Figure 3
Effects of FSH on mitochondrial dynamics and activity in cumulus cells, as well as their metabolic status during IVM. (A) Representative images of TMRM-stained mitochondria in cumulus cells exposed to FSH or not (Ct) for 2 h and 24 h of IVM. (B) Momito analysis of mitochondrial length distribution for each IVM condition. (C) The calculated EMD values for each culture condition (black histograms represent experimental variability for each IVM time; striped histograms refer to the measured statistical difference between two length distributions). (D) Momito analysis of mitochondrial connectivity for each IVM condition. Data of TMRM staining are expressed as the average of three independent experiments ± SEM (10 COCs for each IVM time). (E) Real-time kinetics of oxygen consumption rates (OCR) measured in COC during IVM by seahorse analyzer in response to FSH relative to control, which was set at 100% at 0 h. (F) Changes in the relative basal respiration measured by seahorse analyzer in control COCs and COCs exposed to FSH for different IVM periods. (G) Real-time kinetics of relative extracellular acidification rates (ECAR) measured in COCs by seahorse analyzer during IVM after FSH injection compared to control, which was set at 100% at 0 h. (H) Changes in basal glycolysis percentage measured by seahorse analyzer in control COCs and COCs exposed to FSH for different IVM periods. All data of seahorse analysis are expressed as the average of four independent experiments ± SEM (20 COCs by well in triplicate). (I) Lactate production in COCs cultured 2 h with FSH compared to control. Data of lactate measurements are expressed as the average of three independent experiments ± SEM with 20 COCs by well in triplicate per replicate, and different letters indicate statistically significant differences (p < 0.05). (* p < 0.05; *** p < 0.001).
