RGB‐Color Intensiometric Indicators to Visualize Spatiotemporal Dynamics of ATP in Single Cells

• Published in: Angewandte Chemie International Edition Vol. 57; no. 34; pp. 10873 - 10878
• Main Authors: Arai, Satoshi, Kriszt, Rókus, Harada, Kazuki, Looi, Liang‐Sheng, Matsuda, Shogo, Wongso, Devina, Suo, Satoshi, Ishiura, Shoichi, Tseng, Yu‐Hua, Raghunath, Michael, Ito, Toshiro, Tsuboi, Takashi, Kitaguchi, Tetsuya
• Format: Journal Article
• Language: English
• Published: Germany Wiley Subscription Services, Inc 20.08.2018
• Edition: International ed. in English
• Subjects: Adenosine triphosphate; Adenosine Triphosphate - metabolism; Adipocytes, Brown - cytology; Adipocytes, Brown - metabolism; Animals; ATP; brown adipocytes; Calcium; Calcium - metabolism; Calcium ions; Cells, Cultured; Color; Cyclic AMP - metabolism; Cytoplasm; Cytoplasm - metabolism; Dynamics; Energy metabolism; Fluorescence; Fluorescent indicators; fluorescent probes; Glycolysis; HeLa Cells; Humans; Hydrogen-Ion Concentration; Indicators; Intracellular signalling; live cell imaging; Luminescent Proteins - metabolism; Metabolism; Mice; Mitochondria; Mitochondria - metabolism; Organelles; Oxidative Phosphorylation; Plant cells; protein engineering; Proteins; Single-Cell Analysis; protein engineering; live cell imaging; brown adipocytes; ATP; fluorescent probes
• ISSN: 1433-7851; 1521-3773; 1521-3773
• DOI: 10.1002/anie.201804304

Adenosine triphosphate (ATP) provides energy for the regulation of multiple cellular processes in living organisms. Capturing the spatiotemporal dynamics of ATP in single cells is fundamental to our understanding of the mechanisms underlying cellular energy metabolism. However, it has remained challenging to visualize the dynamics of ATP in and between distinct intracellular organelles and its interplay with other signaling molecules. Using single fluorescent proteins, multicolor ATP indicators were developed, enabling the simultaneous visualization of subcellular ATP dynamics in the cytoplasm and mitochondria of cells derived from mammals, plants, and worms. Furthermore, in combination with additional fluorescent indicators, the dynamic interplay of ATP, cAMP, and Ca2+ could be visualized in activated brown adipocyte. This set of indicator tools will facilitate future research into energy metabolism. A multicolor palette of fluorescent indicators is beneficial in biology and biochemistry. Red, green, and blue‐colored genetically encoded indicators were designed that are capable of reporting changes in ATP concentration as a change in single fluorescence emission. The expanded palette enables simultaneous imaging of ATP dynamics in the cytoplasm versus mitochondria, which has not been achieved with other techniques.