2022 Fiscal Year Final Research Report
Development of a novel method for live cell imaging of intracellular and extracellular structures in brain tissue
| Project/Area Number |
21K19305
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| Research Category |
Grant-in-Aid for Challenging Research (Exploratory)
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| Allocation Type | Multi-year Fund |
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| Review Section |
Medium-sized Section 46:Neuroscience and related fields
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| Research Institution | Niigata University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
本多 敦子 新潟大学, 医歯学系, 特任助教 (40467072)
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| Project Period (FY) |
2021-07-09 – 2023-03-31
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| Keywords | 成長円錐 / 超解像顕微鏡 / SUSHI / COSHI / アクチン / ミトコンドリア |
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| Outline of Final Research Achievements |
Super-resolution shadow imaging (SUSHI) enables the acquisition of shadow images of brain tissue, including synapses, through fluorescent labeling of the extracellular space. In this study, we applied the principles of SUSHI to visualize intracellular structures, aiming to simultaneously image both intracellular and extracellular structures in living brain tissue. By employing this approach, the shadow imaging of growth cones, which are axonal tips, revealed a novel relationship between mitochondrial morphology and the actin cytoskeleton during nerve growth processes. To achieve this, a red fluorescent protein was introduced into fetal mouse brains through in utero electroporation. Through this method, we successfully captured intracellular structures of the neuron in the shadow images generated from red fluorescence signals. We also observed a growth cone extending its filopodium towards a neighboring cell, followed by cell-cell contact using our imaging technique.
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| Free Research Field |
神経科学
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| Academic Significance and Societal Importance of the Research Achievements |
本研究により、２種類の蛍光画像から、脳組織において軸索伸長や遊走で形態変化する神経細胞とその周りを取り囲む多様な細胞群との相互作用を捉えつつ、同時にオルガネラなどの細胞内構造の変化を可視化することが可能になった。今回試みた細胞内構造の可視化は神経細胞に限らず、線維芽細胞やグリオーマ株細胞にも適応できたことから、神経細胞からグリア細胞まで幅広い細胞種での細胞内外ライブイメージングに応用できると思われる。このライブイメージング法は脳の形成過程だけでなく、病態脳における様々な細胞集団の細胞内外動態を網羅的に可視化するツールとしての応用が期待される。
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