Dynamic imaging of cytosolic zinc in Arabidopsis roots combining FRET sensors and RootChip technology

* 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 Zn2+ homeostasis, the measureme...

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Hauptverfasser: Lanquar, Viviane (VerfasserIn) , Großmann, Guido (VerfasserIn)
Dokumenttyp: Article (Journal)
Sprache:Englisch
Veröffentlicht: 23 December 2013
In: The new phytologist
Year: 2014, Jahrgang: 202, Heft: 1, Pages: 198-208
ISSN:1469-8137
DOI:10.1111/nph.12652
Online-Zugang:Verlag, kostenfrei, Volltext: http://dx.doi.org/10.1111/nph.12652
Verlag, kostenfrei, Volltext: http://onlinelibrary.wiley.com/doi/10.1111/nph.12652/abstract
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Verfasserangaben:Viviane Lanquar, Guido Grossmann, Jan L. Vinkenborg, Maarten Merkx, Sébastien Thomine and Wolf B. Frommer

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520 |a * 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 Zn2+ 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 Zn2+ concentrations in Arabidopsis root cells. * Our experiments provide estimates of cytosolic free Zn2+ concentrations in Arabidopsis root cells grown under sufficient (0.4 nM) and excess (2 nM) Zn2+ supply. In addition, monitoring the dynamics of cytosolic [Zn2+] 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. 
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