the process of corneal wound healing. Rabbit corneas (26 eyes) received 6 mm full-thickness, central incisions and were studied at various times for F-actin organization using en bloc (whole cornea) staining with FITC-phalloidin, as well as conventional histological techniques. 3-D datasets (z-series of 40 en face optical sections, 1 micron steps) were collected using the Biorad MRC-600 laser scanning confocal microscope at various regions within the wound. At 7 days, 3-D analysis showed randomly oriented, interconnected F-actin filament bundles (stress fibers). Between 7 and 28 days, stress fibers appeared to organize gradually into planes parallel to the wound surface, with a large population achieving a final orientation nearly parallel to the long axis of the wound. Using Fourier Transform analysis techniques, an orientation index (OI) was calculated to quantitate global fiber orientation at each time point. Analysis of variance demonstrated a significant change (P < 0.001) in overall stress fiber orientation from a random distribution at day 7 to an alignment more parallel to the lateral wound borders at day 28. Overall, these data suggest that stress fibers undergo temporal changes in spatial organization that correlate with wound closure, and that wound closure does not involve the development of previously described contractile or tractional forces aligned directly across the wound.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				C. Gealy, A. J. Hayes, R. Buckwell, R. D. Young, B. Caterson, A. J. Quantock, and J. R. Ralphs Actin and Type I Collagen Propeptide Distribution in the Developing Chick Cornea Invest. Ophthalmol. Vis. Sci., April 1, 2009; 50(4): 1653 - 1658. [Abstract] [Full Text] [PDF]

				M. Farid, N. Morishige, L. Lam, A. Wahlert, R. F. Steinert, and J. V. Jester Detection of Corneal Fibrosis by Imaging Second Harmonic-Generated Signals in Rabbit Corneas Treated with Mitomycin C after Excimer Laser Surface Ablation Invest. Ophthalmol. Vis. Sci., October 1, 2008; 49(10): 4377 - 4383. [Abstract] [Full Text] [PDF]

				D. Karamichos, N. Lakshman, and W. M. Petroll Regulation of Corneal Fibroblast Morphology and Collagen Reorganization by Extracellular Matrix Mechanical Properties Invest. Ophthalmol. Vis. Sci., November 1, 2007; 48(11): 5030 - 5037. [Abstract] [Full Text] [PDF]

				S. Pellegrin and H. Mellor Actin stress fibres J. Cell Sci., October 15, 2007; 120(20): 3491 - 3499. [Abstract] [Full Text] [PDF]

				V. Barbaro, E. Di Iorio, S. Ferrari, L. Bisceglia, A. Ruzza, M. De Luca, and G. Pellegrini Expression of VSX1 in Human Corneal Keratocytes during Differentiation into Myofibroblasts in Response to Wound Healing Invest. Ophthalmol. Vis. Sci., December 1, 2006; 47(12): 5243 - 5250. [Abstract] [Full Text] [PDF]

				W. M. Petroll, M. Vishwanath, and L. Ma Corneal Fibroblasts Respond Rapidly to Changes in Local Mechanical Stress Invest. Ophthalmol. Vis. Sci., October 1, 2004; 45(10): 3466 - 3474. [Abstract] [Full Text] [PDF]

				J. V. Jester, J. Huang, S. Fisher, J. Spiekerman, J. H. Chang, W. E. Wright, and J. W. Shay Myofibroblast Differentiation of Normal Human Keratocytes and hTERT, Extended-Life Human Corneal Fibroblasts Invest. Ophthalmol. Vis. Sci., May 1, 2003; 44(5): 1850 - 1858. [Abstract] [Full Text] [PDF]

				W. M. Petroll, L. Ma, and J. V. Jester Direct correlation of collagen matrix deformation with focal adhesion dynamics in living corneal fibroblasts J. Cell Sci., April 15, 2003; 116(8): 1481 - 1491. [Abstract] [Full Text] [PDF]

				M. J. CIPOLLA, N. I. GOKINA, and G. OSOL Pressure-induced actin polymerization in vascular smooth muscle as a mechanism underlying myogenic behavior FASEB J, January 1, 2002; 16(1): 72 - 76. [Abstract] [Full Text] [PDF]

				G. R. J. Melles, P. S. Binder, M. N. Moore, and J. A. Anderson Epithelial-Stromal Interactions in Human Keratotomy Wound Healing Arch Ophthalmol, September 1, 1995; 113(9): 1124 - 1130. [Abstract] [PDF]