2018 Fiscal Year Final Research Report

In vivo imaging with FRET mice

Research Project

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Project/Area Number	15H02397
Research Category	Grant-in-Aid for Scientific Research (A)
Allocation Type	Single-year Grants
Section	一般
Research Field	Cell biology
Research Institution	Kyoto University
Principal Investigator	Matsuda Michiyuki 京都大学, 生命科学研究科, 教授 (10199812)
Co-Investigator(Kenkyū-buntansha)	平塚拓也京都大学, 医学(系)研究科(研究院), 助教 (90641639)
Research Collaborator	HIRATSUKA Takuya AOKI Kazuhiro IMAjo Masamichi KOMATSU Naoki SUMIYAMA Kenta TSUKIJI Shinya IMAYOSHI Itaru TERAI Kenta HOTTA Kazuhiro MURATA Tomokazu SATO Masaya
Project Period (FY)	2015-04-01 – 2019-03-31
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
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.
Free Research Field	細胞生物学
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。