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Microscopic Visualization of Metabotropic Glutamate Receptors on the Surface of Living Cells Using Bifunctional Magnetic Resonance Imaging Probes

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    Research Article

    Microscopic Visualization of Metabotropic Glutamate Receptors on the Surface of Living Cells Using Bifunctional Magnetic Resonance Imaging Probes
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    † †Department of Chemistry and ‡School of Biological and Biomedical Sciences, Durham University, South Road, Durham DH1 3LE, England
    § High Field MR Centre, Max Planck Institute for Biological Cybernetics, Spemannstrasse 41, Tuebingen 72076, Germany
    *Fax: +49-8931872017. E-mail: [email protected] (A.M.).
    *E-mail: [email protected] (D.P.).
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    ACS Chemical Neuroscience

    Cite this: ACS Chem. Neurosci. 2014, 5, 2, 128–137
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    https://doi.org/10.1021/cn400175m
    Published November 19, 2013
    Copyright © 2013 American Chemical Society
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    Abstract

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    A series of bimodal metabotropic glutamate-receptor targeted MRI contrast agents has been developed and evaluated, based on established competitive metabotropic Glu receptor subtype 5 (mGluR5) antagonists. In order to directly visualize mGluR5 binding of these agents on the surface of live astrocytes, variations in the core structure were made. A set of gadolinium conjugates containing either a cyanine dye or a fluorescein moiety was accordingly prepared, to allow visualization by optical microscopy in cellulo. In each case, surface receptor binding was compromised and cell internalization observed. Another approach, examining the location of a terbium analogue via sensitized emission, also exhibited nonspecific cell uptake in neuronal cell line models. Finally, biotin derivatives of two lead compounds were prepared, and the specificity of binding to the mGluR5 cell surface receptors was demonstrated with the aid of their fluorescently labeled avidin conjugates, using both total internal reflection fluorescence (TIRF) and confocal microscopy.

    Copyright © 2013 American Chemical Society

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    Supporting Information

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    Synthesis of intermediate amines A and B, experimental methods, Figures S1–S7, and Videos S1 and S2. This material is available free of charge via the Internet at http://pubs.acs.org.

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    ACS Chemical Neuroscience

    Cite this: ACS Chem. Neurosci. 2014, 5, 2, 128–137
    Click to copy citationCitation copied!
    https://doi.org/10.1021/cn400175m
    Published November 19, 2013
    Copyright © 2013 American Chemical Society
    Request reuse permissions

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