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Imaging energy status in live cells with a fluorescent biosensor of the intracellular ATP-to-ADP ratio

Author

Listed:
  • Mathew Tantama

    (Harvard Medical School)

  • Juan Ramón Martínez-François

    (Harvard Medical School)

  • Rebecca Mongeon

    (Harvard Medical School)

  • Gary Yellen

    (Harvard Medical School)

Abstract

The ATP:ADP ratio is a critical parameter of cellular energy status that regulates many metabolic activities. Here we report an optimized genetically encoded fluorescent biosensor, PercevalHR, that senses the ATP:ADP ratio. PercevalHR is tuned to the range of intracellular ATP:ADP expected in mammalian cells, and it can be used with one- or two-photon microscopy in live samples. We use PercevalHR to visualize activity-dependent changes in ATP:ADP when neurons are exposed to multiple stimuli, demonstrating that it is a sensitive reporter of physiological changes in energy consumption and production. We also use PercevalHR to visualize intracellular ATP:ADP while simultaneously recording currents from ATP-sensitive potassium (KATP) channels in single cells, showing that PercevalHR enables the study of coordinated variation in ATP:ADP and KATP channel open probability in intact cells. With its ability to monitor changes in cellular energetics within seconds, PercevalHR should be a versatile tool for metabolic research.

Suggested Citation

  • Mathew Tantama & Juan Ramón Martínez-François & Rebecca Mongeon & Gary Yellen, 2013. "Imaging energy status in live cells with a fluorescent biosensor of the intracellular ATP-to-ADP ratio," Nature Communications, Nature, vol. 4(1), pages 1-11, December.
    • Handle: RePEc:nat:natcom:v:4:y:2013:i:1:d:10.1038_ncomms3550
    DOI: 10.1038/ncomms3550
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    Cited by:

    1. Blanca Jiménez-Gómez & Patricia Ortega-Sáenz & Lin Gao & Patricia González-Rodríguez & Paula García-Flores & Navdeep Chandel & José López-Barneo, 2023. "Transgenic NADH dehydrogenase restores oxygen regulation of breathing in mitochondrial complex I-deficient mice," Nature Communications, Nature, vol. 14(1), pages 1-14, December.

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