is a key regulator of fat and carbohydrate metabolism. It has been postulated that defects in AMPK signaling could be responsible for some of the metabolic abnormalities of type 2 diabetes. In this study, we examined whether insulin-resistant obese Zucker rats have abnormalities in the AMPK pathway. We compared AMPK and ACC phosphorylation and the protein content of the upstream AMPK kinase LKB1 and the AMPKregulated transcriptional coactivator PPAR␥ coactivator-1 (PGC-1) in gastrocnemius of sedentary obese Zucker rats and sedentary lean Zucker rats. We also examined whether 7 wk of exercise training on a treadmill reversed abnormalities in the AMPK pathway in obese Zucker rats. In the obese rats, AMPK phosphorylation was reduced by 45% compared with lean rats. Protein expression of the AMPK kinase LKB1 was also reduced in the muscle from obese rats by 43%. In obese rats, phosphorylation of ACC and protein expression of PGC-1␣, two AMPK-regulated proteins, tended to be reduced by 50 (P ϭ 0.07) and 35% (P ϭ 0.1), respectively. There were no differences in AMPK␣1, -␣2, -1, -2, and -␥3 protein content between lean and obese rats. Training caused a 1.5-fold increase in AMPK␣1 protein content in the obese rats, although there was no effect of training on AMPK phosphorylation and the other AMPK isoforms. Furthermore, training also significantly increased LKB1 and PGC-1␣ protein content 2.8-and 2.5-fold, respectively, in the obese rats. LKB1 protein strongly correlated with hexokinase II activity (r ϭ 0.75, P ϭ 0.001), citrate synthase activity (r ϭ 0.54, P ϭ 0.02), and PGC-1␣ protein content (r ϭ 0.81, P Ͻ 0.001). In summary, obese insulin-resistant rodents have abnormalities in the LKB1-AMPK-PGC-1 pathway in muscle, and these abnormalities can be restored by training.acetyl-CoA carboxylase; peroxisome proliferator-activated receptor-␥ coactivator-1␣; exercise AMP-ACTIVATED PROTEIN KINASE (AMPK) is an energy-sensing enzyme that is activated by increases in AMP-to-ATP ratios through mechanisms involving allosteric modification and phosphorylation by one or more upstream AMPK kinases (AMPKKs) (11,25). AMPK is a heterotrimer that is formed by a catalytic ␣-subunit and the regulatory -and ␥-subunits. There are two isoforms of the ␣-and -subunits and three isoforms of the ␥-subunit (11, 25). Upon activation, AMPK switches on catabolic pathways that generate ATP, such as glucose transport and fatty acid oxidation, coupled with the inhibition of ATP-consuming (anabolic) pathways that include triglyceride and cholesterol synthesis. It has become apparent in recent years that AMPK plays a key role in mediating the acute and long-term effects of exercise on carbohydrate and fat metabolism in skeletal muscle. AMPK mediates, at least in part, exercise-stimulated increases in muscle glucose transport (3,15,34,35) and fatty acid oxidation (53, 57). The stimulatory effect of AMPK on fatty acid oxidation results from the phosphorylation and inhibition of acetyl-CoA carboxylase (ACC) by AMPK (10, 53, 59). There is evidence that AM...