2019 Fiscal Year Annual Research Report
神経ネットワーク形成における微小管結合タンパク質タウのアイソフォーム特異的な役割
| Project/Area Number |
19J13396
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| Research Institution | Tokyo Metropolitan University |
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Principal Investigator |
DILINA Tuerde 首都大学東京, 大学院理工学研究科, 特別研究員(DC2)
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| Project Period (FY) |
2019-04-25 – 2021-03-31
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| Keywords | tau / ASOs / isoform / development / axon / spine formation / synapse / dendrite arborization |
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| Outline of Annual Research Achievements |
In this year, I studied the role of 3-repeat (3R) and 4-repeat (4R) isoforms of tau in neuronal development in vitro and in vivo. Firstly, I have confirmed the most suitable concentration of antisense oligonucleotides (ASOs) for 3R- or 4R-tau in vitro and in vivo experiments. Next, to reveal isoform-specific function of tau in neurons during development, I analyzed the morphology, including the length of axons, dendritic arborization and synapse formation, of neurons expressing 3R- or 4R-tau. Interestingly, while the length of neurites and dendrite’s arborization were not different between 3R- or 4R-ASOs transfected neurons, the number of synapses and spine formation were significant different between them. In 3R-ASOs transfected neurons, spine maturation was delayed but the number of spine and synapses were not affected. In contrast, the maturation of dendritic spine was not affected by 4R-ASOs transfection but neurons lost synapses significantly. In vivo experiment, brain tissues of those administered mice were cleared with CLARITY and prepared for observation of neuronal morphology by the 3D imaging technique.
During this year, I found that tau isoforms differently affect synapse formation during neuronal development in vitro, although the length of neurites and dendritic arborization were not affected.
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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
The experimental results and progress are consistent with the plan.
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| Strategy for Future Research Activity |
I would like to examine the role of isoform alternation of tau in regulating postnatal neuronal function in vivo. AAV expressing TET-inducible GCaMP6 will be injected into the cerebral cortices of htau mice. The neuronal activity will be measured with in vivo calcium imaging using a two-photon microscopy. The two-photon calcium imaging of individual dendrites and axons will provide us the functional difference of neurons which treated with 3R-, 4R-, or control-ASO.
Write a manuscript and submit it to journal for publication.
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