AMPK activators have compound and concentration-specific effects on brain metabolism.

AMP-activated protein kinase (AMPK) is a key sensor of energy balance playing important roles in the balancing of anabolic and catabolic activities. The high energy demands of the brain and its limited capacity to store energy indicate that AMPK may play a significant role in brain metabolism. Here, we activated AMPK in guinea pig cortical tissue slices, both directly with A769662 and PF 06409577, and indirectly, using AICAR and metformin. We studied the resultant metabolism of [1- 13 C]glucose and [1,2- 13 C]acetate using NMR spectroscopy. We found distinct activator concentration dependent effects on metabolism which ranged from decreased metabolic pool sizes at EC 50 activator concentrations with no expected stimulation in glycolytic flux, to increased aerobic glycolysis and decreased pyruvate metabolism with certain activators. Further, activation with direct vs indirect activators produced distinct metabolic outcomes at both low (EC 50 ) and higher (EC 50 x 10) concentrations. Specific direct activation of β1-containing AMPK isoforms with PF 06409577 resulted in increased Krebs cycle activity, restoring pyruvate metabolism while A769662 increased lactate and alanine production as well as labelling of citrate and glutamine. These results reveal a complex metabolic response to AMPK activators in brain beyond increased aerobic glycolysis and indicate that further research is warranted into their concentration and mechanism-dependent impact.