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Adenosine A2B receptor activation stimulates glucose uptake in the mouse forebrain

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Abstract

ATP consumption during intense neuronal activity leads to peaks of both extracellular adenosine levels and increased glucose uptake in the brain. Here, we investigated the hypothesis that the activation of the low-affinity adenosine receptor, the A2B receptor (A2BR), promotes glucose uptake in neurons and astrocytes, thereby linking brain activity with energy metabolism. To this end, we mapped the spatiotemporal accumulation of the fluorescent-labelled deoxyglucose, 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose (2-NBDG), in superfused acute hippocampal slices of C57Bl/6j mice. Bath application of the A2BR agonist BAY606583 (300 nM) triggered an immediate and stable (>10 min) increase of the velocity of 2-NBDG accumulation throughout hippocampal slices. This was abolished with the pretreatment with the selective A2BR antagonist, MRS1754 (200 nM), and was also absent in A2BR null-mutant mice. In mouse primary astrocytic or neuronal cultures, BAY606583 similarly increased 3H-deoxyglucose uptake in the following 20 min incubation period, which was again abolished by a pretreatment with MRS1754. Finally, incubation of hippocampal, frontocortical, or striatal slices of C57Bl/6j mice at 37 °C, with either MRS1754 (200 nM) or adenosine deaminase (3 U/mL) significantly reduced glucose uptake. Furthermore, A2BR blockade diminished newly synthesized glycogen content and at least in the striatum, increased lactate release. In conclusion, we report here that A2BR activation is associated with an instant and tonic increase of glucose transport into neurons and astrocytes in the mouse brain. These prompt further investigations to evaluate the clinical potential of this novel glucoregulator mechanism.

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Abbreviations

A2BR(s):

Adenosine A2B receptor(s)

2-NBDG:

2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose

A1R:

Adenosine A1 receptor

FELASA:

Federation for Laboratory Animal Science Associations

KO:

Knockout

HEPES:

N-(2-hydroxyethyl)piperazine-N′-(2-ethanesulfonic acid)

DMSO:

Dimethyl sulfoxide

DNAse:

Deoxyribonuclease

DMEM:

Dulbecco’s modified Eagle’s medium

HDG:

3H-2-deoxyglucose

CyB:

Cytochalasin B

GLUT1:

Glucose transporter type 1

SEM:

Standard error of the mean

ANOVA:

Analysis of variance

ADA:

Adenosine deaminase

DL-TBOA:

DL-threo-β-Benzyloxyaspartic acid

GABA:

γ-aminobutyric acid

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Acknowledgments

This study was financially supported by NARSAD, Santa Casa da Misercórdia, DARPA (09-68-ESR-FP-010), CAPES-FCT, and CNPq (Ciência sem Fronteiras) and co-funded by FEDER (QREN), through Programa Mais Centro under project CENTRO-07-ST24-FEDER-002006 and through Programa Operacional Factores de Competitividade—COMPETE and National funds (PTDC/SAU-OSM/105663/2008, EXPL/NEU-NMC/0671/2012 and Pest-C/SAU/LA0001/2013-2014) via FCT—Fundação para a Ciência e a Tecnologia.

Author Contributions

A.K., R.A.C., and D.R. conceived and designed the work. A.K., B.S.P., and C.L. carried out the experiments. R.O.B., J.M.M., and R.J.R. were responsible for the cell cultures. A.K. performed data analysis and wrote the first draft. A.K., C.L., R.A.C., and D.R. interpreted results and contributed to the writing and the edition of the manuscript. R.A.C., R.J.R., and D.R. supported the costs of the study. All authors have reviewed and commented on the manuscript.

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Authors and Affiliations

  1. CNC, Center for Neuroscience and Cell Biology of Coimbra, University of Coimbra, 3004-504, Coimbra, Portugal

    Cristina Lemos, Bárbara S. Pinheiro, Rui O. Beleza, Joana M. Marques, Ricardo J. Rodrigues, Rodrigo A. Cunha, Daniel Rial & Attila Köfalvi

  2. Institute for Interdisciplinary Research, University of Coimbra, Coimbra, Portugal

    Ricardo J. Rodrigues & Attila Köfalvi

  3. FMUC, Faculty of Medicine, University of Coimbra, Coimbra, Portugal

    Rodrigo A. Cunha

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  1. Cristina Lemos
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  2. Bárbara S. Pinheiro
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Correspondence to Attila Köfalvi.

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The authors have no conflict of interests to report and received no financial support or compensation from any individual or corporate entity over the past 3 years for research or professional services, and there are no personal holdings that could be perceived as constituting a potential conflict of interest.

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Lemos, C., Pinheiro, B.S., Beleza, R.O. et al. Adenosine A2B receptor activation stimulates glucose uptake in the mouse forebrain. Purinergic Signalling 11, 561–569 (2015). https://doi.org/10.1007/s11302-015-9474-3

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