Variations in mouse mitochondrial DNA copy number from fertilization to birth are associated with oxidative stress

Mitochondria are inherited maternally via the oocyte, which in the mouse contains 150–250 × 103 copies of mitochondrial DNA (mtDNA). The number of mtDNA copies/embryo is thought to be stable during cleavage, being progressively diluted/cell with each round of cell division, until replication begins at an undefined time post-implantation. Post-natally, tissues differ in copy number of mtDNA/cell, but when and how these differences arise is unclear. A ratiometric quantitative real-time polymerase chain reaction assay of the levels of a single mitochondrial gene against a single copy nuclear gene was used to estimate the average copy value of mtDNA/per cell from zygote to birth. A novel Bayesian statistical model was used to identify day 5.15–6.15 as the time at which replication recommences, consistent with the viability patterns of embryos carrying mitochondrial mutations. Mitochondrial DNA copy number/cell in a range of post-day 9.5 fetal and placental tissues showed tissue-specific temporal expression patterns. Western blotting was used to quantify post-day 9.5 tissue markers for oxidative stress and manganese superoxide dismutase, and revealed correlations with the changes in mtDNA copy number. These findings have potential implications for fetal programming, in-vitro embryo culture, and the mechanism underlying the mitochondrial bottleneck.