Details

Autor(en) / Beteiligte

• Wu, Zhaofa
• He, Kaikai
• Chen, Yue
• Li, Hongyu
• Pan, Sunlei
• Li, Bohan
• Liu, Tingting
• Xi, Fengxue
• Deng, Fei
• Wang, Huan
• Du, Jiulin
• Jing, Miao
• Li, Yulong

Titel

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

Ist Teil von

• Neuron (Cambridge, Mass.), 2022-03, Vol.110 (5), p.770-782.e5

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2022

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• 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. [Display omitted] •GRABATP is a genetically encoded GPCR activation-based sensor for extracellular ATP•GRABATP can detect ATP with high sensitivity and spatiotemporal resolution•GRABATP reveals injury-induced in vivo propagation of ATP in zebrafish larvae•GRABATP reveals LPS-induced local ATP-release events in the mouse cortex in vivo Wu et al. developed a sensitive GPCR activation-based ATP sensor, GRABATP1.0, for monitoring extracellular ATP dynamics under a variety of conditions. In zebrafish, GRABATP1.0 detected brain injury-induced in vivo propagation of ATP. In mouse cortex, GRABATP1.0 revealed ATP-releasing events at the resolution of a single astrocyte after LPS-induced inflammation.