Obesity is associated to reduction of glucose transporter 4 (GLUT4) and/or impaired insulin signaling pathway in skeletal muscle(1). Muscle contraction culminates in 5′ AMP-activated protein kinase (AMPK) activation and GLUT4 translocation(2). Training stimulates peroxisome proliferator-activated receptor-γ coactivator (PGC1α) transcription in muscle which is involved in GLUT4 regulation(3). We aimed to evaluate the effects of continuous or intermittent training on AMPK (coded by Prkaa2), PGC1α (Ppargc1a gene) and GLUT4 (Slc2a4 gene) expression in muscle of diet-induced obese rats. Sixty male Wistar rats aged 90 days were divided: sedentary control (SC), continuous exercise control (CEC), intermittent exercise control (IEC), sedentary obese (SO), continuous exercise obese (CEO), intermittent exercise obese (IEO). All animals received standard chow and obese also received high fat diet during 16 weeks (wk). Insulin tolerance test and the double effort test were performed after 8 wk of diet and at the end of 8 wk of training using blood collected from the tail distal end. The CEC and CEO groups trained for 30 min at 90% of anaerobic threshold, 3x/wk, 8 wk. The IEC and IEO performed 11 efforts of 2 min, 1 min of interval, 3x/wk, 8 wk, over the delta zero to 120% of the critical load. Rats were anesthetized with Pentobarbital sodium 40mg/kg, ip, for removal of gastrocnemius muscle. RT-PCR and Western blotting were performed for mRNA and protein analysis. The experimental protocol was approved by the Ethics Committee for Animal Research (#74/2009). ANOVA was used for comparison among means with post-hoc (Tukey) if necessary, considering p<0.05. Obese presented similar gain of weight after 8 wk of diet (23%,p<0.05), which was softened after training (CEC-31%, EIC-28%,p<0.01vsSC; CEO-54%, IEO-44%,p<0.001vsSO).Obese presented reduced insulin sensitivity (SC=2.3+/-0.23; SO=1.8+/-0.2%/min,p<0.05), but both protocols reverted this condition (CEO=2.8+-0.7;IEO=2.4+-0.4%/min,p<0.05vsSO). Prkaa2 mRNA was increased with exercise (CEC=70%,EIC=72%,p<0.01vsSC;COE=75%, EIO=59%,p<0.05vsSO), AMPKα2 protein content was increased in exercised controls (CEC=47%,IEC=51%,p<0.05vsSC). pAMPKα2 was reduced in SO (39%,p<0.05vsSC), and increased after training (CEO=264%,IEO=299%,p<0.01vsSO). Both protocols were efficient to increase Ppargc1a mRNA expression (CEC=62%,IEC=126%,p<0.05vsSC; CEO=104%, IEO=152%,p<0.05vsSO). Continuous exercise provoked an increase (53%) in Slc2a4 expression in obese (p<0.05), Intermittent training improved control (55%,p<0.05). Eight weeks of both training protocols were able to soften the gain body mass, improve the insulin sensitivity and increase aerobic capacity, promoting increase of the protein expression involved in muscle metabolism.