GFP-Aequorin Protein Sensor for Ex Vivo and In Vivo Imaging of Ca 2+ Dynamics in High-Ca 2+ Organelles

Abstract : Proper functioning of organelles such as the ER or the Golgi apparatus requires luminal accumulation of Ca(2+) at high concentrations. Here we describe a ratiometric low-affinity Ca(2+) sensor of the GFP-aequorin protein (GAP) family optimized for measurements in high-Ca(2+) concentration environments. Transgenic animals expressing the ER-targeted sensor allowed monitoring of Ca(2+) signals inside the organelle. The use of the sensor was demonstrated under three experimental paradigms: (1) ER Ca(2+) oscillations in cultured astrocytes, (2) ex vivo functional mapping of cholinergic receptors triggering ER Ca(2+) release in acute hippocampal slices from transgenic mice, and (3) in vivo sarcoplasmic reticulum Ca(2+) dynamics in the muscle of transgenic flies. Our results provide proof of the suitability of the new biosensors to monitor Ca(2+) dynamics inside intracellular organelles under physiological conditions and open an avenue to explore complex Ca(2+) signaling in animal models of health and disease.
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https://hal-univ-evry.archives-ouvertes.fr/hal-02179235
Contributor : Paloma Navas Navarro <>
Submitted on : Wednesday, July 10, 2019 - 3:36:06 PM
Last modification on : Wednesday, July 10, 2019 - 4:06:45 PM

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Paloma Navas-Navarro, Jonathan Rojo-Ruiz, Macarena Rodríguez-Prados, María Dolores Ganfornina, Loren Looger, et al.. GFP-Aequorin Protein Sensor for Ex Vivo and In Vivo Imaging of Ca 2+ Dynamics in High-Ca 2+ Organelles. Cell Chemical Biology, Cell Press, 2016, 23 (6), pp.738-745. ⟨10.1016/j.chembiol.2016.05.010⟩. ⟨hal-02179235⟩

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