Journal
GENESIS
Volume 51, Issue 6, Pages 410-419Publisher
WILEY
DOI: 10.1002/dvg.22378
Keywords
retina regeneration; Xenopus laevis; Rax; Pax6; transgenic xenopus; tissue culture
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Funding
- Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan
- Nara Women's University
- Grants-in-Aid for Scientific Research [23570256] Funding Source: KAKEN
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The whole retina regenerates from retinal pigmented epithelial (RPE) cells by transdifferentiation in the adult newt and Xenopus laevis when it is surgically removed. We produced a transgenic animal line, in which EGFP expression is under the control of Rax pomotor. Using F1 and F2 generations, we analyzed Rax-EGFP expression during retinal regeneration in a tissue culture model. In the culture, 4 zones were distinguished as RPE cells migrating outwards from the periphery of the explant: the explant zone, epithelial zone, transition zone and differentiation zone. Expression of transcription factors such as Pax6 and Rax-EGFP was observed in different zones. Rax-EGFP expression preceded Pax6 expression, and the expression of both genes occurred in RPE cells that had lost contact with the basement membrane facing the choroid. We have developed a new culture method in which RPE tissues are embedded in Matrigel. This method has many advantages over the previous gel-overlay method to reproduce construction of 3D-retinal structures and clearly showed that RPE cells need to be detached from the choroid before entering the regeneration pathway. The present results indicate that the temporal changes in cell-cell and cell-extracellular matrix interactions regulate transdifferentiation. genesis 51:410-419. (c) 2013 Wiley Periodicals, Inc.
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