First Author | Rodriguez-Garcia A | Year | 2014 |
Journal | Proc Natl Acad Sci U S A | Volume | 111 |
Issue | 7 | Pages | 2584-9 |
PubMed ID | 24501126 | Mgi Jnum | J:206829 |
Mgi Id | MGI:5553021 | Doi | 10.1073/pnas.1316539111 |
Citation | Rodriguez-Garcia A, et al. (2014) GAP, an aequorin-based fluorescent indicator for imaging Ca2+ in organelles. Proc Natl Acad Sci U S A 111(7):2584-9 |
abstractText | 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. |