| Project/Area Number |
15591874
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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 |
Ophthalmology
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| Research Institution | Okinaka Memorial Institute Medical Research (2004)
Jichi Medical University (2003) |
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Principal Investigator |
MORI Mikiro Okinaka Memorial Institute for Medical Research, Researcher, 研究員 (00240721)
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| Co-Investigator(Kenkyū-buntansha) |
TSURU Tadahiko Jichi Medical School, Ophthalmology, Professor, 眼科, 教授 (90126128)
OBATA Hiroto Jichi Medical School, Ophthalmology, Assistant Professor, 眼科, 講師 (80224301)
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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 |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2004: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2003: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | Retinal pigment epitheium / Corneal epithelium / Knockout mouse / Cre / RXRα / Trasgenic mous / ノックアウト / loxP / トランスジェニック / タモキシフェン |
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| Research Abstract |
1.Knockout mouse system specific for the retinal pigment epithelium (RPE)
A transgenic mouse line was generated using a tyrosinase-related protein 1 (TRP1) promoter and a Cre recombinase gene and subsequently crossed with a line with floxed retinoid X receptor a (RXRα) gene to generate a TRP1-Cre : RXRα^<f1/f1> mouse line. In this line, the RXRα gene and protein were ablated in the RPE. The RPE was morphologically disorganized and functionally abnormal with decreases in visual retinoid cycle-related proteins, such as RPE65, CRALBP, and RGR. The neural retina showed shortening and degeneration of the outer segments with decreased responses in electroretinograms. These results indicated that an RPE-specific knockout mouse system is established and that RXRα is required in development and maintenance of the RPE in mice.
A transgenic mouse line was generated using the TRP1 promoter and a tamoxifen-dependent Cre (Cre-ERT2) and analyzed by crossing with a Z/AP reporter mouse line. Intraperitoneal administration of 1 mg tamoxifen caused Cre-mediated DNA recombination in about 70% of the RPE cells and less than 1% of the photoreceptor cells but not in other tissues of the body. The TRP1-CreERT2 mouse line was shown to be useful as a system for RPE specific inducible gene disruption in vivo (submitted).
2.Knockout mouse system specific for the corneal epithelium
Cre-ERT2 transgenic mouse line was generated using a promoter for keratin 14 (K14) and analyzed. Topical administration of tamoxifen to the eye surface resulted in gene disruption in less than 10% of the corneal epithelial cells and some scattered cells in the epithelia of skin and gastrointestinal epithelia. The results indicated that another promoter should be considered for increasing efficiency and specificity of corneal epithelium-specific gene disruption.
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