| Project/Area Number |
14370558
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Ophthalmology
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| Research Institution | Shimane University (2004)
Shimane Medical University (2002-2003) |
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Principal Investigator |
HASHIMOTO Ryuju Shimane University, Faculty of Medicine, Department of Anatomy, Instructor, 医学部, 助手 (90252907)
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| Co-Investigator(Kenkyū-buntansha) |
OTANI Hiroki Shimane University, Faculty of Medicine, Department of Anatomy, Professor, 医学部, 教授 (20160533)
HATTA Toshihisa Shimane University, Faculty of Medicine, Department of Anatomy, Associate Professor, 医学部, 助教授 (20238025)
UDAGAWA Jun Shimane University, Faculty of Medicine, Department of Anatomy, Instructor, 医学部, 助手 (10284027)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥11,700,000 (Direct Cost: ¥11,700,000)
Fiscal Year 2004: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2003: ¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 2002: ¥6,000,000 (Direct Cost: ¥6,000,000)
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| Keywords | mouse embryo / lens / FoxE3 / adenovirus / retrovirus / dyl / レポーター遺伝子 / β-galactosidase / Xgal染色 / βgalactosidase / Xqal染色 |
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| Research Abstract |
We aimed to induce ectopic lens which develops from the surface ectoderm by transferring the genes of transcription factors that are involved in eye development. To determine the optimal timing of transferring genes, we injected adenovirus which was combined with LacZ gene as a reporter gene to the amniotic fluid of mouse embryos from embryonic day (E) 8 to E13. We collected the infected embryos on E13 and E14. The infected cells were detected by X-gal staining and immunohistochemistry with anti-β galactosidase antibody. We could detect infected cells of the pigment layer and retina in the embryos injected on E8 and detected the infected mesenchyme between the lens and the retina excluding lens fibers and retina on E9. Some lens fiber cells in the embryos injected on E11 were infected. We injected the retrovirus combined with LacZ and collected mouse embryos at E14.5. Many lens fiber cells, some lens epithelial cells and some pigment epithelial cells of the retina in the embryos injected at E10.0 were infected by the retrovirus and total lens and a few pigment epithelia at E10.5 were infected. We decided the period of virus injection from E9.5 to E11.0. We next made adenovirus into which the construct composed of FoxE3, its regulatory region and SV40 poly A were integrated. This construct had been confirmed to rescue microphhalmia of dyl/dyl mice, the mutant of FoxE3. We made the adenovirus including this construct and concentrated supernatant of medium containing the adenovirus. We injected the virus media to the amniotic fluid of dyl/dyl embryos at E10.5 and collected the infected embryos at E15.5. The eyeball of the infected embryos did not show adhesion of the cornea and lens epithelia, but vacuoles and disturbed lens fibers were observed, suggesting the partial rescue of lens deformity.
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