Use of the ARPE-19 cell line as a model of RPE polarity: Basolateral secretion of FGF5

Kerrin C. Dunn, Alan D. Marmorstein, Vera L. Bonilha, Enrique Rodriguez-Boulan, Frank Giordano, Leonard M. Hjelmeland

Research output: Contribution to journalArticle

102 Scopus citations

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 anti 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.

Original languageEnglish (US)
Pages (from-to)2744-2749
Number of pages6
JournalInvestigative Ophthalmology and Visual Science
Volume39
Issue number13
StatePublished - Dec 1 1998

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

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    Dunn, K. C., Marmorstein, A. D., Bonilha, V. L., Rodriguez-Boulan, E., Giordano, F., & Hjelmeland, L. M. (1998). Use of the ARPE-19 cell line as a model of RPE polarity: Basolateral secretion of FGF5. Investigative Ophthalmology and Visual Science, 39(13), 2744-2749.