Concerning the signaling process of the hormone, it has been demonstrated that modulates the activation of AKT, protein involved in postnatal cardiac growth and coronary angiogenesis [19], time-dependently and is associated to PI3K and AMPK phosphorylation, protein kinases that have central role in the signaling processes for energy metabolism and cell growth [2], [3], [6] and [23]. In this paper we hypothesized that obesity and heart hypertrophy caused by early life overnutrition could GDC 0199 be related to changes in ghrelin signaling in heart, mainly in ghrelin-associated proteins (AKT, PI3K and AMPK), inducing a new pattern of heart growth or remodeling

and heart energetic availability. Virgin female Swiss mice were time crossed at 3 months of age. During pregnancy and lactation, they were singly housed in

individual cages and had ad libitum water and a standard pellet diet. After birth, litters were adjusted to nine pups per dam. At postnatal day 3, to induce early postnatal overnutrition, the number of pups per dam was adjusted to three male mice per litter to form the small litter (SL) [35], whereas litters containing nine pups per mother served as controls (normal litter (NL). To complete sample size only one male mice of each litter was used in order to discard pups of the same litter [48]. After weaning at postnatal day 21, mice were housed with three mice per cage with free access to water and standard chow in a temperature-controlled ifenprodil buy Seliciclib room with a 12 h light:12 h darkness cycle. They were weighed weekly and were killed at 180 days of age. Before the sacrifice mice were fasted overnight, injected with heparin (5000 U/kg), and then anesthetized with Avertin (0.3 g/kg body weight, via i.p. injection). They were cared for in accordance with the Animal Care and Use Committee of the Biology Institute of the State University of Rio de Janeiro, which based its analysis on the principles described

in the Guide for Care and Use of Laboratory Animals [5]. After a 12-h fast, blood glucose concentration was measured from blood droplets removed from the tail vein of 180 day old SL and age-matched NL mice with a glucometer (Accu-Chek, Roche, Sao Paulo, Brazil). Acylated ghrelin levels were determined in plasma using a commercial assay kit (Millipore, ELISA Kit, Rat/Mouse Ghrelin active). Blood sample was obtained under anesthesia by heart puncture, collected into a centrifuge tube containing K3 EDTA to achieve a final concentration of 1.735 mg/mL and treated with Pefabloc followed by immediate centrifugation (3000 rpm for 10 min at 4 °C). Plasma samples were acidified with HCl to a final concentration of 0.05 N and stored at −20 °C until assayed.