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Diffusion of d-glucose measured in the cytosol of a single astrocyte

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Abstract

Astrocytes interact with neurons and endothelial cells and may mediate exchange of metabolites between capillaries and nerve terminals. In the present study, we investigated intracellular glucose diffusion in purified astrocytes after local glucose uptake. We used a fluorescence resonance energy transfer (FRET)-based nano sensor to monitor the time dependence of the intracellular glucose concentration at specific positions within the cell. We observed a delay in onset and kinetics in regions away from the glucose uptake compared with the region where we locally super-fused astrocytes with the d-glucose-rich solution. We propose a mathematical model of glucose diffusion in astrocytes. The analysis showed that after gradual uptake of glucose, the locally increased intracellular glucose concentration is rapidly spread throughout the cytosol with an apparent diffusion coefficient (D app) of (2.38 ± 0.41) × 10−10 m2 s−1 (at 22–24 °C). Considering that the diffusion coefficient of d-glucose in water is D = 6.7 × 10−10 m2 s−1 (at 24 °C), D app determined in astrocytes indicates that the cytosolic tortuosity, which hinders glucose molecules, is approximately three times higher than in aqueous solution. We conclude that the value of D app for glucose measured in purified rat astrocytes is consistent with the view that cytosolic diffusion may allow glucose and glucose metabolites to traverse from the endothelial cells at the blood–brain barrier to neurons and neighboring astrocytes.

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Acknowledgments

The authors thank Wolf B. Frommer for providing the plasmid FLII12PGLU-700μ∆6 (http://www.addgene.org). We thank Dr. Helena H. Chowdhury and Dr. Nina Vardjan for valuable help with plasmid multiplication. This work was supported by Grants #P3-310, #J3-4146 from the Slovenian Research Agency (ARRS) and COST (European Cooperation in Science and Technology) action BM1002. M.L. acknowledges the support of ARRS through Program P1-0201 and Project J1-4148. He also thanks Dr. Andrej Lajovic for discussions and help with numerical manipulation of the data.

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Authors and Affiliations

  1. LN-MCP, Faculty of Medicine, Institute of Pathophysiology, University of Ljubljana, Zaloška cesta 4, 1000, Ljubljana, Slovenia

    Marko Kreft, Mateja Prebil & Robert Zorec

  2. Department of Biology, Biotechnical Faculty, University of Ljubljana, Vecna pot 111, Ljubljana, Slovenia

    Marko Kreft

  3. Celica Biomedical Center, Tehnološki park 24, Ljubljana, Slovenia

    Marko Kreft & Robert Zorec

  4. Faculty of Chemistry and Chemical Technology, University of Ljubljana, Aškerčeva 5, 1000, Ljubljana, Slovenia

    Miha Lukšič

  5. Faculty of Pharmacy, University of Ljubljana, Aškerčeva 7, 1000, Ljubljana, Slovenia

    Tomaž M. Zorec

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  1. Marko Kreft
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  2. Miha Lukšič
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Correspondence to Marko Kreft.

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Kreft, M., Lukšič, M., Zorec, T.M. et al. Diffusion of d-glucose measured in the cytosol of a single astrocyte. Cell. Mol. Life Sci. 70, 1483–1492 (2013). https://doi.org/10.1007/s00018-012-1219-7

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