[HTML][HTML] Imaging extracellular ATP with a genetically-encoded, ratiometric fluorescent sensor

JM Conley, S Radhakrishnan, SA Valentino… - PLoS …, 2017 - journals.plos.org
JM Conley, S Radhakrishnan, SA Valentino, M Tantama
PLoS One, 2017journals.plos.org
Extracellular adenosine triphosphate (ATP) is a key purinergic signal that mediates cell-to-
cell communication both within and between organ systems. We address the need for a
robust and minimally invasive approach to measuring extracellular ATP by re-engineering
the ATeam ATP sensor to be expressed on the cell surface. Using this approach, we image
real-time changes in extracellular ATP levels with a sensor that is fully genetically-encoded
and does not require an exogenous substrate. In addition, the sensor is ratiometric to allow …
Extracellular adenosine triphosphate (ATP) is a key purinergic signal that mediates cell-to-cell communication both within and between organ systems. We address the need for a robust and minimally invasive approach to measuring extracellular ATP by re-engineering the ATeam ATP sensor to be expressed on the cell surface. Using this approach, we image real-time changes in extracellular ATP levels with a sensor that is fully genetically-encoded and does not require an exogenous substrate. In addition, the sensor is ratiometric to allow for reliable quantitation of extracellular ATP fluxes. Using live-cell microscopy, we characterize sensor performance when expressed on cultured Neuro2A cells, and we measure both stimulated release of ATP and its clearance by ectonucleotidases. Thus, this proof-of-principle demonstrates a first-generation sensor to report extracellular ATP dynamics that may be useful for studying purinergic signaling in living specimens.
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