Resveratrol stimulates AMP kinase activity in neurons

Dasgupta and Milbrandt. 10.1073/pnas.0610068104.

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Fig. 7. Resveratrol activates AMP kinase in cortical neurons through LKB1 and calcium/calmodulin-dependent protein kinase b(CaMKKb). (A) Cortical neuron cultures were established from embryonic day (E)13.5 Lkb1^flox/flox embryos and infected with lentivirus-expressing Cre recombinase (FCIV-Cre) or GFP only (FUGW control). Lentiviral infection of cortical neurons was monitored by GFP fluorescence (A). A Western blot using LKB1 antibody demonstrated complete loss of Lkb1 in Cre-expressing cortical neurons (B). Lkb1^flox/flox cortical neurons were infected with FUGW or FCIV-Cre and treated as indicated. AMP kinase (AMPK) or acetyl-CoA carboxylase (ACC) was immunoprecipitated from neuronal lysates, and Western blots were probed with the respective phospho-specific antibodies. Resveratrol-mediated AMPK (C) and ACC phosphorylation (D) was significantly reduced upon Lkb1 excision by FCIV-Cre and in neurons treated with 2.5 mM CaMKKb inhibitor STO 609. No inhibition of resveratrol-stimulated AMPK or ACC phosphorylation was observed in neurons treated with SIRT1 inhibitors. Lysates from 5-aminoimidazole-4-carboxamide-1-b-D-ribofuranoside (AICAR)-treated cortical neurons are included as a positive control. (E and F) Embryonic cortical neurons were cultured from wild-type and SIRT1-deficient littermates derived from SIRT1 heterozygous matings. Western blot analysis with phospho-specific antibodies revealed that resveratrol stimulated AMPK (E) and ACC phosphorylation (F) equivalently in wild-type and SIRT1-deficient cortical neurons. Levels of total AMPK and ACC are shown in the C--F (Lower). Resv, resveratrol.

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