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Title

GAP, an aequorin-based fluorescent indicator for imaging Ca2+ in organelles

AuthorsRodríguez-García, Arancha ; Rojo-Ruiz, Jonathan; Navas-Navarro, Paloma; Aulestia, Francisco J.; Gallego‑Sandín, Sonia ; García-Sancho, Javier ; Alonso, María Teresa
KeywordsGolgi apparatus
Hippocampus
Motor neuron
Calcium signalling
Issue Date2014
PublisherNational Academy of Sciences (U.S.)
CitationProceedings of the National Academy of Sciences 111(7): 2584-2589 (2014)
AbstractGenetically encoded calcium indicators allow monitoring subcellular Ca2+ signals inside organelles. Most genetically encoded calcium indicators are fusions of endogenous calcium-binding proteins whose functionality in vivo may be perturbed by competition with cellular partners.We describe here a novel family of fluorescent Ca2+ sensors based on the fusion of two Aequorea victoria proteins, GFP and apo-aequorin (GAP). GAP exhibited a unique combination of features: dual-excitation ratiometric imaging, high dynamic range, good signal-to-noise ratio, insensitivity to pH and Mg2+, tunable Ca2+ affinity, uncomplicated calibration, and targetability to five distinct organelles. Moreover, transgenic mice for endoplasmic reticulum-targeted GAP exhibited a robust long-term expression that correlated well with its reproducible performance in various neural tissues. This biosensor fills a gap in the actual repertoire of Ca2+ indicators for organelles and becomes a valuable tool for in vivo Ca2+ imaging applications.
URIhttp://hdl.handle.net/10261/116931
DOI10.1073/pnas.1316539111
Identifiersdoi: 10.1073/pnas.1316539111
e-issn: 1091-6490
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