Figure 2
Mitochondrial dynamics in cumulus cells for different IVM periods (0, 4, 24, and 48 h). (A) Representative images of TMRM-stained cumulus cell mitochondria observed live by confocal microscopy at 63X after different IVM periods (0, 4, 24, and 48 h). (B) Manual quantification of mitochondrial network phenotypes reported as a percentage in each cumulus cell from 10 COCs for different IVM periods (0, 4, 24, and 48 h). Mitochondria were manually classified as very long, long, intermediate, and short, depending on their length in each cumulus cell. (C) Quantification of mitochondrial length distribution with the Momito algorithm for each IVM period. (D) EMD quantification of the differences between mitochondrial length distribution for different IVM periods. For each condition, EMD values were first calculated between each cell within a distribution type, and the average for that distribution to calculate the experimental variation for each IVM period presented as black histograms. Striped histograms refer to the EMD between distribution types (0 h Vs. 4 h, 4 h Vs. 24 h, 24 h Vs. 48 h). Each cell within a distribution type was compared to the average for each of the other distributions. (E) Mitochondrial aspect ratio and (F) Mitochondrial connectivity changes during IVM. All data are expressed as the average of three independent experiments ± SEM (10 COCs per replicate for each IVM time). (* p < 0.05; *** p < 0.001). Different letters indicate statistically significant differences (p < 0.05).
