Development of small fluorescent probes for the analysis of in vivo autophagy kinetics

2020 Fiscal Year Final Research Report

• Project/Area Number: 19K16119
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 44010:Cell biology-related
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: Sakurai Hajime 東京医科歯科大学, 難治疾患研究所, プロジェクト助教 (00796732)
• Project Period (FY): 2019-04-01 – 2021-03-31
• Keywords: オートファジー / 蛍光プローブ

Outline of Final Research Achievements

Studies of various autophagy-deficient mice have revealed the essential roles in many aspects. However, "when/where is autophagy activated in vivo?" is still almost uncovered. The fluorescent probes which detect autophagic activities without transgenic processes are needed to approach these issues. Even though several probes are commercially available, the characterization of these probes are hidden so far.
Recently, we developed the new green-fluorescent probes for monitoring autophagy. In this study, we further developed a new red-fluorescent probe. Then, we showed combinational labeling with these green and red probes permits monitoring autophagic flux, since we analyzed the labeling nature of these probes in detail. Furthermore, the developed new probe makes it possible to detect autophagic activity in vivo.

Free Research Field

細胞生物学

Academic Significance and Societal Importance of the Research Achievements

本研究で開発した3つの蛍光プローブが，各々オートファジー現象のどのステージを標識できるのかを詳細に解明し，それぞれ異なるステージを標識し分けることが可能であることを明らかにした．この成果により，関連遺伝子の解析や薬剤の開発において，開発したプローブを活用することで，オートファジー現象のどのステージに着目すればよいのかを簡便に知ることが可能となった．
また，遺伝子導入を必要とせずに生体内でのオートファジー活性を可視化できるため，原因未知の疾患とオートファジーとの関連性の解明に役立てることが期待できる．