Three-dimensional ultrastructural analysis of enteric glial cells with Cryo-fixation
| Project/Area Number |
17KK0191
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| Research Category |
Fund for the Promotion of Joint International Research (Fostering Joint International Research)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Gastroenterology
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| Research Institution | Nagoya University |
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Principal Investigator |
Tamada Hiromi 名古屋大学, 医学系研究科, 助教 (60712817)
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| Project Period (FY) |
2018 – 2020
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| Project Status |
Completed (Fiscal Year 2020)
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| Budget Amount *help |
¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000)
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| Keywords | 電子顕微鏡 / Cryo-fixation / 急速高圧固定 / スパイン / グリア細胞 / 高圧凍結固定法 / FIB/SEM / 高圧クライオ固定 / 三次元微細構造 / 細胞・組織 |
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| Outline of Final Research Achievements |
In this study in order to understand the native ultrastructure of enteric glial cells with novel three-dimensional electron microscopy, such as the Focused ion beam / Scanning electron microscopy (FIB/SEM), a snap freezing method of liquid nitrogen jets, combined with very high pressures (cryo-fixation), was analyzed. At first, with using the mouse cortex tissues, the difference among the cryo-fixation and conventional chemical-fixations was confirmed. As a result, dendritic spines showed significantly thinner necks with cryo-fixation. Ｔhis work highlights the usefulness of cryo-fixation for obtaining a more native and detailed view of the architecture of cells and tissues.
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| Academic Significance and Societal Importance of the Research Achievements |
新しい電子顕微鏡技術の発展に伴い、これまでは不可能であった電子顕微鏡像による定量的評価が可能となった。そのような中、従来の電顕用組織固定法である強いアルデヒドを用いた化学固定では、細胞膜の構造は明瞭に維持されるものの、細胞や組織本来の形態が消失する可能性が特に強く危惧される。本研究で検討した化学固定液を用いない急速高圧凍結固定法Cryo-fixationは、細胞や組織のより本来の姿に近い形態を維持するものと考えられ、今後のあらゆる生理機能の根幹をなす形態解析において重要な成果である。
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Report
(4 results)
Research Products
(10 results)
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[Journal Article] Astrocytic phagocytosis is a compensatory mechanism for microglial dysfunction2020
Author(s)
Konishi Hiroyuki、Okamoto Takayuki、Hara Yuichiro、Komine Okiru、Tamada Hiromi、Maeda Mitsuyo、Osako Fumika、Kobayashi Masaaki、Nishiyama Akira、Kataoka Yosky、Takai Toshiyuki、Udagawa Nobuyuki、Jung Steffen、Ozato Keiko、Tamura Tomohiko、Tsuda Makoto、Yamanaka Koji、Ogi Tomoo、Sato Katsuaki、Kiyama Hiroshi
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Journal Title
The EMBO Journal
Volume: 39
Issue: 22
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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