Nano-imaging of the growth cone producing the driving force for nerve growth

2021 Fiscal Year Final Research Report

• Project/Area Number: 17KK0144
• Research Category: Fund for the Promotion of Joint International Research (Fostering Joint International Research)
• Allocation Type: Multi-year Fund
• Research Field: Neurochemistry/Neuropharmacology
• Research Institution: Niigata University
• Principal Investigator: Nozumi Motohiro 新潟大学, 医歯学系, 講師 (00420323)
• Project Period (FY): 2018 – 2021
• Keywords: 成長円錐 / 超解像陰影法 / 超解像顕微鏡 / オルガネラ / アクチン

Outline of Final Research Achievements

To observe the interaction between various organelles and the cytoskeleton in the growth cones that lead neurites, we applied super-resolution shadow imaging, SUSHI, to visualize the intracellular structure. Using cultured cells that cytosolically expressed the fluorescent protein Citrine, we could readily discriminate negative images of various organelles, such as mitochondria, endoplasmic reticulum, endosome, lysosome and also cytoskeleton. The live shadow imaging showed that vesicles arising following actin polymerization and organelles that repeatedly move to the leading edge against the actin retrograde flow. Mitochondrial fission was also detected immediately after a vesicle approached the mitochondrion in the growth cone. The super-resolution imaging is one of the best methods to visualize multiple organelles and cytoskeletons simultaneously.

Free Research Field

神経科学

Academic Significance and Societal Importance of the Research Achievements

超解像陰影法による細胞内の可視化は、関心のある細胞内構造を蛍光標識する従来の方法に比べて、簡便に多くのオルガネラや細胞骨格を同時検出できるだけでなく、非選択的な可視化によって思い込みに囚われない細胞内イメージングを実現できる。原法のSUSHI (super-resolution shadow imaging)と組み合わせることで、生きた組織内の細胞内外を同時にライブイメージングすることが可能になった。