Exploration of regeneration of newt vision at the molecular level
| Project/Area Number |
15570062
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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 |
Animal physiology/Animal behavior
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| Research Institution | Osaka University |
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Principal Investigator |
HISATOMI Osamu Osaka University, Graduate School of Science, Associate Professor, 大学院・理学研究科, 助教授 (60231544)
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| Co-Investigator(Kenkyū-buntansha) |
TOKUNAGA Fumio Osaka University, Graduate School of Science, Professor, 大学院・理学研究科, 教授 (80025452)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2004: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2003: ¥2,600,000 (Direct Cost: ¥2,600,000)
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| Keywords | retina / EST / regeneration / newt / signal transduction / gene manupulation / expression |
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| Research Abstract |
In this project, we attempted to uncover the genes responsible for regeneration of newt retina. Our results can be summarized as follows.
DD750 gene identified by differential display (DD) method was introduced into E-coli cells and recombinant DD750 protein was expressed. Anti-DD750 antiserum recognized no RPE cells in normal retina but reacted with RPE cells several days after surgical removal of the retina. It is likely that DD750 play some roles in retinal regeneration.
A cDNA library (regenerating retinal cDNA library) were constructed using mRNAs isolated from regenerating retinas at 18-19 days after the surgical removal of the normal retina. Our sequence analysis of the regenerating cDNA library revealed that genes expressed in the regenerating retinal cells are apparently distinct from those in neither neural retina nor RPE cells. To understand the role of genes during transdifferentiation of RPE cells, we explored expression of several genes (otx, stathmin, thymosin and B-FABP etc.) by RT-PCR, immunohistochemistry and in situ hybridization. Our results indicate that spatiotemporal regulation of Otx2 expression is consistent with those of RPE markers. Otx2 may play a pivotal role in maintenance and specification of RPE cells during neural retina regeneration.
In order to knock down stathmin gene, we made a morpholino oligonucleotide (Mo-oligo) corresponding to an antisense sequence of stathmin gene, and injected it in the vitreous of newt eyes. The effect of the Mo-oligo on retinal structure was observed at 2 days after the injection. We settled several conditions that Mo-oligo was effectively introduced in retinal cells.
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Report
(3 results)
Research Products
(6 results)
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[Journal Article] Downregulation of Otx2 in the dedifferentiated RPE cells of regenerating newt retina.2005
Author(s)
Sakami, S., Hisatomi, O., Goto, T., Sakakibara, S.Liu, J.Reh, T.A., Tokunaga, F.
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Journal Title
Dev.Brain Res. 155(1)
Pages: 49-59
Description
「研究成果報告書概要(欧文)」より
Related Report
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