Basic fibroblast growth factor as one of the essential factors regulating lens transdifferentiation of pigmented epithelial cells.
In vitro transdifferentiation of retinal pigmented epithelial cells of the chick embryo into lens cells can be markedly enhanced by culture in the presence of testicular hyaluronidase and phenylthiourea. Since the commercial preparations of hyaluronidase that had previously been used were very crude, a search for the actual effective molecule(s) enhancing lens transdifferentiation was conducted. First, we purified the enzyme and tested the effect of the purified hyaluronidase. Highly purified hyaluronidase itself did not enhance lens transdifferentiation. The crude hyaluronidase was then separated according to affinity with heparin, considering the possibility that the fibroblast growth factor (FGF) is contained in the crude hyaluronidase. Transdifferentiation-enhancing activity was detected in the fraction which was bound to heparin and eluted with 2 M NaCl, where no hyaluronate-degrading activity existed. Analysis of the fraction by SDS-PAGE revealed the existence of an 18 kDa protein whose NH2-terminal sequence was identical to that of basic FGF. The basic FGF derived from bovine brain also enhanced lens transdifferentiation of pigmented epithelial cells. These findings suggest that basic FGF must play a major role in enhancing transdifferentiation of pigmented epithelial cells to lens cells.
Hyuga
M
M
Department of Molecular Biomechanics, School of Life Science, Graduate University for Advanced Studies, Okazaki, Japan.
Kodama
R
R
Eguchi
G
G
D000595:Amino Acid Sequence; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002642:Chick Embryo; D002846:Chromatography, Affinity; D003459:Crystallins / Q000032:analysis / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D006821:Hyaluronoglucosaminidase / Q000302:isolation & purification / Q000494:pharmacology; D007908:Lens, Crystalline / Q000166:cytology / Q000196:embryology / Q000502:physiology*; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010670:Phenylthiourea / Q000494:pharmacology; D010857:Pigment Epithelium of Eye / Q000166:cytology* / Q000187:drug effects; D012756:Sheep; D013737:Testis / Q000201:enzymology
D003459:Crystallins; D016222:Fibroblast Growth Factor 2; D010670:Phenylthiourea; D006821:Hyaluronoglucosaminidase