Elimination of paternal mitochondria in mouse embryos occurs through autophagic degradation dependent on PARKIN and MUL1
- California Institute of Technology, United States
Figures
Figure 1 with 1 supplement
Paternal mitochondria are degraded by 84 hr after fertilization.
(A) Fluorescence of mito-Dendra2 in a live sperm cell isolated from the cauda epididymis of a PhAM mouse. (B–F) Mito-Dendra2 in a 12 hr (B), 36 hr (C), 60 hr (D), 72 hr (E), and 84 hr embryo (F). In …
Figure 1—figure supplement 1
Persistence of maternal versus paternal mitochondria after fertilization.
(A) Quantification of Figure 1H. Embryos were collected from crosses of homozygous mito-Dendra2 females with unlabeled males, or homozygous mito-Dendra2 males with unlabeled females. The total …
Figure 2
Induction of mitophagy by OXPHOS-inducing medium.
Mitophagy was examined in cells stably expressing Cox8-EGFP-mCherry. Wild-type (A) or Atg3 knockout mouse embryonic fibroblasts (MEFs) (B) were grown in Glucose (Glu) or Acetoacetate (Ac) containing …
Figure 3 with 1 supplement
Mitophagy under OXPHOS-inducing conditions requires FIS1, TBC1D15, and p62.
(A) Mitophagy in cells with mutations in mitochondrial dynamics genes. MEFs of the indicated genotype were cultured in glucose or acetoacetate medium, and mitophagy was quantified using the …
Figure 3—figure supplement 1
p62 knockout cells have defective OXPHOS-induced mitophagy.
(A) Quantification of red-only puncta in wild-type or p62 knockout cells grown in medium containing glucose (Glu) or acetoacetate (Ac). Error bars indicate SD, three biological replicates, p=0.0163 …
Figure 4 with 1 supplement
MUL1 and PARKIN have redundant functions in OXPHOS-induced mitophagy.
(A) Quantification of red-only puncta in cells grown in acetoacetate-containing medium. Presence (+) or absence (-) of Pink1, Parkin, or Mul1 is indicated. Error bars indicate SD of three biological …
Figure 4—figure supplement 1
Defective mitophagy in Parkin/Mul1-deficient cells.
(A) Requirement for Parkin/Mul1 in mitophagy. Quantification of red-only puncta in cells grown in medium containing acetoacetate. Wild-type (+) or Parkin knockout (-) cells were transduced with one …
Figure 5 with 1 supplement
Clearance of paternal mitochondria in preimplantation embryos requires mitophagy genes.
(A) Impaired elimination of paternal mitochondria upon inhibition of mitophagy genes. Embryos were injected with lentivirus expressing shRNA against the indicated genes. The mitochondrial Dendra2 …
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Figure 5—source data 1
Source data for Figure 5B and D.
Excel file containing source data for the plots in Figure 5B and D.
- https://doi.org/10.7554/eLife.17896.011
Figure 5—figure supplement 1
Inhibition of OXPHOS-induced mitophagy by dominant negative FIS1.
(A) Quantification of red-only puncta in wildtype (WT) cells or cells transduced with MYC-FIS1-DN retrovirus. Cells were grown in medium containing glucose (Glu) or acetoacetate (Ac). Error bars …
Figure 6 with 1 supplement
Loss of membrane potential in paternal mitochondria after fertilization.
(A) Mitochondrial membrane potential in live sperm cell. Spermatozoa were isolated from the cauda epididymis of a PhAM mouse, stained with 20 nM TMRE, washed, and imaged by fluorescent microscopy. …
Figure 6—figure supplement 1
Fusion activity of maternal mitochondria versus paternal mitochondria in the early embryo.
(A) Monitoring mitochondrial fusion in embryos. Embryos were collected from crosses of homozygous mito-Dendra females with unlabeled males (top panel), or homozygous mito-Dendra male with unlabeled …
