Development of Fluorescent Glucose Bioprobes and Their Application on Real-Time and Quantitative Monitoring of Glucose Uptake in Living Cells

Authors

  • Hyang Yeon Lee,

    1. Department of Chemistry, Seoul National University, Seoul 151-747 (Korea), Fax: (+82) 28844025
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  • Jae Jeong Lee,

    1. Department of Chemistry, Seoul National University, Seoul 151-747 (Korea), Fax: (+82) 28844025
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  • Jongmin Park,

    1. Department of Chemistry, Seoul National University, Seoul 151-747 (Korea), Fax: (+82) 28844025
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  • Prof. Seung Bum Park

    Corresponding author
    1. Department of Chemistry, Seoul National University, Seoul 151-747 (Korea), Fax: (+82) 28844025
    2. Department of Biophysics and Chemical Biology, Seoul National University, Seoul 151-747 (Korea)
    • Department of Chemistry, Seoul National University, Seoul 151-747 (Korea), Fax: (+82) 28844025
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Abstract

We developed a novel fluorescent glucose bioprobe, GB2-Cy3, for the real-time and quantitative monitoring of glucose uptake in living cells. We synthesized a series of fluorescent glucose analogues by adding Cy3 fluorophores to the α-anomeric position of D-glucose through various linkers. Systematic and quantitative analysis of these Cy3-labeled glucose analogues revealed that GB2-Cy3 was the ideal fluorescent glucose bioprobe. The cellular uptake of this probe competed with the cellular uptake of D-glucose in the media and was mediated by a glucose-specific transport system, and not by passive diffusion. Flow cytometry and fluorescence microscopy analyses revealed that GB2-Cy3 is ten times more sensitive than 2-NBDG, a leading fluorescent glucose bioprobe. GB2-Cy3 can also be utilized for the quantitative flow cytometry monitoring of glucose uptake in metabolically active C2C12 myocytes under various treatment conditions. As opposed to a glucose uptake assay performed by using radioisotope-labeled deoxy-D-glucose and a scintillation counter, GB2-Cy3 allows the real-time monitoring of glucose uptake in living cells under various experimental conditions by using fluorescence microscopy or confocal laser scanning microscopy (CLSM). Therefore, we believe that GB2-Cy3 can be utilized in high-content screening (HCS) for the discovery of novel therapeutic agents and for making significant advances in biomedical studies and diagnosis of various diseases, especially metabolic diseases.

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