December 2002
Volume 43, Issue 13
Free
ARVO Annual Meeting Abstract  |   December 2002
Epidermal Growth Factor and Endothelin-1 Regulation of Mitogen-Activated Protein Kinase Activity, Na-K-2Cl Cotransporter and Na/K-ATPase Expression and Proliferation in Bovine Corneal Epithelial Cells
Author Affiliations & Notes
  • VN Bildin
    Biological Sciences SUNY College of Optometry New York NY
  • Z Wang
    New York NY
  • PS Reinach
    New York NY
  • Footnotes
    Commercial Relationships   V.N. Bildin, None; Z. Wang , None; P.S. Reinach , None. Grant Identification: EY04795 and the SIVR Foundation
Investigative Ophthalmology & Visual Science December 2002, Vol.43, 1651. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      VN Bildin, Z Wang, PS Reinach; Epidermal Growth Factor and Endothelin-1 Regulation of Mitogen-Activated Protein Kinase Activity, Na-K-2Cl Cotransporter and Na/K-ATPase Expression and Proliferation in Bovine Corneal Epithelial Cells . Invest. Ophthalmol. Vis. Sci. 2002;43(13):1651.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Abstract: : Purpose: To determine if there is an interaction between epidermal growth factor (EGF) and endothelin-1 (ET-1) in their stimulation of mitogen-activated protein kinase (MAPK) activity, Na-K-2Cl cotransporter (NKCC) and Na/K-ATPase protein expression as well as proliferation in cultured bovine corneal epithelial cells (BCEC). Methods: After 48 h of serum starvation, the BCEC were incubated with or without EGF and/or ET-1 at different concentrations: 1.) for 10 min to analyze MAPK activation; 2.) 24 h to determine subsequent changes in NKCC and Na/K-ATPase protein expression and cell proliferation. Western blot/ECL assay was used to evaluate phosphorylated MAPK, NKCC and Na/K-ATPase protein amounts in the cytosolic fraction and membrane-containing pellet of cell lysates, respectively. Proliferation was quantified based on measurements of 3H-thymidine uptake. Results: Both mitogens applied alone dose dependently stimulated MAPK activity with a maximal increase of about 10-fold for 1-10 nM ET-1 and more than 20-fold for 5-10 ng/ml EGF. EGF and ET-1 at those concentrations similarly upregulated NKCC and Na/K-ATPase proteins expression levels, (i.e. 6-7- and 3-4-fold, respectively). Cell proliferation was also proportionally stimulated. On the other hand, the combined effects of EGF and ET-1 at all concentrations on MAPK activity, NKCC and Na/K-ATPase proteins levels and cell proliferation were similar to those with 5-10 ng/ml EGF alone. Conclusion: The dose dependent stimulation of NKCC and Na/K-ATPase protein expression and proliferation induced by EGF and ET-1 in BCEC closely correlate with increases in MAPK activity produced by these cytokines. This suggests that EGF and ET-1 receptor control of proliferation occurs through changes in cell volume. EGF and ET-1 receptor regulation of NKCC and Na/K-ATPase protein expression and cell proliferation both occur through common signaling pathways and involve MAPK activation.

Keywords: 423 growth factors/growth factor receptors • 445 ion channels • 372 cornea: epithelium 
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×