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Pré-Publication, Document De Travail Année : 2020

P2X-GCaMPs as versatile tools for imaging extracellular ATP signaling

Matthias Ollivier
  • Fonction : Auteur
Juline Beudez
  • Fonction : Auteur
Nathalie Linck
  • Fonction : Auteur
Vincent Compan
Francois Rassendren

Résumé

Adenosine 5’ triphosphate (ATP) is an extracellular signaling molecule involved in numerous physiological and pathological processes. Yet, in situ characterization of the spatiotemporal dynamic of extracellular ATP is still challenging due to the lack of sensor with appropriate specificity, sensitivity and kinetics. Here we report the development of biosensors based on the fusion of cation permeable ATP receptors (P2X) to genetically encoded calcium sensors (GECI). By combining the features of P2X receptors with the high signal to noise ratio of GECIs, we generated ultrasensitive green and red fluorescent sniffers that detect nanomolar ATP concentrations in situ and also enable the tracking of P2X receptor activity. We provide the proof of concept that these sensors can dynamically track ATP release evoked by neuronal depolarization or by extracellular hypotonicity. Targeting these P2X-based biosensors to diverse cell types should advance our knowledge of extracellular ATP dynamics in vivo .

Domaines

Neurobiologie

Dates et versions

hal-02989722 , version 1 (05-11-2020)

Identifiants

Citer

Matthias Ollivier, Juline Beudez, Nathalie Linck, Thomas Grutter, Vincent Compan, et al.. P2X-GCaMPs as versatile tools for imaging extracellular ATP signaling. 2020. ⟨hal-02989722⟩
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