Oxidant and antioxidant modulation of chloride channels expressed in human retinal pigment epithelium

T. X. Weng, B. F. Godley, G. F. Jin, N. J. Mangini, B. G. Kennedy, A. S.L. Yu, N. K. Wills

Research output: Contribution to journalArticle

42 Scopus citations

Abstract

Retinal pigment epithelium (RPE) possesses regulated chloride channels that are crucial for transepithelial fluid and ion transport. At present, little is known about the molecular nature of chloride channels in human adult RPE (haRPE) or the effects of oxidative stress on membrane conductance properties. In the present study, we assessed ClC channel and cystic fibrosis transmembrane conductance regulator (CFTR) expression and membrane chloride conductance properties in haRPE cells. ClC-5, ClC-3, ClC-2, and CFTR mRNA expression was confirmed with RT-PCR analysis, and protein expression was detected with Western blot analysis and immunofluorescence microscopy. Whole cell recordings of primary cultures of haRPE showed an outwardly rectifying chloride current that was inhibited by the oxidant H 2O 2. The inhibitory effects of H 2O 2 were reduced in cultured human RPE cells that were incubated with precursors of glutathione synthesis or that were stably transfected to overexpress glutathione S-transferase. These findings indicate a possible role for ClC channels in haRPE cells and suggest possible redox modulation of human RPE chloride conductances.

Original languageEnglish (US)
Pages (from-to)C839-C849
JournalAmerican Journal of Physiology - Cell Physiology
Volume283
Issue number3 52-3
StatePublished - Sep 1 2002

Keywords

  • Glutathione
  • Glutathione S-transferase
  • Immunocytochemistry
  • Patch clamp

ASJC Scopus subject areas

  • Physiology
  • Cell Biology

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    Weng, T. X., Godley, B. F., Jin, G. F., Mangini, N. J., Kennedy, B. G., Yu, A. S. L., & Wills, N. K. (2002). Oxidant and antioxidant modulation of chloride channels expressed in human retinal pigment epithelium. American Journal of Physiology - Cell Physiology, 283(3 52-3), C839-C849.