2020 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 | Keio University |
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Principal Investigator |
DILINA Tuerde 慶應義塾大学, 医学部, 特別研究員(PD)
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| Project Period (FY) |
2019-04-25 – 2021-03-31
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| Keywords | neuron connection / GRAPHIC / ventral tegmental area / nucleus accumbens (NAc) / dopamine / tau isoform / synaptic interaction / non-synaptic interaction |
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| Outline of Annual Research Achievements |
To clarify the tau isoform specific function in neuronal connection in vivo, I tried to apply the GRAPHIC system to label all neuron to neuron connections which is new technique based on biomolecular fluorescence complementation (BiFC) and has been developed to label all neurons connections.
Firstly, to determine whether the GRAPHIC system could able to detect all neuronal connections that included both synaptic and non-synaptic, I applied the GRAPHIC system to the dopamine mesocorticolimbic pathway, the input from the ventral tegmental area (VTA) into nucleus accumbens (NAc), because of previous studies assumed that dopamine neurons could be formed both synaptic and non-synaptic communication with other neurons. Dopamine axons of VTA labeled by expressing C- GRAPHIC binds with AAV- EF1a-DIO-mCherry and co-expressing with AAV-TH-Cre, and neurons in NAc labeled by expressing N- GRAPHIC-mTagBFP2 with hSynapsinI promoter. After one month virus post-infection, I confirmed that the GRAPHIC signal expressed under mCherry (the source of dopamine neuron) and blue mTagBFP2 (the target neuron) colors as well as expressed with or without postsynaptic density, which indicated the GRAPHIC system could successfully label synaptic and non-synaptic connections sites.
During this year, I tried to re-design the GRAPHIC system for my experimental propose and tested its efficiency in vivo. By establishing the GRAPHIC system in the dopaminergic pathway, I showed the evidence for the first time the dopamine neurons make both synaptic and non-synaptic interactions with other neurons.
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| Research Progress Status |
令和2年度が最終年度であるため、記入しない。
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| Strategy for Future Research Activity |
令和2年度が最終年度であるため、記入しない。
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