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Assessment of AMPK-Stimulated Cellular Long-Chain Fatty Acid and Glucose Uptake

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AMPK

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1732))

Abstract

Here we describe an assay for simultaneous measurement of cellular uptake rates of long-chain fatty acids (LCFA) and glucose that can be applied to cells in suspension. The uptake assay includes the use of radiolabeled substrates at such concentrations and incubation periods that exact information is provided about unidirectional uptakes rates. Cellular uptake of both substrates is under regulation of AMPK. The underlying mechanism includes the translocation of LCFA and glucose transporters from intracellular membrane compartments to the cell surface, leading to an increase in substrate uptake. In this chapter, we explain the principles of the uptake assay before detailing the exact procedure. We also provide information of the specific LCFA and glucose transporters subject to AMPK-mediated subcellular translocation. Finally, we discuss the application of AMPK inhibitors and activators in combination with cellular substrate uptake assays.

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Authors and Affiliations

  1. Department of Molecular Genetics, CARIM School for Cardiovascular Diseases, Maastricht University, Maastricht, The Netherlands

    Joost J. F. P. Luiken, Jan F. C. Glatz, Will A. Coumans, Dipanjan Chanda & Miranda Nabben

  2. Department of Pathology, CARIM School for Cardiovascular Diseases, Maastricht University, Maastricht, The Netherlands

    Dietbert Neumann

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  1. Joost J. F. P. Luiken
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  2. Dietbert Neumann
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  3. Jan F. C. Glatz
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  4. Will A. Coumans
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  5. Dipanjan Chanda
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  6. Miranda Nabben
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Correspondence to Joost J. F. P. Luiken .

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Editors and Affiliations

  1. Department of Pathology CARIM School for Cardiovascular Diseases, Maastricht University, Maastricht, The Netherlands

    Dietbert Neumann

  2. Department EMD, Inserm, U1016, Institut Cochin, CNRS, UMR8104, Université Paris Descartes, Paris, France

    Benoit Viollet

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Luiken, J.J.F.P., Neumann, D., Glatz, J.F.C., Coumans, W.A., Chanda, D., Nabben, M. (2018). Assessment of AMPK-Stimulated Cellular Long-Chain Fatty Acid and Glucose Uptake. In: Neumann, D., Viollet, B. (eds) AMPK. Methods in Molecular Biology, vol 1732. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7598-3_22

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  • DOI: https://doi.org/10.1007/978-1-4939-7598-3_22

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