2. Academic Validation
  3. A sensitive GRAB sensor for detecting extracellular ATP in vitro and in vivo

A sensitive GRAB sensor for detecting extracellular ATP in vitro and in vivo

  • Neuron. 2022 Mar 2;110(5):770-782.e5. doi: 10.1016/j.neuron.2021.11.027.
Zhaofa Wu 1,  Kaikai He 2,  Yue Chen 3,  Hongyu Li 4,  Sunlei Pan 5,  Bohan Li 2,  Tingting Liu 4,  Fengxue Xi 6,  Fei Deng 2,  Huan Wang 2,  Jiulin Du 4,  Miao Jing 3,  Yulong Li 7
Affiliations

Affiliations

  • 1 State Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing 100871, China; PKU-IDG/McGovern Institute for Brain Research, Beijing 100871, China. Electronic address: wuzhaofa@pku.edu.cn.
  • 2 State Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing 100871, China; PKU-IDG/McGovern Institute for Brain Research, Beijing 100871, China.
  • 3 Chinese Institute for Brain Research, Beijing 102206, China.
  • 4 Institute of Neuroscience, State Key Laboratory of Neuroscience, Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, 320 Yue-Yang Road, Shanghai 200031, China.
  • 5 State Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing 100871, China; PKU-IDG/McGovern Institute for Brain Research, Beijing 100871, China; Peking-Tsinghua Center for Life Sciences, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing 100871, China.
  • 6 Chinese Institute for Brain Research, Beijing 102206, China; School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China.
  • 7 State Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing 100871, China; PKU-IDG/McGovern Institute for Brain Research, Beijing 100871, China; Peking-Tsinghua Center for Life Sciences, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing 100871, China; Chinese Institute for Brain Research, Beijing 102206, China. Electronic address: yulongli@pku.edu.cn.
PMID: 34942116 DOI: 10.1016/j.neuron.2021.11.027
Abstract

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 GRABATP1.0, 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 GRABATP1.0 sensor is a sensitive, versatile tool for monitoring ATP release and dynamics under both physiological and pathophysiological conditions.

Keywords

ATP; GPCR; GRAB; fluorescent sensors; genetically encoded; imaging; injury; neuroinflammation; purinergic signaling.

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