title: Highlighted Ca²⁺ imaging with a genetically encoded 'caged' indicator.
abstract: Genetically encoded fluorescent indicators for bioimaging are powerful tools for visualizing biological phenomena in specified cell types or cellular compartments. However, available gene promoters or localization sequences are not applicable for visualizing all expression events. Furthermore, a visualization technique focusing on single cells or cellular compartments is required for characterizing specific cellular properties including individuality of cells in the cell population. To address these limitations, we developed a genetically encoded caged Ca(2+) indicator for which expression timing and location could be controlled. This indicator, PA-TNXL, comprises a Ca(2+)-binding protein and troponin between a photoactivatable FRET donor (PA-GFP) and a FRET quencher (dim variant of YFP). Ultraviolet irradiation activates the FRET Ca(2+) indicator. Using this indicator, we successfully imaged Ca(2+) dynamics in a given set of HeLa cells and cultured hippocampal neurons. This technology can be applied for developing other photoactivatable indicators, thereby opening a new area of biological research.
authors.affiliation: The Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki, Osaka 567-0047, Japan.
authors.forename: Tomoki
authors.initials: T
authors.lastname: Matsuda
authors.forename: Kazuki
authors.initials: K
authors.lastname: Horikawa
authors.forename: Kenta
authors.initials: K
authors.lastname: Saito
authors.forename: Takeharu
authors.initials: T
authors.lastname: Nagai
mesh_terms: D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D031541:Fluorescence Resonance Energy Transfer; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014336:Troponin / Q000378:metabolism
chemical_list: D011993:Recombinant Fusion Proteins; D014336:Troponin; D049452:Green Fluorescent Proteins; D002118:Calcium