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This study was designed to examine potential in vivo mechanisms of AMP-activated protein kinase (AMPK) phosphorylation inhibition and its downstream signaling consequences during the recovery period after a single bout of sprint exercise. Sprint exercise induces Thr¹⁷²-AMPK phosphorylation and increased PGC-1α mRNA, by an unknown mechanism. Muscle biopsies were obtained in 15 young healthy men in response to a 30-s sprint exercise (Wingate test) randomly distributed into two groups: the fasting (n = 7, C) and the glucose group (n = 8, G), who ingested 75 g of glucose 1 h before exercising to inhibit AMPKα phosphorylation. Exercise elicited different patterns of Ser²²¹-ACCβ, Ser⁴⁷³-Akt and Thr⁶⁴²-AS160 phosphorylation, during the recovery period after glucose ingestion. Thirty minutes after the control sprint, Ser⁴⁸⁵-AMPKα1/Ser⁴⁹¹-AMPKα2 phosphorylation was reduced by 33 …