title: Dynamic imaging of cytosolic zinc in Arabidopsis roots combining FRET sensors and RootChip technology.
abstract: Zinc plays a central role in all living cells as a cofactor for enzymes and as a structural element enabling the adequate folding of proteins. In eukaryotic cells, metals are highly compartmentalized and chelated. Although essential to characterize the mechanisms of Zn(2+) homeostasis, the measurement of free metal concentrations in living cells has proved challenging and the dynamics are difficult to determine. Our work combines the use of genetically encoded Förster resonance energy transfer (FRET) sensors and a novel microfluidic technology, the RootChip, to monitor the dynamics of cytosolic Zn(2+) concentrations in Arabidopsis root cells. Our experiments provide estimates of cytosolic free Zn(2+) concentrations in Arabidopsis root cells grown under sufficient (0.4 nM) and excess (2 nM) Zn(2+) supply. In addition, monitoring the dynamics of cytosolic [Zn(2+) ] in response to external supply suggests the involvement of high- and low-affinity uptake systems as well as release from internal stores. In this study, we demonstrate that the combination of genetically encoded FRET sensors and microfluidics provides an attractive tool to monitor the dynamics of cellular metal ion concentrations over a wide concentration range in root cells.
authors.affiliation: Department of Plant Biology, Carnegie Institution for Science, 260 Panama St, Stanford, CA, 94305, USA.|CNRS, Institut des Sciences du Végétal, Saclay Plant Sciences, Avenue de la Terrasse, 91198, Gif-sur-Yvette, France.
authors.forename: Viviane
authors.initials: V
authors.lastname: Lanquar
authors.affiliation: Department of Plant Biology, Carnegie Institution for Science, 260 Panama St, Stanford, CA, 94305, USA.
authors.forename: Guido
authors.initials: G
authors.lastname: Grossmann
authors.affiliation: Laboratory of Chemical Biology, Eindhoven University of Technology, PO Box 513, 5600 MB, Eindhoven, the Netherlands.
authors.forename: Jan L
authors.initials: JL
authors.lastname: Vinkenborg
authors.affiliation: Laboratory of Chemical Biology, Eindhoven University of Technology, PO Box 513, 5600 MB, Eindhoven, the Netherlands.
authors.forename: Maarten
authors.initials: M
authors.lastname: Merkx
authors.affiliation: CNRS, Institut des Sciences du Végétal, Saclay Plant Sciences, Avenue de la Terrasse, 91198, Gif-sur-Yvette, France.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Thomine
authors.affiliation: Department of Plant Biology, Carnegie Institution for Science, 260 Panama St, Stanford, CA, 94305, USA.
authors.forename: Wolf B
authors.initials: WB
authors.lastname: Frommer
mesh_terms: D017360:Arabidopsis / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism*; D005110:Extracellular Space / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D042541:Intracellular Space / Q000378:metabolism; D010477:Perfusion; D018517:Plant Roots / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D015032:Zinc / Q000378:metabolism*
chemical_list: D015032:Zinc
keywords: Arabidopsis; nanosensor; nutrition; transport; zinc