| Project/Area Number |
23KJ1280
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| Research Category |
Grant-in-Aid for JSPS Fellows
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| Allocation Type | Multi-year Fund |
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| Section | 国内 |
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| Review Section |
Basic Section 46010:Neuroscience-general-related
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| Research Institution | Kyoto University |
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Principal Investigator |
ZHOU CHUYING 京都大学, 高等研究院, 特別研究員(DC2)
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| Project Period (FY) |
2023-04-25 – 2025-03-31
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| Project Status |
Granted (Fiscal Year 2023)
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| Budget Amount *help |
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 2024: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2023: ¥900,000 (Direct Cost: ¥900,000)
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| Keywords | Nesprin-2 / Dynein / Kinesin-1 / Nucleus / Neuronal migration |
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| Outline of Research at the Start |
During mammalian brain development, neurons undergo active migration, accompanied with forward movements of the nuclei. This study aims to reveal how a nuclear membrane protein, Nesprin-2 generates forward neuronal nuclear movements by coordinated the activities of opposing microtubule motors, dynein and kinesin-1. By advanced live-cell imaging techniques and a thorough analysis of Nesprin-2-mediated intracellular cargo dynamics, a mechanistic understanding on the neuronal nuclear dynamics will be achieved, offering new insights into neurodevelopmental diseases.
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| Outline of Annual Research Achievements |
My major achievements are as followed: (1) biochemical experiments were performed to further analyze how Nesprin-2 binds to dynein complexes by utilizing a BicD2 stable knockout cell line, (2) protein structure prediction was used to understand the 3D conformation of Nesprin-2 protein, which shows an atypical arrangement of dynein-dynactin-binding motifs, (3) rescue experiments were performed using Nesprin-2 knockout cells and expected results were obtained, showing that binding to both microtubule motors is sufficient for the recovery of nuclear movements (4) movements of microtubules in migrating neurons were analyzed by photo-conversion experiments. Taken together, our results provide further insights into the role of Nesprin-2 as a coordinating adaptor for smooth nuclear transport.
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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
Although some modifications of experiment plan were made, the project overall is progressing smoothly.
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| Strategy for Future Research Activity |
During the current fiscal year, I found that Nesprin-2 drives active bidirectional movements over a prolonged period and keeps the cargo at a mobile state, while the forward movements of peri-nuclear microtubule tracks facilitate directional transport of the cargo during neuronal migration. I am now preparing manuscript for paper submission and the future experiment plans are as followed: (1) further characterization of Nesprin-2-dynein-dynactin-BicD2 binding through generating mutant molecules for biochemistry experiments, (2) intracellular cargo trafficking experiments with increased cargo size, (3) further characterization of microtubule sliding in migrating neurons.
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