Metformin and 2DG activate AMPK in hippocampal CA1 neurons.

Abstract

<p>A) Schematic of the AMPK-mTOR pathway. B) AMPK is activated 30 min after exposure to 2DG (10 mM, p = 0.019, n = 9), metformin (5 µM, p = 0.005, n = 6), or phenformin (10 µM, p = 0.018, n = 6). Hippocampal slices were incubated in ACSF and drug for 30 minutes and subjected to western blot with anti-phospho-Thr172-AMPK antibody followed by βIII-tubulin as a loading control. Representative western blots of duplicate lanes are shown, together with their quantification from at least 10 samples per condition (C). D) ATP levels are reduced in the presence of 10 mM 2DG. Slices were incubated in ACSF+10 mM 2DG (n = 3) as above. Tissue was lysed and subjected to a CellTiter-Glo ATP assay (Promega). E) 2DG activates AMPK in cell bodies of the pyramidal layer (PL) and dendrites of the stratum radiatum (SR). Anti-phospho-Thr172-AMPK immunoreactivity is displayed in green. Neu-N is displayed in red. Scale bar: 10 µm.</p