PUBLICATION

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

Authors
Wu, Z., He, K., Chen, Y., Li, H., Pan, S., Li, B., Liu, T., Xi, F., Deng, F., Wang, H., Du, J., Jing, M., Li, Y.
ID
ZDB-PUB-211224-28
Date
2021
Source
Neuron   110(5): 770-782.e5 (Journal)
Registered Authors
Du, Jiu Lin, Li, Hong-Yu
Keywords
ATP, GPCR, GRAB, fluorescent sensors, genetically encoded, imaging, injury, neuroinflammation, purinergic signaling
MeSH Terms
  • Adenosine Triphosphate*/metabolism
  • Animals
  • Astrocytes/metabolism
  • Lipopolysaccharides/pharmacology
  • Mice
  • Zebrafish*
PubMed
34942116 Full text @ Neuron
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.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping