title: Measuring intranuclear and nuclear envelope [Ca(2+)] vs. cytosolic [Ca (2+)].
abstract: Nuclear Ca(2+) regulates key cellular processes, including gene expression, apoptosis, assembly of the nuclear envelope, and nucleocytoplasmic transport. Quantification of subcellularly resolved Ca(2+) signals is, therefore, essential for understanding physiological and pathological processes in various cell types. However, the properties of commonly used Ca(2+)-fluorescent indicators in intracellular compartments may differ, thus affecting the translation of Ca(2+)-dependent fluorescence changes into quantitative changes of Ca(2+) concentration. Here, we describe technical approaches for reliable subcellular quantification of [Ca(2+)] in the cytoplasm vs. the nucleus and the nuclear envelope by in situ calibration of fluorescein-derived fluorescent indicators Fluo-4 and Fluo-5N.
authors.affiliation: Department of Pharmacology, University of California Davis, Davis, CA, 95616-8636, USA.
authors.forename: Senka
authors.initials: S
authors.lastname: Ljubojević
authors.forename: Donald M
authors.initials: DM
authors.lastname: Bers
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000737:chemistry; D051379:Mice; D057054:Molecular Imaging / Q000379:methods*; D009685:Nuclear Envelope / Q000378:metabolism*
chemical_list: D005456:Fluorescent Dyes; D002118:Calcium