Figure 1.

(A–E) Effect of cryopreservation and/or transplantation on oxygen consumption in homogenates of hemi-ovaries. Oxygen flux was evaluated using a substrate-uncoupler-inhibitor titration protocol (SUIT). Average O₂ consumption rates (OCR) measured at 5 points of the evaluation, namely (A) Basal (O₂ consumed due to oxidation of endogenous substrates); (B) LEAK (O₂ consumed due to oxidation of exogenous succinate); (C) OXPHOS (OCR related to oxidative phosphorylation); (D) Oligo (OCR not dependent on ATP synthesis); (E) ETS (maximal respiratory rate resulting from uncoupled mitochondrial respiration). Assessment of reserve mitochondrial capacity (Spare capacity) (F) and oxygen consumption directed to ATP synthesis (ATP-linked) (G) were also evaluated. (H) Representative oxygraph trace of hemi-ovary oxygen consumption during the substrate-uncoupler-inhibitor titration protocol (SUIT) (arrowheads). Respiration rates were measured at 37 °C. O₂ consumption was measured after sequential additions of 10 mM succinate (Succ) and 0.5 µM rotenone (Rot), 500 µM adenosine diphosphate (ADP), 0.1 µg/ml oligomycin (Oligo), Carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP) (0.1 to 0.4 µM) and 1 µM antimycin (AA). All results were expressed as the mean oxygen consumption rates in pmol of oxygen consumed per second/mg determined by mass of tissue. Data are expressed as the mean ± SEM. *p < 0.05 compared with Fresh non-transplanted group (Two-way ANOVA followed by Tukey’s post-hoc test).