ET(B) and epidermal growth factor receptor stimulation of wound closure in bovine corneal epithelial cells

W. Tao, Gregory I Liou, X. Wu, P. S. Reinach

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Purpose. To determine if there is a heterogeneous pattern of endothelin (ET) receptor subtype (i.e., ET(A) and ET(B)) gene expression in the bovine corneal epithelium (BCE). To determine it ET receptor subtype stimulation increases the effectiveness of epidermal growth factor (EGF) to accelerate wound closure in a primary culture of bovine corneal epithelial cells (BCEC). Methods. In situ hybridization histochemistry was used to characterize ET(A) and ET(B) gene expression in the BCE. A wound closure assay evaluated wound healing rates in BCEC after 4 to 7 days in culture. [ 3 H] thymidine incorporation and MTT assay measured proliferation. Results. ET(A) gene expression was appreciably higher in the basal cells than in the suprabasal cells, whereas the pattern for ET(B) was reversed. Epidermal growth factor (5 ng/ml) maximally increased wound closure by 145% above the control. With 5 ng/ml EGF, either 10 -9 M ET-1 or 10 -8 M sarafotoxin-6-c (s-6-c) increased wound closure by an additional 39% (P < 0.001) above that measured with 5 ng/ml EGF alone. BQ123 (10 -7 M) did not alter any of these effects of ET-1 or s-6-c. Epidermal growth factor stimulated wound closure through a selective increase in proliferation. Neither ET-1 nor s-6-c alone had any effect on proliferation or migration. Conclusions. Both ET(A) and .ET(B) genes are expressed in BCE. However, in BCEC only, ET(B) stimulation increases the effectiveness of EGF to stimulate wound closure. This response was caused by an increase in cell migration rather than proliferation because, after treatment with mitomycin C, neither ET-1 nor EGF stimulated wound closure.

Original languageEnglish (US)
Pages (from-to)2614-2622
Number of pages9
JournalInvestigative Ophthalmology and Visual Science
Volume36
Issue number13
StatePublished - Jan 1 1995

Fingerprint

Endothelins
Epidermal Growth Factor Receptor
Epidermal Growth Factor
Epithelial Cells
Wounds and Injuries
Endothelin-1
Corneal Epithelium
Endothelin Receptors
Gene Expression
Mitomycin
Wound Healing
Thymidine
Cell Movement
In Situ Hybridization

Keywords

  • cornea
  • cytokines
  • endothelin
  • epithelium
  • gene expression

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

ET(B) and epidermal growth factor receptor stimulation of wound closure in bovine corneal epithelial cells. / Tao, W.; Liou, Gregory I; Wu, X.; Reinach, P. S.

In: Investigative Ophthalmology and Visual Science, Vol. 36, No. 13, 01.01.1995, p. 2614-2622.

Research output: Contribution to journalArticle

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abstract = "Purpose. To determine if there is a heterogeneous pattern of endothelin (ET) receptor subtype (i.e., ET(A) and ET(B)) gene expression in the bovine corneal epithelium (BCE). To determine it ET receptor subtype stimulation increases the effectiveness of epidermal growth factor (EGF) to accelerate wound closure in a primary culture of bovine corneal epithelial cells (BCEC). Methods. In situ hybridization histochemistry was used to characterize ET(A) and ET(B) gene expression in the BCE. A wound closure assay evaluated wound healing rates in BCEC after 4 to 7 days in culture. [ 3 H] thymidine incorporation and MTT assay measured proliferation. Results. ET(A) gene expression was appreciably higher in the basal cells than in the suprabasal cells, whereas the pattern for ET(B) was reversed. Epidermal growth factor (5 ng/ml) maximally increased wound closure by 145{\%} above the control. With 5 ng/ml EGF, either 10 -9 M ET-1 or 10 -8 M sarafotoxin-6-c (s-6-c) increased wound closure by an additional 39{\%} (P < 0.001) above that measured with 5 ng/ml EGF alone. BQ123 (10 -7 M) did not alter any of these effects of ET-1 or s-6-c. Epidermal growth factor stimulated wound closure through a selective increase in proliferation. Neither ET-1 nor s-6-c alone had any effect on proliferation or migration. Conclusions. Both ET(A) and .ET(B) genes are expressed in BCE. However, in BCEC only, ET(B) stimulation increases the effectiveness of EGF to stimulate wound closure. This response was caused by an increase in cell migration rather than proliferation because, after treatment with mitomycin C, neither ET-1 nor EGF stimulated wound closure.",
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AU - Liou, Gregory I

AU - Wu, X.

AU - Reinach, P. S.

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N2 - Purpose. To determine if there is a heterogeneous pattern of endothelin (ET) receptor subtype (i.e., ET(A) and ET(B)) gene expression in the bovine corneal epithelium (BCE). To determine it ET receptor subtype stimulation increases the effectiveness of epidermal growth factor (EGF) to accelerate wound closure in a primary culture of bovine corneal epithelial cells (BCEC). Methods. In situ hybridization histochemistry was used to characterize ET(A) and ET(B) gene expression in the BCE. A wound closure assay evaluated wound healing rates in BCEC after 4 to 7 days in culture. [ 3 H] thymidine incorporation and MTT assay measured proliferation. Results. ET(A) gene expression was appreciably higher in the basal cells than in the suprabasal cells, whereas the pattern for ET(B) was reversed. Epidermal growth factor (5 ng/ml) maximally increased wound closure by 145% above the control. With 5 ng/ml EGF, either 10 -9 M ET-1 or 10 -8 M sarafotoxin-6-c (s-6-c) increased wound closure by an additional 39% (P < 0.001) above that measured with 5 ng/ml EGF alone. BQ123 (10 -7 M) did not alter any of these effects of ET-1 or s-6-c. Epidermal growth factor stimulated wound closure through a selective increase in proliferation. Neither ET-1 nor s-6-c alone had any effect on proliferation or migration. Conclusions. Both ET(A) and .ET(B) genes are expressed in BCE. However, in BCEC only, ET(B) stimulation increases the effectiveness of EGF to stimulate wound closure. This response was caused by an increase in cell migration rather than proliferation because, after treatment with mitomycin C, neither ET-1 nor EGF stimulated wound closure.

AB - Purpose. To determine if there is a heterogeneous pattern of endothelin (ET) receptor subtype (i.e., ET(A) and ET(B)) gene expression in the bovine corneal epithelium (BCE). To determine it ET receptor subtype stimulation increases the effectiveness of epidermal growth factor (EGF) to accelerate wound closure in a primary culture of bovine corneal epithelial cells (BCEC). Methods. In situ hybridization histochemistry was used to characterize ET(A) and ET(B) gene expression in the BCE. A wound closure assay evaluated wound healing rates in BCEC after 4 to 7 days in culture. [ 3 H] thymidine incorporation and MTT assay measured proliferation. Results. ET(A) gene expression was appreciably higher in the basal cells than in the suprabasal cells, whereas the pattern for ET(B) was reversed. Epidermal growth factor (5 ng/ml) maximally increased wound closure by 145% above the control. With 5 ng/ml EGF, either 10 -9 M ET-1 or 10 -8 M sarafotoxin-6-c (s-6-c) increased wound closure by an additional 39% (P < 0.001) above that measured with 5 ng/ml EGF alone. BQ123 (10 -7 M) did not alter any of these effects of ET-1 or s-6-c. Epidermal growth factor stimulated wound closure through a selective increase in proliferation. Neither ET-1 nor s-6-c alone had any effect on proliferation or migration. Conclusions. Both ET(A) and .ET(B) genes are expressed in BCE. However, in BCEC only, ET(B) stimulation increases the effectiveness of EGF to stimulate wound closure. This response was caused by an increase in cell migration rather than proliferation because, after treatment with mitomycin C, neither ET-1 nor EGF stimulated wound closure.

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