Using aequorin probes to measure Ca2+ in intracellular organelles. (June 2017)

• Record Type: Journal Article
• Authors: Alonso, María Teresa; Rodríguez-Prados, Macarena; Navas-Navarro, Paloma; Rojo-Ruiz, Jonathan; García-Sancho, Javier
• Is Part Of: Cell calcium. Volume 64(2017)
• Page Start: 3
• Page End: 11
• Publication Date: 2017-06
• Subjects: ER endoplasmic reticulum -- SR sarcoplasmic reticulum -- AEQ aequorin -- [Ca2+]C, [Ca2+]N, [Ca2+]M, and [Ca2+]ER Ca2+ concentration in cytosol, nucleus, mitochondria and endoplasmic reticulum, respectively -- GECI genetically encoded Ca2+ indicator -- KDEL tetrapeptide with sequence Lys-Asp-Glu-Leu -- InsP3 inositol (1, 4, 5)-trisphosphate -- SERCA sarcoendoplasmic reticulum Ca2+-ATPase -- SPCA1 secretory pathway Ca2+ ATPase 1; Calcium signaling -- Aequorin -- Intracellular organella -- Endoplasmic reticulum -- Golgi apparatus -- Hippocampus -- Skeletal muscle; Calcium -- Metabolism -- Periodicals; Vertebrates -- Physiology -- Periodicals; Calcium -- Physiological effect -- Periodicals; Cell physiology -- Periodicals; Calcium in the body -- Periodicals; 572.516
• DOI: 10.1016/j.ceca.2017.01.006
• Languages: English
• ISSNs: 0143-4160

Abstract

Graphical abstract: Highlights: Aequorins are valuable orthogonal tools for measuring Ca 2+ levels in organelles. GFP-aequorin fusions can be used both as luminescent and as fluorescent probes. Transgenic animals show normal phenotype and can be used for in vivo Ca 2+ monitoring. Abstract: Aequorins are excellent tools for measuring intra-organellar Ca 2+ 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 2+ 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 2+ probe, are presented. GAP is robustly expressed in transgenic flies and mice, where it has proven to be a suitable Ca 2+ indicator for monitoring physiological Ca 2+ signaling ex vivo and in vivo .