2007 Fiscal Year Final Research Report Summary
Generation and characteristics of transgenic rabbit model of retinal degeneration
| Project/Area Number |
18591913
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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 | Nagoya University |
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Principal Investigator |
KONDO Mineo Nagoya University, Graduate School of Medicine, Associate Professor (80303642)
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| Co-Investigator(Kenkyū-buntansha) |
TERASAKI Hiroko Nagoya University, Graduate School of Medicine, Professor (40207478)
NAKAMURA Makoto Nagoya University, Graduate School of Medicine, Associate Professor (60283438)
ITO Yasuki Nagoya University, Graduate School of Medicine, Designeted Associate Professor (10313991)
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| Project Period (FY) |
2006 – 2007
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| Keywords | Transgenic / Rabbit / Animal model / Retinal degeneration / Photoreceptor / Retinitis piementosa / Electroretinoeram / Medium-sized animal |
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| Research Abstract |
Purpose of the present research was to generate a rabbit that expresses a mutant rhodopsin gene, and to characterize the progressive retinal degeneration using electroretinography(ERG), histology, and immunohistochemistry. The rabbit DNA clone that contains a rhodopsin gene was identified in a NZW rabbit BAC library. The Pro347Leu mutation of the rhodopsin gene was introduced homologous recombination in E. col. and these BACs were microinjected into fertilized eggs. Transgene-positive animals were identified by Southern blot analysis. The expression level of transgene in the retinal tissue was measured by qualitative real-time RT-PCR. Scotopic and photopic ERGs elicited by different stimulus intensities were recorded. Paraffin retinal sections were stained with hematoxylin and eosin. Rhodspin and lectin immunohistochemistory, and electronmicroscopy was also performed. Ten transgene-positive founders were identified, six of which passed transgenes to offsprings. The expression level of the transgene ranged from 7-80% for six transgenic lines, and was roughly correlated with transgene copy number and speed of retinal degeneration. In line 7, the rod ERG a-wave was nearly extinguished by 12-month-old, but the cone a-wave retained 60% of controls. Retinal histology demonstrated a progressive loss of photoreceptors and a shortening of the outer segments, and ONL at visual streak showed only 1-2 laws by 12 weeks. This is the first transgenic rabbit model of retinal degeneration. The speed of retinal degeneration was dependant on the expression level of the transgene. Because the rabbits have large eyes and are easy to handle, these animals will be a useful model to stud the pathophysiology and treatment of retinal degeneration
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