| Project/Area Number |
17591825
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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 | Kanazawa University |
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Principal Investigator |
TOMOMI Higashide Ophthalmology, Lecturer, 医学部附属病院, 講師 (20291370)
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| Co-Investigator(Kenkyū-buntansha) |
OHKUBO Shinji Ophthalmology, Assistant Professor, 医学部附属病院, 助手 (90362003)
SUGIYAMA Kazuhisa Ophthalmology, Professor, 医学系研究科, 教授 (80179168)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2006: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2005: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | retinal ganglion cell / in vivo imaging / scanning laser ophthalmoscope / rat |
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| Research Abstract |
1) In vivo imaging and quantitative evaluation of rat retinal nerve fiber layer (RNFL) by scanning laser ophthalmoscopy (SLO)
Methods: Fundus images of both eyes were recorded over time by SLO using an argon blue laser (488 nm) in unilateral optic nerve crush or ischemia-reperfusion model. The focused plane was sequentially moved by changing the refractive values in the SLO setting. The range of refractive values (AF) in which RNFL reflex was clearly observed was determined. The RNFL thickness in retinal sections was measured and compared to the DF value. Results: The QF value was unchanged 1 week after the crush, but then decreased significantly after the second week, while it decreased significantly from the 1st week after the ischemia-reperfusion. The OF value correlated significantly with the histologically determined RNFL thickness.
2) In vivo imaging and counting of rat retinal ganglion cells (RGCs) by SLO
Methods: RGCs of Brown Norway rats were retrogradely labeled bilaterally with
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the fluorescent dye, 4-(4-(dihexadecylamino)styryl)-N-methylpyridinium iodine (DiA). RGCs were imaged in vivo with a SLO using an argon blue laser and optical filter sets for fluorescein angiography, before and 1, 2, and 4 weeks after the crush. Fluorescent cells were also counted on retinal flatmounts. An image overlay analysis was performed to check cell positions in the SLO images over time. Lectin histochemical analysis was performed to determine the relationship of microglia to the newly emerged DiA fluorescence detected by image overlay analysis after the optic nerve crush. Results: Fluorescent RGCs were visible in vivo by SLO. RGC survival decreased gradually after the crush. Newly emerged DiA fluorescence detected by image overlay analysis corresponded to fluorescent cells morphologically different from RGCs in the retinal flatmount and was colocalized mostly with lectin-stained microglial processes. RGC counts by SLO were comparable to those by retinal flatmounts.
3) Conclusions: The SLO is useful for in vivo imaging and quantitative evaluation of rat RNFL and RGCs, and therefore may be a valuable tool for monitoring RNFL and RGC changes over time in various rat models of RGC damage. Less
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