Department of Neurobiology, Harvard Medical School, 220 Longwood Avenue, Boston, Massachusetts 02115, USA.
Nat Commun. 2013;4:2550. doi: 10.1038/ncomms3550.
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.
ADP 比率是细胞能量状态的一个关键参数，调节许多代谢活动。在这里，我们报告了一种优化的基因编码荧光生物传感器 PercevalHR，它可以感知 ATP:ADP 比率。PercevalHR 被调整到哺乳动物细胞中预期的细胞内 ATP:ADP 范围，并且可以在活样本中使用单光子或双光子显微镜进行检测。我们使用 PercevalHR 来可视化神经元暴露于多种刺激时 ATP:ADP 的活性依赖性变化，证明它是能量消耗和产生生理变化的敏感报告者。我们还使用 PercevalHR 来可视化细胞内的 ATP:ADP，同时记录单个细胞中 ATP 敏感钾 (KATP) 通道的电流，表明 PercevalHR 可以在完整细胞中研究 ATP:ADP 和 KATP 通道开放概率的协调变化。由于其能够在几秒钟内监测细胞能量状态的变化，PercevalHR 应该成为代谢研究的多功能工具。
