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RGB‐Color Intensiometric Indicators to Visualize Spatiotemporal Dynamics of ATP in Single Cells
Authors:Dr Satoshi Arai  Dr Rókus Kriszt  Kazuki Harada  Dr Liang‐Sheng Looi  Shogo Matsuda  Devina Wongso  Dr Satoshi Suo  Prof Shoichi Ishiura  Prof Yu‐Hua Tseng  Prof Michael Raghunath  Prof Toshiro Ito  Prof Takashi Tsuboi  Prof Tetsuya Kitaguchi
Institution:1. Cell Signaling Group, Waseda Bioscience Research Institute in Singapore (WABIOS), Singapore, Republic of Singapore;2. Organization for University Research Initiatives, Waseda University, Tokyo, Japan;3. AMED-PRIME (Japan) Agency for Medical Research and Development, Tokyo, Japan;4. Department of Biomedical Engineering, National University of Singapore, Republic of Singapore;5. Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Tokyo, Japan;6. Temasek Life Sciences Laboratory, National University of Singapore, Singapore, Republic of Singapore;7. Department of Biological Sciences, National University of Singapore, Singapore, Republic of Singapore;8. Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan;9. Section on Integrative Physiology and Metabolism, Joslin Diabetes Center, Harvard Medical School, Boston, MA, USA;10. Current Address: Laboratory for Chemistry and Life Science, Institute of Innovative Research, Tokyo Institute of Technology, Kanagawa, Japan
Abstract: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.
Keywords:ATP  brown adipocytes  fluorescent probes  live cell imaging  protein engineering
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