Journal: EMBO Reports
Article Title: Cyst-independent oocyte phagocytosis builds the female reproductive reserve in mice
doi: 10.1038/s44319-025-00663-7
Figure Lengend Snippet: ( A ) The analysis of oocyte phagocytosis and development in cultured ovaries treated from c-17.5 to c-PD4 with autophagy inhibitors (3MA, MRT68921, BafA1, and CQ). Autophagy inhibition consistently reduced sacrifice and phagocytosis at c-PD1 and increased the survival of oocytes by c-PD4. Scale bar: 20 μm. ( B ) Statistical evaluations showed that suppression of autophagy significantly diminishes oocyte sacrifice and phagocytosis at c-PD1, demonstrating a sharp decline in the ratio of surviving oocytes with FLs and a decrease in the formation of ODs from sacrificed oocytes. The analysis included four ovaries per group, evaluating more than 70 oocytes per ovary. FL: control vs. 3-MA: p value = 5.9E-09; control vs. MRT: p value = 6.7E-08; control vs. BafA1: p value = 1.9E-08; control vs. CQ: p value = 1.1E-06; 3-MA vs. MRT: p value = 0.21; 3-MA vs. BafA1: p value = 0.79; 3-MA vs. CQ: p value = 0.003; MRT vs. BafA1: p value = 0.80; MRT vs. CQ: p value = 0.22; BafA1 vs. CQ: p value = 0.05. OD: control vs. 3-MA: p value = 2.3E-10; control vs. MRT: p value = 4.1E-10; control vs. BafA1: p value = 7.2E-09; control vs. CQ: p value = 7.8E-08; 3-MA vs. MRT: p value = 0.96; 3-MA vs. BafA1: p value = 0.54; 3-MA vs. CQ: p value = 0.66; MRT vs. BafA1: p value = 0.23; MRT vs. CQ: p value = 0.94; BafA1 vs. CQ: p value = 0.07. ( C , D ) Immunofluorescent staining of the ovarian morphology ( C ) and oocyte number counting ( D ) at c-PD4 showed an increased number of surviving oocytes in ovaries treated with autophagy inhibitors. Red - DDX4; Green - FOXL2; Blue - Hoechst. Scale bar: 100 μm. Data were from at least three ovaries per group. Data were presented as the mean ± SD. control vs. 3-MA: p value = 7.25E-05; control vs. MRT: p value = 0.034; control vs. BafA1: p value = 0.011; control vs. CQ: p value = 0.0054; 3-MA vs. MRT: p value = 0.012; 3-MA vs. BafA1: p value = 0.035; 3-MA vs. CQ: p value = 0.155; MRT vs. BafA1: p value = 0.96; MRT vs. CQ: p value = 0.71; BafA1 vs. CQ: p value = 0.95. Statistical significance was determined by ANOVA tests. P (a, b) < 0.05, P (a, c) < 0.05, P (b, c) < 0.05.
Article Snippet: In the autophagy inhibitors or Formin inhibitor treatment experiments, after 4 days of adherent culture, the Oct4-CreER T2 ;mTmG or Oct4-CreER T2 ;Rainbow ovaries at c-17.5 dpc were treated with or without autophagy signaling cascade inhibitors (5 mM 3-MA, M9281, Sigma-Aldrich; 400 nM MRT68921, S7949, Selleckchem; 0.2 nM Bafilomycin A1, HY-100558, MedChemExpress; 4 μM Chloroquine, HY-17589A, MedChemExpress) or Formin inhibitor (20 μM SMIFH2, HY-16931, MedChemExpress) for 3 or 7 days, which media was changed every other day.
Techniques: Cell Culture, Inhibition, Control, Staining
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