Neural cell differentiation from retinal pigment epithelial cells of the newt: an organ culture model for the urodele retinal regeneration.
Transdifferentiation from retinal pigment epithelium (RPE) to neural retina (NR) was studied under a new culture system as an experimental model for newt retinal regeneration. Adult newt RPEs were organ cultured with surrounding connective tissues, such as the choroid and sclera, on a filter membrane. Around day 7 in vitro, lightly pigmented "neuron-like cells" with neuritic processes were found migrating out from the explant onto the filter membrane. Their number gradually increased day by day. BrdU-labeling study showed that RPE cells initiated to proliferate under the culture condition on day 4 in vitro, temporally correlating to the time course of retinal regeneration in vivo. Histological observations of cultured explants showed that proliferating RPE cells did not form the stratified structure typically observed in the NR but they rather migrated out from the explants. Neuronal differentiation was examined by immunohistochemical detection of various neuron-specific proteins; HPC-1 (syntaxin), GABA, serotonin, rhodopsin, and acetylated tubulin. Immunoreactive cells for these proteins always possessed fine and long neurite-like processes. Numerous lightly pigmented cells with neuron-like morphology showed HPC-1 immunoreactivity. Fibroblast growth factor-2 (FGF-2), known as a potent factor for the transdifferentiation of ocular tissues in various vertebrates, substantially increased the numbers of both neuron-like cells and HPC-1-like immunoreactive cells in a dose-dependent manner. These results indicate that our culture method ensures neural differentiation of newt RPE cells in vitro and provides, for the first time, a suitable in vitro experimental model system for studying tissue-intrinsic factors responsible for newt retinal regeneration.
Ikegami
Yoko
Y
Developmental Neurobiology Laboratory, Department of Biological Sciences, Faculty of Science, Nara Women's University, Nara 630-8506, Japan.
Mitsuda
Sanae
S
Araki
Masasuke
M
D000818:Animals; D000963:Antimetabolites; D001973:Bromodeoxyuridine; D002454:Cell Differentiation; D002455:Cell Division / Q000502:physiology; D002478:Cells, Cultured; D003238:Connective Tissue / Q000502:physiology; D004847:Epithelial Cells / Q000166:cytology* / Q000502:physiology; D005260:Female; D007150:Immunohistochemistry; D008858:Microscopy, Phase-Contrast; D009416:Nerve Regeneration / Q000502:physiology*; D009924:Organ Culture Techniques; D010857:Pigment Epithelium of Eye / Q000166:cytology* / Q000502:physiology; D012455:Salamandridae / Q000502:physiology*
D000963:Antimetabolites; D001973:Bromodeoxyuridine