December 1998
Volume 39, Issue 13
Free
Articles  |   December 1998
Use of the ARPE-19 cell line as a model of RPE polarity: basolateral secretion of FGF5.
Author Affiliations
  • K C Dunn
    Section of Molecular and Cellular Biology, University of California-Davis, USA.
  • A D Marmorstein
    Section of Molecular and Cellular Biology, University of California-Davis, USA.
  • V L Bonilha
    Section of Molecular and Cellular Biology, University of California-Davis, USA.
  • E Rodriguez-Boulan
    Section of Molecular and Cellular Biology, University of California-Davis, USA.
  • F Giordano
    Section of Molecular and Cellular Biology, University of California-Davis, USA.
  • L M Hjelmeland
    Section of Molecular and Cellular Biology, University of California-Davis, USA.
Investigative Ophthalmology & Visual Science December 1998, Vol.39, 2744-2749. doi:
  • Views
  • PDF
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      K C Dunn, A D Marmorstein, V L Bonilha, E Rodriguez-Boulan, F Giordano, L M Hjelmeland; Use of the ARPE-19 cell line as a model of RPE polarity: basolateral secretion of FGF5.. Invest. Ophthalmol. Vis. Sci. 1998;39(13):2744-2749.

      Download citation file:


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

      ×
  • Supplements
This content is PDF only. Please click on the PDF icon to access.
Abstract

PURPOSE: To determine the polarity of fibroblast growth factor 5 (FGF5) secretions from retinal pigment epithelium (RPE) cells and to examine the viability and utility of the ARPE-19 cell line as a model for the study of RPE polarity. METHODS: Influenza infection and adenovirus-mediated gene transfer were used to deliver and express genes encoding influenza hemagglutinin (HA), p75-NTR (a neurotrophin receptor), low-density lipoprotein (LDL) receptor (LDLR), and FGF5 in confluent monolayers of ARPE-19 cells. The localization of HA, p75-NTR, and LDLR was determined by confocal microscopy. Domain selective biotinylation assays were used to quantitatively determine the polarities of p75-NTR and LDLR. The secretion of FGF5 into the apical and basal media of ARPE-19 cultures was examined by immunoblot analysis of conditioned media. RESULTS: Hemagglutinin and p75-NTR were found to be localized on the apical surface of infected and transduced ARPE-19 cells. In contrast, LDLR was associated preferentially with the basolateral membrane of ARPE-19 cells. Biotinylation studies indicated that 84% of p75-NTR was present on the apical surface, and 79% of LDLR was basolaterally polarized. Over the course of 6 hours, more than 90% of the total secreted FGF5 protein accumulated in the basolateral media. CONCLUSIONS: ARPE-19 cells exhibit a polarized distribution of cell surface markers when examined by either confocal microscopy or surface-labeling assays. This indicates that the ARPE-19 cell line is a valid model for studies of RPE cell polarity. FGF5, a secreted protein normally produced by RPE cells, is accumulated preferentially in the basal media after only 6 hours, suggesting that it is vectorially secreted from the basolateral surface of ARPE-19 cells.

×
×

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.

×