| Project/Area Number |
07457417
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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 | Juntendo University School of Medicine |
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Principal Investigator |
KANAI Atsushi Juntendo Univ., Ophthalmol., Prof., 医学部, 教授 (00053059)
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| Co-Investigator(Kenkyū-buntansha) |
FUJIKI Keiko Juntendo Univ., Opthalmol., Assistant Prof., 医学部, 講師
HOTTA Yoshihiro Juntendo Univ., Opthalmol., Assistant Prof., 医学部, 講師 (90173608)
YOKOYAMA Toshiyuki Juntendo Univ., Opthalmol., Assistant Prof., 医学部, 講師 (00191528)
NAKAYASU Kiyoo Juntendo Univ., Ophthalmol., Associate Prof., 医学部, 助教授 (10124976)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1996: ¥600,000 (Direct Cost: ¥600,000)
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| Keywords | cornea / cDNA / NADH ubiquinone / FKBP / thrombospondin / alpha1 type VIII collagen / corneal dystrophy / 角膜ジストロフィ / NADHユビキノ酸化還元酵素 / K-グリピカン / SPARC / 候補遺伝子 / 連鎖分析 / 遺伝子導入 / ウイルスベクター / 膠様滴状角膜ジストロフィ |
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| Research Abstract |
To obtain the candidate genes for the corneal dystrophies, plus/minus screening of the rabbit corneal cDNA library from the Department of Molecular Biology and Biochemistry, Okayama University Medical School was performed using corneal and iris RNA as probes. Thirteen clones corresponding to three ferritin H-chain, a NADH-ubiquinone oxidoreductase B22 subunit, an alpha 1 type VIII collagen, a 25KDa FKBP-506 binding protein (FKBP25), a thrombospondin 2 and six unknown clones were isolated. RT-PCR analysis showed that FKBP 12 and 25 highly express in the rabbit cornea, retina cerebrum, cerebellum. They also highly express in the cultured corneal epithelial cells. Although proteins tranlocated from these isolated mRNA are not corneal specific, they will play an important role in the cornea. None of the isolated known mRNAs does not map to the chromosome 5,16 or 20. We plan the further study including the chromosomal localization of the unknown clones using the BAC screening and FISH.We also randomly extracted the about 700 clones from the library, sequenced partially and done the homology search using the Internet service.
Since the confirmation of the diagnosis of the corneal dystrophies was required, we evaluated the 159 samples of the corneal button pathologically. We elaborately diagnosed the corneal dystrophies and found the Avellino corneal dystrophy which is the first report in Japan. We extracted the DNA from these patients and stocked them for the further analysis. We also completed the preparation for the basic study for the gene transfer to the corneal tissue.
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