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. 2023 Jun 18;22(9):e13904. doi: 10.1111/acel.13904

FIGURE 5.

FIGURE 5

Deletion of Ido1 or Qprt led to ovarian mitochondrial dysfunctions. (a) Transcript levels of mitochondrial electron transport chain complex genes (CI, Ndufv1; CII, Sdhb; CIII, Uqcrc2; CV, Atp5a1) in the ovaries detected by real‐time RT‐PCR from 8‐month‐old WT, Ido1 −/− and Qprt −/− mice (n = 4 mice per group). (b) Western blot analysis of mitochondrial electron transport chain complex protein expression (CI, NDUFB8; CII, SDHB; CIII, UQCRC2; CIV, MTCO1; CV, ATP5A) in ovaries from 8‐month‐old WT, Ido1 −/− and Qprt −/− mice. (c) Relative expression of each protein was assessed as a ratio to GAPDH levels in each lane (n = 3 mice for each group). (d) Transcription analysis of essential dynamic genes for mitochondrial fusion (Mfn1, and Mfn2, Opa1) and fission (Drp1 and Fis1) (n = 4 mice for each group) in the ovaries from 8‐month‐old WT, Ido1 −/− and Qprt −/− mice. (e) Immunoblotting for mitochondrial fusion and fission proteins for each group. (f) Relative expression of each protein was assessed as a ratio to GAPDH levels in each lane (n = 3 mice for each group). (g) ATP content of granulosa cells from 8‐month‐old WT, Ido1 −/− and Qprt −/− mice. (h) Mitochondrial distribution was assessed by staining Mito‐tracker in the oocytes from 8‐month‐old WT, Ido1 −/− and Qprt −/− mice. Scale bars, 10 μm. (i) The abnormal mitochondrial distribution pattern percentages in oocytes from 8‐month‐old WT, Ido1 −/− and Qprt −/− mice (n = 42 oocytes for WT mice, n = 34 oocytes for Ido1 −/− mice, n = 37 oocytes for Qprt −/− mice). (j) Mitochondrial membrane potential was determined using JC‐1 staining in oocytes from 8‐month‐old WT, Ido1 −/− and Qprt −/− mice. Red fluorescence indicated high mitochondrial membrane potential, and green signals indicated low mitochondrial membrane potential. Scale bar, 50 μm. (k) Average ratios of red to green fluorescence in oocytes from each group (n = 7 oocytes for WT mice, n = 8 oocytes for Ido1 −/− mice, n = 10 oocytes for Qprt −/− mice). *p < 0.05, **p < 0.01, ***p < 0.001.