Genetic characterization of the multipotent dedifferentiated state of pigmented epithelial cells in vitro.
Retinal pigmented epithelial cells (PECs) of chicken embryos extensively and almost synchronously transdifferentiate into lens cells in medium containing phenylthiourea and testicular hyaluronidase, passing through the bipotent dedifferentiated state. We have isolated genes that are expressed specifically by either pigment or lens cells and analyzed their expression in the transdifferentiation process. The expression of some proto-oncogenes was also studied. In the dedifferentiation process, expression of the c-myc gene was enhanced and the transcription of PEC-specific genes (MMP115, pP344) was completely repressed. However, transcription of lens-specific genes (alpha-, beta- and delta-crystallins genes) remained silent in dedifferentiated pigment cells. Activation of len- or PEC-specific genes occurred only in conditions permissive for lens or PEC differentiation, respectively. These results indicated that lens transdifferentiation from PECs proceeds through a multipotent (or at least bipotent) intermediate cell state in which the c-myc gene is activated, but neither PEC- nor lens-specific genes are expressed.
Agata
K
K
Department of Developmental Biology, National Institute for Basic Biology, Okazaki, Japan.
Kobayashi
H
H
Itoh
Y
Y
Mochii
M
M
Sawada
K
K
Eguchi
G
G
D000818:Animals; D015152:Blotting, Northern; D002454:Cell Differentiation / Q000235:genetics; D002478:Cells, Cultured; D002642:Chick Embryo; D003459:Crystallins / Q000235:genetics; D005136:Eye Proteins / Q000235:genetics*; D015870:Gene Expression / Q000502:physiology; D015151:Immunoblotting; D010857:Pigment Epithelium of Eye / Q000166:cytology / Q000196:embryology* / Q000502:physiology; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics; D011519:Proto-Oncogenes / Q000235:genetics*
D003459:Crystallins; D005136:Eye Proteins; D016271:Proto-Oncogene Proteins c-myc