Regulation and function of extracellular matrix in intestinal epithelial restitution in vitro

Michael Göke, Anna Zuk, Daniel K. Podolsky

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81 Citations (Scopus)

Abstract

Repair of epithelial injury in the gastrointestinal tract is initially accomplished by migration of epithelial cells from the wound edge ('restitution'). To assess expression and function of the extracellular matrix (ECM) in the restitution phase after epithelial injury, in vitro studies using wounded monolayers of a rat intestinal epithelium-derived cell line (IEC-6) were undertaken. IEC-6 cells expressed fibronectin (FN) mRNA and protein in large amounts and lesser quantities of laminin-β1 (LNβ1) and LNγ1. Collagen IV (Col IV) was weakly expressed, and LNα1 was not detected. After wounding, a significant decrease in FN, LNβ1, LNγ1, and Col IVα1 mRNA steady-state levels was observed; mean content 24 h after wounding was reduced by 75-90%. FN, LN, and Col IV proteins were also reduced. The downregulation of these ECM transcripts and proteins could be substantially prevented by transforming growth factor-β1, a restitution- promoting growth factor. In addition to changes of expression, the distribution of FN and LN was also altered in migrating cells after wounding, as assessed by immunofluorescence. Arg-Gly-Asp peptides that recognize the major cell attachment site on FN and antibodies recognizing the main noncollagenous domain of Col IV inhibited cell migration, but immunoneutralizing anti-LN antisera did not affect restitution. In conclusion, although paradoxically downregulated after wounding, ECM proteins, in particular FN and Col IV molecules, are able to enhance intestinal epithelial restitution.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Gastrointestinal and Liver Physiology
Volume271
Issue number5 34-5
StatePublished - Nov 1996

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Fibronectins
Extracellular Matrix
Collagen
Extracellular Matrix Proteins
Wounds and Injuries
Down-Regulation
Messenger RNA
Transforming Growth Factors
Intestinal Mucosa
Cell Movement
Fluorescent Antibody Technique
Gastrointestinal Tract
In Vitro Techniques
Immune Sera
Intercellular Signaling Peptides and Proteins
Proteins
Epithelial Cells
laminin 1
Cell Line
Peptides

Keywords

  • basement membrane
  • cell migration
  • intestinal crypt cells
  • mucosal injury

ASJC Scopus subject areas

  • Gastroenterology
  • Physiology
  • Physiology (medical)

Cite this

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title = "Regulation and function of extracellular matrix in intestinal epithelial restitution in vitro",
abstract = "Repair of epithelial injury in the gastrointestinal tract is initially accomplished by migration of epithelial cells from the wound edge ('restitution'). To assess expression and function of the extracellular matrix (ECM) in the restitution phase after epithelial injury, in vitro studies using wounded monolayers of a rat intestinal epithelium-derived cell line (IEC-6) were undertaken. IEC-6 cells expressed fibronectin (FN) mRNA and protein in large amounts and lesser quantities of laminin-β1 (LNβ1) and LNγ1. Collagen IV (Col IV) was weakly expressed, and LNα1 was not detected. After wounding, a significant decrease in FN, LNβ1, LNγ1, and Col IVα1 mRNA steady-state levels was observed; mean content 24 h after wounding was reduced by 75-90{\%}. FN, LN, and Col IV proteins were also reduced. The downregulation of these ECM transcripts and proteins could be substantially prevented by transforming growth factor-β1, a restitution- promoting growth factor. In addition to changes of expression, the distribution of FN and LN was also altered in migrating cells after wounding, as assessed by immunofluorescence. Arg-Gly-Asp peptides that recognize the major cell attachment site on FN and antibodies recognizing the main noncollagenous domain of Col IV inhibited cell migration, but immunoneutralizing anti-LN antisera did not affect restitution. In conclusion, although paradoxically downregulated after wounding, ECM proteins, in particular FN and Col IV molecules, are able to enhance intestinal epithelial restitution.",
keywords = "basement membrane, cell migration, intestinal crypt cells, mucosal injury",
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T1 - Regulation and function of extracellular matrix in intestinal epithelial restitution in vitro

AU - Göke, Michael

AU - Zuk, Anna

AU - Podolsky, Daniel K.

PY - 1996/11

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N2 - Repair of epithelial injury in the gastrointestinal tract is initially accomplished by migration of epithelial cells from the wound edge ('restitution'). To assess expression and function of the extracellular matrix (ECM) in the restitution phase after epithelial injury, in vitro studies using wounded monolayers of a rat intestinal epithelium-derived cell line (IEC-6) were undertaken. IEC-6 cells expressed fibronectin (FN) mRNA and protein in large amounts and lesser quantities of laminin-β1 (LNβ1) and LNγ1. Collagen IV (Col IV) was weakly expressed, and LNα1 was not detected. After wounding, a significant decrease in FN, LNβ1, LNγ1, and Col IVα1 mRNA steady-state levels was observed; mean content 24 h after wounding was reduced by 75-90%. FN, LN, and Col IV proteins were also reduced. The downregulation of these ECM transcripts and proteins could be substantially prevented by transforming growth factor-β1, a restitution- promoting growth factor. In addition to changes of expression, the distribution of FN and LN was also altered in migrating cells after wounding, as assessed by immunofluorescence. Arg-Gly-Asp peptides that recognize the major cell attachment site on FN and antibodies recognizing the main noncollagenous domain of Col IV inhibited cell migration, but immunoneutralizing anti-LN antisera did not affect restitution. In conclusion, although paradoxically downregulated after wounding, ECM proteins, in particular FN and Col IV molecules, are able to enhance intestinal epithelial restitution.

AB - Repair of epithelial injury in the gastrointestinal tract is initially accomplished by migration of epithelial cells from the wound edge ('restitution'). To assess expression and function of the extracellular matrix (ECM) in the restitution phase after epithelial injury, in vitro studies using wounded monolayers of a rat intestinal epithelium-derived cell line (IEC-6) were undertaken. IEC-6 cells expressed fibronectin (FN) mRNA and protein in large amounts and lesser quantities of laminin-β1 (LNβ1) and LNγ1. Collagen IV (Col IV) was weakly expressed, and LNα1 was not detected. After wounding, a significant decrease in FN, LNβ1, LNγ1, and Col IVα1 mRNA steady-state levels was observed; mean content 24 h after wounding was reduced by 75-90%. FN, LN, and Col IV proteins were also reduced. The downregulation of these ECM transcripts and proteins could be substantially prevented by transforming growth factor-β1, a restitution- promoting growth factor. In addition to changes of expression, the distribution of FN and LN was also altered in migrating cells after wounding, as assessed by immunofluorescence. Arg-Gly-Asp peptides that recognize the major cell attachment site on FN and antibodies recognizing the main noncollagenous domain of Col IV inhibited cell migration, but immunoneutralizing anti-LN antisera did not affect restitution. In conclusion, although paradoxically downregulated after wounding, ECM proteins, in particular FN and Col IV molecules, are able to enhance intestinal epithelial restitution.

KW - basement membrane

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