Elucidation of the molecular basis of the ultra-early stage of endocytosis by high-resolution observation of cell membrane dynamics
Project/Area Number |
19K22506
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Research Category |
Grant-in-Aid for Challenging Research (Exploratory)
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Allocation Type | Multi-year Fund |
Review Section |
Medium-sized Section 48:Biomedical structure and function and related fields
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Research Institution | Hokkaido University |
Principal Investigator |
Ohba Yusuke 北海道大学, 医学研究院, 教授 (30333503)
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Project Period (FY) |
2019-06-28 – 2021-03-31
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Project Status |
Completed (Fiscal Year 2020)
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Budget Amount *help |
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2020: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2019: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
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Keywords | 細胞膜 / 高速分子間力顕微鏡 / 薄層遮光照明法 / 相関イメージング / correlative imaging / 高速原子間力顕微鏡 / 薄層 斜光照明法 |
Outline of Research at the Start |
細胞膜の形態と成分のダイナミックな変化は、細胞が周囲環境変化に応答した物質の取り込みやシグナル伝達を発動するために重要である。本研究では、(1) 高速原子間力顕微鏡による細胞膜微細構造の物理的高分解能計測と、(2) 薄層斜光照明法と蛍光異方性を用いた細胞膜形態の回折限界を超えた光学的計測 とを同時に行う。この世界でオンリーワンの高分解能計測法を用いたバイオイメージングにより、エンドサイトーシスを効率的に行うために予めプログラムされたドメインを解析し、細胞膜形態変化の分子基盤の解明および「細胞膜動態学の創出」を目指す。
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Outline of Final Research Achievements |
In this study, we utilized correlative imaging by hybrid microscopy combined with live-cell high-speed atomic force microscopy and fluorescence microscopy to observe and analyze the dynamics of cell membranes and molecules associated with clathrin-dependent endocytosis. We found that clathrin-coated pit formation induced by growth factor stimulation appears in a certain localized region of the plasma membrane and the morphology of the pit is not limited to circular (spherical), but can also be elliptical and divisive. Wey also found that Src is involved in such phenomena.
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Academic Significance and Societal Importance of the Research Achievements |
細胞内と細胞外環境を隔てる脂質二重膜であると同時に、細胞内外のコミュニケーションを司るインターフェースである細胞膜の3次元的なダイナミクスに関する知見はこれまで限られていた。本研究では、これまでの動態観察の分解能を10倍向上した高速AFMを用いることで、細胞と組織の活動や運命の決定に重要な細胞外からの刺激に応じた細胞膜の形態と成分の動的変化の解析を可能とし、かつその分子メカニズムの一端を明らかにした。
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Report
(3 results)
Research Products
(22 results)
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[Journal Article] Calcium Wave Promotes Cell Extrusion2020
Author(s)
Takeuchi Y、Narumi R、Akiyama R、Vitiello E、Shirai T、Tanimura N、Kuromiya K、Ishikawa S、Kajita M、Tada M、Haraoka Y、Akieda Y、 Ishitani T、Fujioka Y、Ohba Y、Yamada S、Hosokawa Y、Toyama Y、Matsui T、Fujita Y
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Journal Title
Current Biology
Volume: 30
Issue: 4
Pages: 670-681.e6
DOI
NAID
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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[Journal Article] Localization of BCR-ABL to Stress Granules Contributes to Its Oncogenic Function2019
Author(s)
Sayaka Kashiwagi, Yoichiro Fujioka, Takeshi Kondo, Aya O. Satoh, Aiko Yoshida, Mari Fujioka, Hitoshi Sasajima, Maho Amano, Takanori Teshima, Yusuke Ohba
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Journal Title
Cell Structure and Function
Volume: 44
Issue: 2
Pages: 195-204
DOI
NAID
ISSN
0386-7196, 1347-3700
Related Report
Peer Reviewed / Open Access
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[Journal Article] Folding Latency of Fluorescent Proteins Affects the Mitochondrial Localization of Fusion Proteins2019
Author(s)
Sayaka Kashiwagi, Yoichiro Fujioka, Aya O. Satoh, Aiko Yoshida, Mari Fujioka, Prabha Nepal, Atsushi Tsuzuki, Ozora Aoki, Sarad Paudel, Hitoshi Sasajima, Yusuke Ohba
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Journal Title
Cell Structure and Function
Volume: 44
Issue: 2
Pages: 183-194
DOI
NAID
ISSN
0386-7196, 1347-3700
Related Report
Peer Reviewed / Open Access
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[Journal Article] Development of immortalized human tumor endothelial cells from renal cancer2019
Author(s)
Nako Maishi, Hiroshi Kikuchi, Masumi Sato, Hiroko Nagao-Kitamoto, Dorcas A. Annan, Shogo Baba, Takayuki Hojo, Misa Yanagiya, Yusuke Ohba, Genichiro Ishii, Kenkichi Masutomi, Nobuo Shinohara, Yasuhiro Hida, Kyoko Hida
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Journal Title
Int J Mol Sci
Volume: 20
Issue: 18
Pages: 4595-4595
DOI
Related Report
Peer Reviewed / Open Access
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[Journal Article] Clinical efficacy and safety of first-line nilotinib therapy and evaluation of the clinical utility of the FRET-based drug sensitivity test.2019
Author(s)
Kondo T, Fujioka M, Fujisawa S, Sato K, Tsuda M, Miyagishima T, Mori A, Iwasaki H, Kakinoki Y, Yamamoto S, Haseyama Y, Ando S, Shindo M, Ota S, Nishio M, Kurosawa M, Ohba Y, Teshima T, on behalf of the North Japan Hematology Study Group (NJHSG).
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Journal Title
International Journal of Hematology
Volume: 110
Issue: 4
Pages: 482-489
DOI
Related Report
Peer Reviewed / Open Access
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