Leptin decreases apoptosis and promotes the activation of primordial follicles through the phosphatidylinositol-3-kinase/protein kinase B pathway in cultured ovine ovarian tissue.

This study evaluated the effects of leptin on primordial follicle survival and activation after in vitro culture of ovine ovarian tissue and if leptin acts through the phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt) pathway. Ovarian fragments were fixed for histology (fresh control) or cultured for 7 days in control medium (α-MEM+) alone or supplemented with leptin (1, 5, 10, 25 or 50 ng/ml). Follicle morphology, activation and apoptosis were analyzed. Next, the fragments were cultured in the medium that showed the best results in the absence or the presence of the PI3K inhibitor (LY294002), and immunohistostaining of p-Akt protein was assessed. After culture, the percentage of normal follicles decreased (P < 0.05) in all treatments compared with the fresh control. Moreover, control medium and 1 ng/ml leptin had similar (P > 0.05) percentages of normal follicles, which were significantly higher than those in other treatments. However, culture with 1 ng/ml leptin maintained apoptosis similarly (P > 0.05) to that of the fresh control and lower (P < 0.05) than that in α-MEM+. Leptin did not influence follicle activation (P > 0.05) compared with the control medium (α-MEM+). Culture in 1 ng/ml leptin with LY294002 decreased the normal follicles and increased apoptosis, inhibited follicle activation (P < 0.05), and reduced p-Akt immunostaining, compared with the medium containing 1 ng/ml leptin without PI3K inhibitor. In conclusion, leptin at 1 ng/ml reduces apoptosis and promotes the activation of primordial follicles compared with the fresh control after in vitro culture of ovine ovarian tissue possibly through the PI3K/Akt pathway.

Effect of pre-exercise carbohydrate availability on fat oxidation and energy expenditure after a high-intensity exercise.

The aim of this study was to test the hypothesis that reduced pre-exercise carbohydrate (CHO) availability potentiates fat oxidation after an exhaustive high-intensity exercise bout. Eight physically active men underwent a high-intensity exercise (∼95% V̇O2max) until exhaustion under low or high pre-exercise CHO availability. The protocol to manipulate pre-exercise CHO availability consisted of a 90-min cycling bout at ∼70% V̇O2max + 6 × 1-min at 125% V̇O2max with 1-min rest, followed by 48 h under a low- (10% CHO, low-CHO availability) or high-CHO diet (80% CHO, high-CHO availability). Time to exhaustion was shorter and energy expenditure (EE) lower during the high-intensity exercise in low- compared to high-CHO availability (8.6±0.8 and 11.4±1.6 min, and 499±209 and 677±343 kJ, respectively, P<0.05). Post-exercise EE was similar between low- and high-CHO availability (425±147 and 348±54 kJ, respectively, P>0.05), but post-exercise fat oxidation was significantly higher (P<0.05) in low- (7,830±1,864 mg) than in high-CHO availability (6,264±1,763 mg). The total EE (i.e., exercise EE plus post-exercise EE) was similar between low- and high-CHO availability (924±264 and 1,026±340 kJ, respectively, P>0.05). These results suggest that a single bout of high-intensity exercise performed under low-CHO availability increased post-exercise fat oxidation, and even with shorter exercise duration, both post-exercise EE and total EE were not impaired.