| Project/Area Number |
15H02397
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Cell biology
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| Research Institution | Kyoto University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
平塚 拓也 京都大学, 医学(系)研究科(研究院), 助教 (90641639)
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| Research Collaborator |
HIRATSUKA Takuya
AOKI Kazuhiro
IMAjo Masamichi
KOMATSU Naoki
SUMIYAMA Kenta
TSUKIJI Shinya
IMAYOSHI Itaru
TERAI Kenta
HOTTA Kazuhiro
MURATA Tomokazu
SATO Masaya
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| Project Period (FY) |
2015-04-01 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2018)
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| Budget Amount *help |
¥39,910,000 (Direct Cost: ¥30,700,000、Indirect Cost: ¥9,210,000)
Fiscal Year 2018: ¥9,750,000 (Direct Cost: ¥7,500,000、Indirect Cost: ¥2,250,000)
Fiscal Year 2017: ¥9,750,000 (Direct Cost: ¥7,500,000、Indirect Cost: ¥2,250,000)
Fiscal Year 2016: ¥9,750,000 (Direct Cost: ¥7,500,000、Indirect Cost: ¥2,250,000)
Fiscal Year 2015: ¥10,660,000 (Direct Cost: ¥8,200,000、Indirect Cost: ¥2,460,000)
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| Keywords | 蛍光プローブ / 組織形態マーカー / FRETバイオセンサー / 機械学習 / ROCK / 蛍光イメージング / 血管収縮 / イメージング / 透明化 / FRET / トランスジェニックマウス / ERK MAPキナーゼ / 細胞増殖 / 蛍光タンパク質 / 多光子顕微鏡 |
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| Outline of Final Research Achievements |
Hematoxylin and eosin (H&E) staining has been the de-facto standard for histological studies. We have developed a genetically encoded fluorescent marker, NuCyM, which is designed to recapitulate H&E staining patterns in vivo. We generated a transgenic mouse line ubiquitously expressing NyCyM. NuCyM evenly marked the plasma membrane, cytoplasm and nucleus in most tissues. In the NuCyM-expressing cells, cell division of a single cell was clearly observed as five basic phases during M phase by three-dimensional imaging. We next crossed NuCyM mice with transgenic mice expressing an ERK biosensor. Using NuCyM, ERK activity in each cell could be extracted from the FRET images. To further accelerate the image analysis, we employed machine learning-based segmentation methods, and thereby automatically quantitated ERK activity in each cell. In conclusion, NuCyM is a versatile cell morphological marker that enables us to grasp histological information as with H&E staining.
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| Academic Significance and Societal Importance of the Research Achievements |
生物学の研究は、培養皿上のクローン化された細胞から、オルガノイド、さらには生体組織へとその中心を移しつつある。今回開発したNuCyMは蓄積された組織学の叡智を最新の分子プローブへと繋ぐツールである。今後の病態解明あるいは創薬研究に役立つツールとなることが期待される。また、機械学習を用いた細胞分別法はさまざまに開発されつつあるが、機械学習を念頭においた組織染色あるいは細胞染色法が必要であり、本研究はその嚆矢としても重要である。
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