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. [3H] 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 language | English (US) |
---|---|
Pages (from-to) | 2614-2622 |
Number of pages | 9 |
Journal | Investigative Ophthalmology and Visual Science |
Volume | 36 |
Issue number | 13 |
State | Published - 1995 |
Keywords
- cornea
- cytokines
- endothelin
- epithelium
- gene expression
ASJC Scopus subject areas
- Ophthalmology
- Sensory Systems
- Cellular and Molecular Neuroscience