Analysis of neuro-immune synapse by fluorescence and bioluminescence imaging
| Project/Area Number |
17K07374
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Biophysics
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| Research Institution | Aichi Gakuin University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
鈴木 崇弘 愛知学院大学, 歯学部, 教授 (70298545)
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| Project Period (FY) |
2017-04-01 – 2020-03-31
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| Project Status |
Completed (Fiscal Year 2019)
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| Budget Amount *help |
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
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| Keywords | イメージング / マスト細胞 / 脱顆粒 / 接着分子 / 細胞骨格 / 細胞外マトリックス / 発光イメージング / 開口放出 / 細胞間相互作用 / 生物物理 / 細胞情報 |
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| Outline of Final Research Achievements |
We developed a method of video-rate bioluminescence imaging to directly detect degranulation from a single mast cell by measuring luminescence activity derived from the enzymatic reaction between Gaussia luciferase (GLase) and its substrate coelenterazine. Bioluminescence imaging analysis of mast cells (MCs) expressing NPY-GLase showed that the luminescence signals of the secreted NPY-GLase were repeatedly detected after the addition of an antigen.
In addition, we found that an antigen-induced increase in intracellular Ca2+ concentration was suppressed slightly in cells on hydrogel-coated dishes compared with those on non-coated dishes, whereas their subsequent degranulation was largely inhibited in cells adherent to the hydrogel. Vinculin was distributed in a dot-like manner at the bottom of resting cells on non-coated dishes but not on hydrogel-coated dishes. Moreover, microtubule reorganization and acetylation were also suppressed in activated MCs adherent to the hydrogel.
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| Academic Significance and Societal Importance of the Research Achievements |
神経－免疫シナプスの分子機構の解析に必要な基盤技術を確立することができた。生体内の様々な組織に分布しているマスト細胞の多くは、神経細胞の近傍に存在していることが知られている。また、両細胞は組織ごとに異なる細胞外マトリックス（ECM）に接着して生存している。また、ECMの性質は疾患の進行に伴う組織の繊維化や、マスト細胞の分泌するケミカルメディエーターによって変化することが知られている。今後、本研究の成果を活用して、神経細胞とマスト細胞の相互作用の研究を進めていくことにより、新たな視点から抗原非依存的な炎症反応誘導の分子機構を明らかにしていくことが可能であると考えている。
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Report
(4 results)
Research Products
(18 results)
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[Journal Article] Video-rate bioluminescence imaging of degranulation of mast cells attached to the extracellular matrix.2018
Author(s)
Yokawa, S., Suzuki, T., Hayashi, A., Inouye, S., Inoh, Y., Furuno, T.
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Journal Title
Front. Cell Dev. Biol.
Volume: 6
Pages: 74-74
DOI
Related Report
Peer Reviewed / Open Access
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