A Simple Spectrofluorometric Assay to Measure Total Intracellular Magnesium by a Hydroxyquinoline Derivative |
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Authors: | Giovanna Farruggia Stefano Iotti Luca Prodi Nelsi Zaccheroni Marco Montalti Paul B Savage Giulia Andreani Valentina Trapani Federica I Wolf |
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Institution: | Dipartimento di Biochimica G. Moruzzi, Università degli Studi di Bologna, Via S. Donato 19/2, 40127, Bologna, Italy. Giovanna.Farruggia@unibo.it |
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Abstract: | The intracellular behaviour of diaza-18-crown-6 appended with two H-substituted hydroxyquinoline groups (DCHQ1) was investigated
to explore its application as a new sensor for the evaluation of cell magnesium content and distribution. We used five cells
lines characterised by different contents of magnesium and different intracellular membrane-defined compartments. The main
result is the definition of the appropriate experimental conditions to quantitatively assess the total cell magnesium by fluorescence
spectroscopy. We showed that disrupting cells by sonication, DCHQ1 was capable to assess total cell magnesium in all cell
types examined, obtaining overlapping results with atomic absorption spectroscopy (AAS). This new analytical approach requires
very small cell samples and a simple fluorimetric technique, and can be a valid alternative to AAS. The fluorescent properties
of DCHQ1 in living cells are: (a) it consistently stains live cells, (b) it discriminates small variations of cell Mg contents,
(c) cell staining is stable for at least 30 min. We also investigated the role of lipophilic environment on DCHQ1 fluorescence
by mimicking cell membranes and described how the composition and structure of lipid vesicles affect Mg-DCHQ1 fluorescence.
Thus, DCHQ1 may offer important information also on magnesium distribution in living cells, providing a novel strategy to
map the intracellular compartmentalization of this cation.
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Keywords: | Magnesium Fluorescent probes Hydroxyquinoline derivatives Fluorescence spectroscopy Flow cytometry |
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