2024-03-29T02:38:34Zhttps://uvadoc.uva.es/oai/requestoai:uvadoc.uva.es:10324/351932021-06-24T07:36:21Zcom_10324_32522com_10324_952com_10324_894col_10324_32523
GFP-Aequorin Protein Sensor for Ex Vivo and In Vivo Imaging of Ca2+ Dynamics in High-Ca2+ Organelles
Navas Navarro, Paloma
Rojo Ruiz, Jonathan
Rodríguez Prados, Macarena
Ganfornina Álvarez, María Dolores
Looger, Loren L.
Alonso Alonso, María Teresa
García-Sancho Martín, Francisco Javier
Producción Científica
Proper functioning of organelles such as the ER or the Golgi apparatus requires luminal accumulation of Ca2+ at high concentrations. Here we describe a ratiometric low-affinity Ca2+ sensor of the GFP-aequorin protein (GAP) family optimized for measurements in high-Ca2+ concentration environments. Transgenic animals expressing the ER-targeted sensor allowed monitoring of Ca2+ signals inside the organelle. The use of the sensor was demonstrated under three experimental paradigms: (1) ER Ca2+ oscillations in cultured astrocytes, (2) ex vivo functional mapping of cholinergic receptors triggering ER Ca2+ release in acute hippocampal slices from transgenic mice, and (3) in vivo sarcoplasmic reticulum Ca2+ dynamics in the muscle of transgenic flies. Our results provide proof of the suitability of the new biosensors to monitor Ca2+ dynamics inside intracellular organelles under physiological conditions and open an avenue to explore complex Ca2+ signaling in animal models of health and disease.
2019-03-22T11:17:49Z
2019-03-22T11:17:49Z
2016
info:eu-repo/semantics/article
Cell Chemical Biology, 2016, Volume 23, Issue 6, Pages 738-745
2451-9456
http://uvadoc.uva.es/handle/10324/35193
https://doi.org/10.1016/j.chembiol.2016.05.010
eng
https://www.sciencedirect.com/science/article/pii/S2451945616301635
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Elsevier
SI