The variation of the training period time and velocity was adjusted for each protocol and their specific sessions. Figure 1 Schematical figure depicting the treadmill exercise training protocol.

The time sessions, speed and duration depict the intensity of exercise training throughout the period in which exercise training protocol was performed. Exercise training protocol applied from 21- until 90-days-old (A); and applied from 21- until 50-days-old or from 60- until 90-days-old (B). Food intake After weaning, rats from all groups were weighed, and food intake was determined every week by non-ingested chow. Food intake was calculated for each animal as chow consumed divided by bw. The total area under the curve (AUC) of food consumption throughout experimental protocol was calculated. Intravenous glucose tolerance test (ivGTT) At 91-day-old, rats from all groups Protein Tyrosine Kinase inhibitor underwent a surgery for the silicone cannula implantation into the right jugular vein, as previously described [29]. At 24 h after the surgery, and after to be AICAR mw fasted overnight (12 h; 7:00 PM to 7:00 AM) the rats received a glucose infusion (1 g/kg bw) by a cannula implanted in the right jugular vein. Blood samples were collected in heparinized syringes at 0 (before glucose administration), 5, 15, 30 and 45 min after the glucose administration. Plasma samples were stored at -20°C PI3K inhibitor for

determination of glucose concentrations by the glucose oxidase method (Gold Aanlisa®; Belo Horizonte/MG, Brazil). The AUC of glycemia throughout the ivGTT was calculated. Autonomic nerves activity assessment At 91-day-old, a batch of rats from all of the experimental groups,

after to be fasted overnight was subsequently anesthetized with thiopental (45 mg/kg bw). As previously described [29], surgical longitudinal incisions were made on the anterior cervical region. Under the dissection microscope, the nerve bundle of the left superior branch of the upper vagus nerve was severed from the carotid artery close to the trachea. The nerve trunk was pulled with a fine Megestrol Acetate cotton line, and a pair of recording silver electrodes (0.6 mm diameter), similar to a hook, were placed under the nerve. The nerve was covered with silicone oil to prevent dehydration. The electrode was connected to an electronic device (Bio-Amplificator, Insight®; Riberão Preto/SP, Brazil), which amplified the electrical signals up to 10,000 times, and the low and high frequencies, 1–80 kHz, were filtered. The neural signal output was acquired by an Insight interface (Insight®; Riberão Preto/SP, Brazil), viewed online and stored by a personal computer running software developed by Insight (Bio-Amplificator, Insight®; Riberão Preto/SP, Brazil). During all data acquisition, the animals were placed in a Faraday cage to avoid any electromagnetic interference.