Previous studies showed that lens epithelial cells proliferate rapidly in the embryo and that a lens mitogen, most likely derived from the blood, is present in the anterior chamber of the embryonic eye (Hyatt, G. A., and Beebe, D. C., Development 117, 701-709, 1993). Messenger RNAs for several growth factor receptors have been identified in embryonic lens epithelial cells. We tested several growth factors that are ligands for these receptors for their ability to maintain lens cell proliferation. Embryo serum, PDGF, GM-CSF, and G-CSF maintained lens cell proliferation, but NGF, VEGF, and HGF did not. This and a previous study (Potts, J. D., Harocopos, G. J., and Beebe, D. C., Curr. Eye Res. 12, 759-763, 1993) detected members of the Janus kinase family (Jaks) in the developing lens. Because Jaks are central players in the Jak-STAT-signaling pathway, we identified STAT proteins in the lens and tested whether they were phosphorylated in response to mitogens. STAT1 and STAT3, but not STAT 5 were detected in chicken embryo lens epithelial cells. Only STAT3 was found in terminally differentiated lens fiber cells. STAT1 and STAT3 were phosphorylated in lens cells analyzed immediately after removal from the embryo and when lens epithelial explants were treated with embryo serum, PDGF, or GM-CSF, but not with NGF. Chicken embryo vitreous humor or IGF-1, factors that stimulate lens cell differentiation, but not proliferation, did not cause STAT phosphorylation. When lens epithelial cells were cultured for 4 h in unsupplemented medium, STAT1 and STAT3 declined to nearly undetectable levels. Treatment with PDGF or embryo serum for an additional 15 min restored STAT1 and -3 levels. This recovery was blocked by cycloheximide, but not actinomycin D, suggesting that STAT levels are regulated at the level of translation. STAT levels were maintained in epithelial explants by lens mitogens, but not by factors that stimulated lens fiber differentiation. Both factors that stimulated lens cell proliferation and those that caused fiber differentiation protected cultured lens epithelial cells from apoptosis. These data suggest that the factor(s) responsible for lens cell proliferation in vivo activates the Jak-STAT-signaling pathway. They also indicate that growth factors maintain STAT protein levels in lens epithelial cells by promoting the translation of STAT mRNA, an aspect of STAT regulation that has not been described previously. Signaling by most of the growth factors and cytokines known to activate the Jak-STAT pathway has been disrupted in mice by mutation or targeted deletion. Consideration of the phenotypes of these mice suggests that the factor responsible for lens cell proliferation in vivo may be a growth factor or cytokine that has not yet been described.
Copyright 1998 Academic Press.