Alonso María Teresa, Rodríguez-Prados Macarena, Navas-Navarro Paloma, Rojo-Ruiz Jonathan, García-Sancho Javier
Instituto de Biología y Genética Molecular (IBGM), Universidad de Valladolid and Consejo Superior de Investigaciones Científicas (CSIC), c/Sanz y Forés 3, 47003 Valladolid, Spain.
Instituto de Biología y Genética Molecular (IBGM), Universidad de Valladolid and Consejo Superior de Investigaciones Científicas (CSIC), c/Sanz y Forés 3, 47003 Valladolid, Spain.
Cell Calcium. 2017 Jun;64:3-11. doi: 10.1016/j.ceca.2017.01.006. Epub 2017 Jan 16.
Aequorins are excellent tools for measuring intra-organellar Ca and assessing its role in physiological and pathological functions. Here we review targeting strategies to express aequorins in various organelles. We address critical topics such as probe affinity tuning as well as normalization and calibration of the signal. We also focus on bioluminescent Ca imaging in nucleus or mitochondria of living cells. Finally, recent advances with a new chimeric GFP-aequorin protein (GAP), which can be used either as luminescent or fluorescent Ca probe, are presented. GAP is robustly expressed in transgenic flies and mice, where it has proven to be a suitable Ca indicator for monitoring physiological Ca signaling ex vivo and in vivo.
水母发光蛋白是用于测量细胞器内钙离子浓度并评估其在生理和病理功能中作用的优秀工具。在此，我们综述了在各种细胞器中表达水母发光蛋白的靶向策略。我们讨论了诸如探针亲和力调节以及信号归一化和校准等关键主题。我们还专注于活细胞细胞核或线粒体中的生物发光钙成像。最后，介绍了一种新型嵌合绿色荧光蛋白-水母发光蛋白（GAP）的最新进展，它既可以用作发光钙探针，也可以用作荧光钙探针。GAP在转基因果蝇和小鼠中能高效表达，已证明它是一种适用于在体外和体内监测生理钙信号的钙指示剂。
