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A Simple Spectrofluorometric Assay to Measure Total Intracellular Magnesium by a Hydroxyquinoline Derivative

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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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Notes

  1. “Magnesium” or “Mg” refer to the total pool of magnesium, i.e. free plus bound forms; “Mg2+” refers specifically to the ionized or free form.

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Acknowledgments

This work was supported by an ex-60% MIUR grant from the University of Bologna to Stefano Iotti, by a PRIN 2004 grant to Stefano Iotti, by the FIRB LATEMAR grant (http://www.latemar.polito.it), and MIUR, Linea D1 grant 2005–2007 to Federica I. Wolf and by Fondazione Cassa di Risparmio di Bologna to Luca Prodi. Cytofluorimetric assays were performed at the Interdepartmental Center for Biotechnological Research of the University of Bologna. We are in debt with Micaela Fabbri for the AAS measurements.

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Authors and Affiliations

  1. Dipartimento di Biochimica “G. Moruzzi”, Università degli Studi di Bologna, Via S. Donato 19/2, 40127, Bologna, Italy

    Giovanna Farruggia & Giulia Andreani

  2. Dipartimento di Medicina Clinica e Biotecnologia Applicata “D. Campanacci”, Università degli Studi di Bologna, Via Massarenti, 9, 40138, Bologna, Italy

    Stefano Iotti

  3. Dipartimento di Chimica “G. Ciamician”, Università degli Studi di Bologna, via Selmi 2, 40126, Bologna, Italy

    Luca Prodi, Nelsi Zaccheroni & Marco Montalti

  4. Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT, 84602, USA

    Paul B. Savage

  5. Istituto di Patologia Generale e Centro di Ricerca sul Cancro “Giovanni XXIII”, Facoltà di Medicina, Università Cattolica del Sacro Cuore, Largo F. Vito 1, 00168, Rome, Italy

    Valentina Trapani & Federica I. Wolf

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  1. Giovanna Farruggia
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  2. Stefano Iotti
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  3. Luca Prodi
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  9. Federica I. Wolf
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Correspondence to Giovanna Farruggia, Stefano Iotti or Federica I. Wolf.

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Farruggia, G., Iotti, S., Prodi, L. et al. A Simple Spectrofluorometric Assay to Measure Total Intracellular Magnesium by a Hydroxyquinoline Derivative. J Fluoresc 19, 11–19 (2009). https://doi.org/10.1007/s10895-008-0374-6

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  • DOI: https://doi.org/10.1007/s10895-008-0374-6

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