title: Measuring Ca2+inside intracellular organelles with luminescent and fluorescent aequorin-based sensors.
abstract: GFP-Aequorin Protein (GAP) can be used to measure [Ca2+] inside intracellular organelles, both by luminescence and by fluorescence. The low-affinity variant GAP3 is adequate for ratiometric imaging in the endoplasmic reticulum and Golgi apparatus, and it can be combined with conventional synthetic indicators for simultaneous measurements of cytosolic Ca2+. GAP is bioorthogonal as it does not have mammalian homologues, and it is robust and functionally expressed in transgenic flies and mice, where it can be used for Ca2+measurements ex vivo and in vivo to explore animal models of health and disease. This article is part of a Special Issue entitled: ECS Meeting edited by Claus Heizmann, Joachim Krebs and Jacques Haiech.
authors.affiliation: 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. Electronic address: talonso@ibgm.uva.es.
authors.forename: María Teresa
authors.initials: MT
authors.lastname: Alonso
authors.affiliation: 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.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Rojo-Ruiz
authors.affiliation: 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.
authors.forename: Paloma
authors.initials: P
authors.lastname: Navas-Navarro
authors.affiliation: 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.
authors.forename: Macarena
authors.initials: M
authors.lastname: Rodríguez-Prados
authors.affiliation: 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. Electronic address: jgsancho@ibgm.uva.es.
authors.forename: Javier
authors.initials: J
authors.lastname: García-Sancho
mesh_terms: D000331:Aequorin / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D004175:Diptera; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D049449:Luminescence; D051379:Mice; D008822:Mice, Transgenic; D015388:Organelles / Q000378:metabolism*
chemical_list: D049452:Green Fluorescent Proteins; D000331:Aequorin; D002118:Calcium
keywords: Aequorin; Biosensor; Calcium; Endoplasmic reticulum; GECI; GFP; Golgi apparatus; Organelles