dependent on glucose metabolism. We invite the reader to
perform a similar test to verify the glycolytic profile of the
cells to be tested.- Figure5 shows an example of mitochondrial respiration test on
primary neuronal culture (Fig. 5a) in presence (Glc), or
absence (Ctrl), of glucose as metabolic substrate. The results
proposed here show that in absence of glucose, the OCR values
of basal respiration and maximal respiratory capacity are abro-
gated (Fig.5b and c), confirming that OCR values are depen-
dent on glucose metabolism. We invite the reader to perform a
similar test to verify the respiratory profile of the cells to be
tested. - Using Seahorse XFe technology it is possible to monitor cell
metabolism in presence of different energy substrates, includ-
ing lactate (10 mM Lac) (Fig.5d–f) and the ketone bodies 3-β
hydroxybutyrate (βHBA) and acetoacetate (AcAc) (Fig.5g–l).
Results show that injection of ketone bodies induces a readily
increase of OCR, suggesting that ketone bodies are metabo-
lized by neuronal mitochondria.
Acknowledgments
This work was supported by the French Foundation pour la coop-
eration Scientifique—Plan Alzheimer 2008–2012 (Senior Innova-
tive Grant 2013 to V.V.) and by the Fondation Vaincre Alzheimer,
nFR-16071p (to V.V.) and in part through the Labex DISTALZ
(Development of Innovative Strategies for a Transdisciplinary
Approach to Alzheimer’s Disease). This work received a financial
support from INSERM, UNIVERSITE DE LILLE II, a special
financial support from the Association pour l’Etude des Anomalies
Conge ́nitales Neurodev of Pr. B. Poupard (to J.K. and P.M.) and
the support of G. Mulliez.References
- Hardie DG (2007) AMP-activated/SNF1 pro-
tein kinases: conserved guardians of cellular
energy. Nat Rev Mol Cell Biol 8
(10):774–785. https://doi.org/10.1038/
nrm2249 - Hardie DG (2011) AMP-activated protein
kinase: an energy sensor that regulates all
aspects of cell function. Genes Dev 25
(18):1895–1908. https://doi.org/10.1101/
gad.17420111 - Viollet B, Horman S, Leclerc J, Lantier L,
Foretz M, Billaud M, Giri S, Andreelli F
(2010) AMPK inhibition in health and disease.
Crit Rev Biochem Mol Biol 45(4):276–295.
https://doi.org/10.3109/10409238.2010.
488215- Barnes K, Ingram JC, Porras OH, Barros LF,
Hudson ER, Fryer LG, Foufelle F, Carling D,
Hardie DG, Baldwin SA (2002) Activation of
GLUT1 by metabolic and osmotic stress:
potential involvement of AMP-activated pro-
tein kinase (AMPK). J Cell Sci 115
(Pt 11):2433–2442 - Abbud W, Habinowski S, Zhang JZ,
Kendrew J, Elkairi FS, Kemp BE, Witters LA,
Ismail-Beigi F (2000) Stimulation of
Seahorse Analysis of AMPK-Regulated Metabolic Fluxes 303