Paracrine Effects of Phosphorylated and Excreted FGF1 by Retinal Pigmented Epithelial Cells

Abstract

We have recently shown that both inhibition of endogenous Fibroblast growth factor (FGF) synthesis in non dividing lens epithelial cells (Renaud el al. J. Biol. Chem 1996, 271 : 2801–2811) and inhibition of secreted FGF1 in confluent quiescent retinal pigmented epithelial (RPE) cells (Guillonneau et al., Exp. Cell. Res. 1997, in press) induce rapid cell apoptosis. In addition, FGF2-stimulated release of endogenous FGF1 is associated with reduced apoptosis in RPE cells. We now show that a single addition of exogenous FGF2 to RPE cells induces after 4 days of culture, a great accumulation of FGF1 within the cells. Concomitantly we observe that FGF1 was released into the extracellular medium. Secreted FGF1 from RPE cells, purified from culture medium and added to either Go-arrested RPE or RMG cells at low plating density induced cell proliferation, whereas when it is added once to serum-depleted confluent RPE and RMG cells it prevented apoptosis. Both endogenous and secreted FGF1 are phosphorylated. In addition, FGF2 stimulated the production and release of phosphorylated FGF1 by RPE cells. We show that this secreted form of phosphorylated FGF1 binds to the high affinity tyrosine kinase receptors of RPE and RMG cells on retinal sections and to heparan sulfate proteoglycan in RPE cell extracellular matrix. In contrast to non-phosphorylated FGF1, phosphorylated secreted FGF1 was not degraded after internalization but accumulated within RPE and RMG cells, and is rapidly translocated to the nucleus suggesting a role in signal transduction and gene expression pathways. These results show that exogenous FGF2 activities might be mediated indirectly by phosphorylation and that secretion of FGF1 may function as a paracrine trophic factor for retinal cells.

Key Words:

• Fibroblast growth factor 1
• phosphorylation
• retinal pigmented epithelial cells