2020 Fiscal Year Research-status Report
iPS細胞由来網膜移植にて視機能再建のための変性網膜の視覚回路の解析と最適化
| Project/Area Number |
20K18403
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| Research Institution | Osaka University |
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Principal Investigator |
Tu HungYa 大阪大学, 蛋白質研究所, 助教 (10780835)
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| Project Period (FY) |
2020-04-01 – 2023-03-31
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| Keywords | retinal degeneration / horizontal cell / bipolar cell / ribbon synapse / organoid transplantation |
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| Outline of Annual Research Achievements |
To document the intrinsic property of horizontal cells (HCs) and to clarify their roles in ribbon synapse formation/preservation, the conditional HC labeling mice cross-bred into rd1 background with severe photoreceptor degeneration have been established. The conditional labeling protocol has then been optimized as the tamoxifen administration proves to be highly toxic with lethality for adult mice with or without photoreceptor degeneration.
The design of conditional HC labeling was also applied to induce the conditional HC deletion. The preliminary data by immunostaining shows that in normal retina the ribbon synapses and bipolar dendritic extension were rather preserved even without healthy HCs (or vice versa), while in the rd1 retinas with HC abolishment there were almost no residual synapse or potential dendritic contact detected.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
Interrupted by the pandemic situation, the progress of this project has been slightly delayed. However, the mouse lines and tamoxifen administration protocol have been established and stored with abundant numbers for stable and repetitive trials of experiments and transplantation in the near future.
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| Strategy for Future Research Activity |
The established horizontal cell conditional knockout mice will be used for mouse retinal organoid transplantation to clarify the role of HCs in synapse regeneration in comparison with their littermate without HC deletion as the ideal control.
On the other hand, HCs with specific labeling will be targeted for patch clamp recording to characterized their intrinsic property and synaptic inputs in normal and degenerated retinas.
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| Causes of Carryover |
The residual amount simply resulted from the unexpected pandemic situation and importation delay, which will be recovered in the next fiscal year as the experiment schedule has been modified to fit the current life situation.
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