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. 2022 Jun 15;8(24):eabl8070. doi: 10.1126/sciadv.abl8070

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Copyright © 2022 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).

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Fig. 1. — (A to D) Oocyte-specific deletion of Drp1. DRP1 (green) and DNA (pink) in Drp1^fl/fl oocytes (A) but Drp1^Δ/Δ oocytes lacked DRP1 (B). Dashed circles represent oocyte margins. (C) DRP1 fluorescence quantification. AU, arbitrary unit. (D) Western blots of DRP1. Each lane contains 50 GV-stage oocytes from 4- to 5-week-old mice. β-Actin was used as a loading control. The experiments were repeated three times, and a representative image is shown. Mitochondrial morphology in GV-stage Drp1^fl/fl (E and F) and Drp1^Δ/Δ (G and H) oocytes. Branch length mean (I) and network branches mean (J) between Drp1^fl/fl and Drp1^Δ/Δ oocytes. Oocytes were labeled with mitotracker orange. Number of oocytes analyzed are shown (C, I, and J). (K) Increased mitochondrial ROS (mtROS) levels in Drp1^Δ/Δ oocytes. Numbers represent total number of oocytes. (L) Reduced MMP in Drp1^Δ/Δ oocytes compared to Drp1^fl/fl oocytes. Carbonyl cyanide-4-phenylhydrazone (FCCP) was used to demonstrate assay specificity. Numbers represent total number of oocytes. (M) Similar mitochondrial mass between Drp1^fl/fl and Drp1^Δ/Δ oocytes. MitoTracker Green signals in (L) were analyzed to calculate mitochondrial mass. ns, not significant. (N) Comparable mtDNA copy number between Drp1^fl/fl and Drp1^Δ/Δ oocytes. Numbers represent total number of oocytes. (O) Similar ATP levels between Drp1^Δ/Δ and Drp1^fl/fl oocytes. Numbers represent experimental replicates. Drp1^Δ/Δ oocytes have increased levels of FAD⁺⁺ (P) and NAD(P)H (Q) compared to Drp1^fl/fl controls. Total oocyte numbers shown. Representative autofluorescent images are also shown. (R) Comparable PBE rates in vitro. Numbers represent experimental replicates. (S to U) MII oocytes showing spindle (green) and DNA (blue). Chromosome spreads were measured as exemplified in (T) and (U). Total oocyte numbers shown in (S). Results show means ± SD and P values determined by two-tailed Student’s t test (C and I to S). F.C., fold change.