| 研究課題/領域番号 |
20K06865
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| 研究種目 |
基盤研究(C)
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| 配分区分 | 基金 |
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| 応募区分 | 一般 |
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| 審査区分 |
小区分46010:神経科学一般関連
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| 研究機関 | 国立遺伝学研究所 |
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研究代表者 |
ZHU YAN 国立遺伝学研究所, 遺伝形質研究系, 助教 (50464235)
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| 研究期間 (年度) |
2020-04-01 – 2023-03-31
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| 研究課題ステータス |
交付 (2021年度)
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| 配分額 *注記 |
4,420千円 (直接経費: 3,400千円、間接経費: 1,020千円)
2022年度: 1,040千円 (直接経費: 800千円、間接経費: 240千円)
2021年度: 1,560千円 (直接経費: 1,200千円、間接経費: 360千円)
2020年度: 1,820千円 (直接経費: 1,400千円、間接経費: 420千円)
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| キーワード | metabolic pathway / neuronal migration / bioenergetics / ATP sensor / Bioenergetics / metabolic pathways / Neuronal migration / chain migration / energy status / energy sensor |
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| 研究開始時の研究の概要 |
All cellular activities consume energy which is supplied by metabolic pathways. Status of energy consumption both reflects and controls cell behaviors. This research investigates the dynamics of energy status and the metabolic pathways during the development of mammalian brains.
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| 研究実績の概要 |
This research has two objectives: (1) visualize and quantify the energy status of chain migrating neurons; (2) unravel the metabolic sources that fuel neuronal migration.
For objective (1), while I have established a system to express genetically encoded ATP sensors (ATeam) in vivo and in vitro, progress in FRET-based imaging of intracellular ATP is hindered by technical difficulties. For objective (2), I have generated a mouse knockout line with deficiency in a transcription factor known to promote OXPHOS metabolic pathway during development. Phenotypic analysis of the mutant line has shown defects in neuronal migration and/or their termination of migration.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
4: 遅れている
理由
The progress of this study is delayed.
The FRET-based ATP sensor imaging is delayed due to a broken down of a laser line of our common confocal imaging system and the time it took to replace the laser.
Another reason that delayed this project is due to COVID-related family issues.
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| 今後の研究の推進方策 |
(1)I will perform further phenotype analysis of the knockout mice generated to understand the defects in detail, including morphological changes, dendritic pattern and axon projection. As the knockout mice die at birth, I will also employ CRISPR/CAS9 mediated gene editing to knockout the gene of interest via local in utero electroporation and observe the effect in postnatal samples.
(2)I will perform manipulation of metabolic pathways using pharmacological reagents and examine the consequences of such manipulations on chain neuronal migration using in vitro whole tissue culture system.
(3)I will introduce the fluorescent ATP sensors into the migrating neurons and image ATP levels in both fixed and living conditions.
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