The purinergic transmitter ATP (adenosine 5′-triphosphate) plays an essential role in both the central and peripheral nervous systems, and the ability to directly measure extracellular ATP in real time will increase our understanding of its physiological functions. Here, we developed a sensitive GPCR activation-based ATP sensor called GRAB_ATP1.0, with a robust fluorescence response to extracellular ATP when expressed in several cell types. This sensor has sub-second kinetics, has ATP affinity in the range of tens of nanomolar, and can be used to localize ATP release with subcellular resolution. Using this sensor, we monitored ATP release under a variety of in vitro and in vivo conditions, including stimuli-induced and spontaneous ATP release in primary hippocampal cultures, injury-induced ATP release in a zebrafish model, and lipopolysaccharides-induced ATP-release events in individual astrocytes in the mouse cortex. Thus, the GRAB_ATP1.0 sensor is a sensitive, versatile tool for monitoring ATP release and dynamics under both physiological and pathophysiological conditions.

嘌呤能递质 ATP（腺苷 5'-三磷酸）在中枢神经系统和外周神经系统中都发挥着重要作用，实时直接测量细胞外 ATP 的能力将增加我们对其生理功能的理解。在这里，我们开发了一种敏感的基于 GPCR 激活的 ATP 传感器，称为 GRAB _ATP1.0，当在几种细胞类型中表达时，它对细胞外 ATP 具有强大的荧光响应。该传感器具有亚秒级动力学，具有数十纳摩尔范围内的 ATP 亲和力，可用于以亚细胞分辨率定位 ATP 释放。使用该传感器，我们监测了各种体外和体内条件下的 ATP 释放条件，包括原代海马培养物中刺激诱导和自发的 ATP 释放，斑马鱼模型中损伤诱导的 ATP 释放，以及小鼠皮层中单个星形胶质细胞中脂多糖诱导的 ATP 释放事件。因此，GRAB _ATP1.0传感器是一种灵敏的多功能工具，用于监测生理和病理生理条件下的 ATP 释放和动态。