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Expression and splicing of FGF receptor mRNAs during APRE-19 cell differentiation in vitro
M Alizadeh1, C M Gelfman, S R Bench
1Section of Molecular and Cellular Biology, University of California, Davis 95616-8794, USA.
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 FGFR1beta to FGFR1alpha 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 FGFR1beta to FGFR1alpha 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 FGFR1beta to FGFR1alpha 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.
Insights
Cellular differentiation in vitro affects Fibroblast Growth Factor Receptor (FGFR) gene expression and splicing in ARPE-19 cells. This in vitro model partially mimics in vivo patterns, offering insights into FGFR regulation during differentiation.
Area of Science:
- Molecular Biology
- Cell Biology
- Developmental Biology
Background:
- Fibroblast Growth Factor Receptors (FGFRs) play crucial roles in development and tissue function.
- The ARPE-19 cell line, a model for retinal pigment epithelium, exhibits differentiation capabilities in vitro.
- Understanding FGFR regulation is key to comprehending cellular differentiation processes.
Purpose of the Study:
- To investigate the regulation of FGFR gene expression and FGFR1 mRNA alternative splicing during ARPE-19 cell differentiation in vitro.
- To compare in vitro differentiation patterns with in vivo human retinal pigment epithelium (RPE) FGFR expression.
Main Methods:
- ARPE-19 cells were cultured at varying densities for up to 14 months to induce differentiation.
- Quantitative PCR was used to measure FGFR expression and the ratio of FGFR1beta to FGFR1alpha splice variants.
- In vivo human RPE samples served as controls for comparison.
Main Results:
- Undifferentiated ARPE-19 cells primarily expressed FGFR1; differentiation led to FGFR2 expression.
- In vivo RPE samples showed FGFR1, FGFR2, and FGFR3 expression, with FGFR3 being minimal in vitro.
- The ratio of FGFR1beta to FGFR1alpha splice variants decreased with in vitro differentiation, approaching in vivo levels.
- FGF2 stimulation did not alter the FGFR1 splice variant ratio.
Conclusions:
- In vitro differentiation of ARPE-19 cells partially recapitulates in vivo FGFR expression and splicing patterns.
- The ARPE-19 cell line serves as a valuable model for studying the regulation of FGFR gene expression and splicing during cellular differentiation.
- Further studies can utilize this system to explore the mechanisms underlying FGFR regulation in differentiation.