IOVS Molecular Human Reproduction
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(Investigative Ophthalmology and Visual Science. 2000;41:2357-2362.)
© 2000 by The Association for Research in Vision and Ophthalmology, Inc.

Expression and Splicing of FGF Receptor mRNAs during ARPE-19 Cell Differentiation In Vitro

Mitra Alizadeh1, Claire M. Gelfman1, Shellie R. Bench1 and Leonard M. Hjelmeland1,2

1 From the Section of Molecular and Cellular Biology, and 2 Department of Ophthalmology, University of California, Davis, California.

PURPOSE. The expression and alternative splicing of the four FGF receptor (FGFR) mRNAs are regulated in a developmental- and tissue-specific fashion. Capability of differentiation in vitro of the retinal pigment epithelial cell line ARPE-19 has been previously demonstrated. In this study, the hypothesis that FGF receptor gene expression and the alternative splicing of the FGFR1 mRNA is regulated as a function of ARPE-19 differentiation in vitro was tested.

METHODS. ARPE-19 cells were plated at sparse or confluent densities and maintained in culture up to 14 months. The expression of FGF receptors and the ratio of the FGFR1ß to FGFR1{alpha} splice variants of the FGFR1 transcript were quantified by a published PCR technique. Two in vivo samples of human RPE served as controls.

RESULTS. Sparse cultures of ARPE-19 cells predominantly express FGFR1. When these cultures are allowed to differentiate, FGFR2 is also expressed. Samples of mRNA from RPE cells in vivo exhibit FGFR1 and FGFR2 expression as well as FGFR3 expression, a form that is minimally apparent in vitro. The ratio of the FGFR1ß to FGFR1{alpha} splice variant decreases as a function of cell differentiation in vitro and approaches the ratio observed in human RPE cells in vivo. Stimulation of cultures in vitro with FGF2 as a prototypical differentiation agent does not regulate the ratio of the FGFR1ß to FGFR1{alpha} splice variant.

CONCLUSIONS. Differentiation of the ARPE-19 cell line in vitro recapitulates many but not all the in vivo patterns of FGFR expression and splicing. This in vitro system may be useful for selected studies on how cellular differentiation regulates FGF receptor gene expression and splicing.




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J.-M. Lim, S. Byun, S. Chung, T. H. Park, J.-M. Seo, C.-K. Joo, H. Chung, and D.-i. Cho
Retinal Pigment Epithelial Cell Behavior is Modulated by Alterations in Focal Cell-Substrate Contacts
Invest. Ophthalmol. Vis. Sci., November 1, 2004; 45(11): 4210 - 4216.
[Abstract] [Full Text] [PDF]

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M. Alizadeh, M. Wada, C. M. Gelfman, J. T. Handa, and L. M. Hjelmeland
Downregulation of Differentiation Specific Gene Expression by Oxidative Stress in ARPE-19 Cells
Invest. Ophthalmol. Vis. Sci., October 1, 2001; 42(11): 2706 - 2713.
[Abstract] [Full Text] [PDF]


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