development of a method to analyze local organelle function in situ

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K22509
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 48:Biomedical structure and function and related fields
• Research Institution: University of Fukui
• Principal Investigator: Matsuoka Satoshi 福井大学, 学術研究院医学系部門, 教授 (00263096)
• Co-Investigator(Kenkyū-buntansha): 竹内 綾子 福井大学, 学術研究院医学系部門, 准教授 (00378704)
• Project Period (FY): 2019-06-28 – 2022-03-31
• Keywords: ミトコンドリア / 膜電位 / 微小電極

Outline of Final Research Achievements

There is no way to measure directly mitochondrial membrane potential in living cells. We aimed to develop a new micropipette method to measure directly mitochondrial membrane potential. After the improvement of experimental equipment, selection of proper glass capillary, adjustment of puller and selection of proper mitochondria-specific fluorescence dye, we tried direct measurement of mitochondrial membrane potential in living cardiomyocytes with microelectrodes. However, a large negative membrane potential corresponding to mitochondrial membrane potential could not be detected. The possible reasons are damage and depolarization of mitochondria by the microelectrodes, or non-penetration of microelectrodes through mitochondrial inner/outer membrane. It seems, however, possible to measure directly mitochondrial membrane potential with cells having giant mitochondria.

Free Research Field

生理学

Academic Significance and Societal Importance of the Research Achievements

ミトコンドリア膜電位の直接測定を可能にするために行った測定機器の改良、ガラス電極の選定、電極作成装置の調整、ミトコンドリア染色蛍光色素の選定等の情報は、今後の展開への貴重な資料となる。また、より大きなミトコンドリアを有する細胞を用いることで、生細胞のミトコンドリア膜電位の直接測定は可能であるとの見通しが立った点は意義がある。