Quantitative analysis of stress fiber orientation during corneal wound contraction

Walter M Petroll, Harrison D Cavanagh, Patricia Barry, Peter Andrews, James V. Jester

Research output: Contribution to journalArticle

91 Citations (Scopus)

Abstract

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 μm 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.

Original languageEnglish (US)
Pages (from-to)353-363
Number of pages11
JournalJournal of Cell Science
Volume104
Issue number2
StatePublished - Feb 1993

Fingerprint

Stress Fibers
Wounds and Injuries
Actins
Actin Cytoskeleton
Fourier Analysis
Cornea
Histological Techniques
Phalloidine
Microfilament Proteins
Myofibroblasts
Fluorescein-5-isothiocyanate
Mechanics
Wound Healing
Analysis of Variance
Lasers
Staining and Labeling
Rabbits

Keywords

  • Confocal microscopy
  • Myofibroblasts
  • Stress fibres
  • Wound contraction

ASJC Scopus subject areas

  • Cell Biology

Cite this

Quantitative analysis of stress fiber orientation during corneal wound contraction. / Petroll, Walter M; Cavanagh, Harrison D; Barry, Patricia; Andrews, Peter; Jester, James V.

In: Journal of Cell Science, Vol. 104, No. 2, 02.1993, p. 353-363.

Research output: Contribution to journalArticle

Petroll, Walter M ; Cavanagh, Harrison D ; Barry, Patricia ; Andrews, Peter ; Jester, James V. / Quantitative analysis of stress fiber orientation during corneal wound contraction. In: Journal of Cell Science. 1993 ; Vol. 104, No. 2. pp. 353-363.
@article{f2eaacfb822e4a5b9ba1381d4f9ff08a,
title = "Quantitative analysis of stress fiber orientation during corneal wound contraction",
abstract = "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 μm 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.",
keywords = "Confocal microscopy, Myofibroblasts, Stress fibres, Wound contraction",
author = "Petroll, {Walter M} and Cavanagh, {Harrison D} and Patricia Barry and Peter Andrews and Jester, {James V.}",
year = "1993",
month = "2",
language = "English (US)",
volume = "104",
pages = "353--363",
journal = "Journal of Cell Science",
issn = "0021-9533",
publisher = "Company of Biologists Ltd",
number = "2",

}

TY - JOUR

T1 - Quantitative analysis of stress fiber orientation during corneal wound contraction

AU - Petroll, Walter M

AU - Cavanagh, Harrison D

AU - Barry, Patricia

AU - Andrews, Peter

AU - Jester, James V.

PY - 1993/2

Y1 - 1993/2

N2 - 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 μm 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.

AB - 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 μm 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.

KW - Confocal microscopy

KW - Myofibroblasts

KW - Stress fibres

KW - Wound contraction

UR - http://www.scopus.com/inward/record.url?scp=0027502812&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027502812&partnerID=8YFLogxK

M3 - Article

VL - 104

SP - 353

EP - 363

JO - Journal of Cell Science

JF - Journal of Cell Science

SN - 0021-9533

IS - 2

ER -