| Project/Area Number |
17500199
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Neuroscience in general
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| Research Institution | University of Tsukuba |
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Principal Investigator |
CHIBA Chikafumi University of Tsukuba, Graduate School of Life and Environmental Sciences, Associate Professor (80272152)
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| Project Period (FY) |
2005 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,740,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥240,000)
Fiscal Year 2007: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2006: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2005: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | newt / retinal regeneration / retinal pigment epithelium cell / transdifferentiation / RPE65 / Notch signal / FGF2 / MEK / 網膜 / 再生 / 色素上皮細胞 / Musashi / RPE / Pax6 / Musashi1 / Notch-1 / Pax-6 / Musashi-1 |
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| Research Abstract |
In this study, as a ground-to elucidate the cellular and molecular mechanisms of retinal regeneration in the adult newt, we analyzed (1) the transdifferentiation process of the retinal pigment epithelium (RPE) cells using cellular and molecular markers and (2) the underlying molecular pathway by manipulating molecular functions.
Results of (1): we outlined the process of retinal regeneration as nine morphological stages and found that the transdifferentiation of RPE cells have no obvious dedifferentiation state.
Results of (2) Functional analyses of Pax6, Chx10-1, Notch signaling molecules, and neurogenin: we demonstrated that (I) Notch signaling is involved in neuronal differentiation during retinal regeneration and (ii) the adult newt RPE cells express both positive and negative regulators of neurogenesis, neurogenin 1 and Hes-1 genes. (iii) We tried microinjection of expression vectors into RPE cells in the eye of living animal, but we still continue to find better conditions because injection itself frequently resulted in cell death. (iv) We found that defect of Musashi 1 causes retinal degeneration.
Functional analyses of FGF2 using serum-free culture systems of RPE cells: we found that (I) FGF2 promotes dedifferentiation of RPE cells, proliferation and expression of pan-retinal neuron markers in vitro, (ii) transdifferentiation of RPE cells may be induced through an MEK pathway, (iii) this pathway is unlikely to be the same as embryonic/larval one (FGF2/FGFR-2/MEK/Pax6 pathway), and (iv) the embryonic/larval pathway may be reestablished within 10 days of induction.
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