In vivo imaging of retinal ganglion cell function using adeno-associated virus

• Project/Area Number: 16K15730
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Ophthalmology
• Research Institution: Tohoku University
• Principal Investigator: Nakazawa Toru 東北大学, 医学系研究科, 教授 (30361075)
• Co-Investigator(Kenkyū-buntansha): 西口 康二 東北大学, 医学系研究科, 准教授 (30447825) ; 藤田 幸輔 東北大学, 医学系研究科, 助手 (80708115) ; 佐藤 孝太 東北大学, 医学系研究科, 助教 (50732327) ; 横山 悠 東北大学, 大学病院, 助教 (00597312) ; 村山 奈美枝 東北大学, 大学病院, 研究員 (60597516)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000); Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: 網膜神経節細胞 / 緑内障 / アデノ随伴ウイルス / E-SARE / RGC / in vivoイメージング

Outline of Final Research Achievements

Glaucoma, a leading cause of blindness worldwide, is characterized by progressive loss of the retinal ganglion cells (RGCs). To study neuronal activity in pathogenesis of RGC death, we developed in vivo imaging for quantitating activities of neuronal activity. E-SARE drives neuronal activity dependent gene expression at high levels.
The E-SARE-driven fluorescent protein reporters was packaged into AAV. The temporal activity was monitored using confocal ophthalmoscopy. Dark-adapted animals were kept in dark for 48 hours before use. Light-adapted animals were kept under a constant light for 6 hours after dark-adaptation for 48 hours before use. E-SARE reporter expression was induced by light stimulation, and localized in RGC. Increased reporter activity occurred by 2 hours after light stimuli, and decreased at 2hours after dark-adaptation showing 44% decreases. This E-SARE reporter in vivo cellular imaging is a usuful tool to assess the RGCs function in retinal diseases.