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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >GAP, an aequorin-based fluorescent indicator for imaging Ca~(2+) in organelles
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GAP, an aequorin-based fluorescent indicator for imaging Ca~(2+) in organelles

机译：GAP, an aequorin-based fluorescent indicator for imaging Ca~(2+) in organelles

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摘要

Genetically encoded calcium indicators allow monitoring subcellular Ca~(2+) 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 Ca~(2+) 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 Mg~(2+), tunable Ca~(2+) 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 Ca~(2+) indicators for organelles and becomes a valuable tool for in vivo Ca~(2+) imaging applications.

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