Journal
REPRODUCTIVE BIOMEDICINE ONLINE
Volume 17, Issue 6, Pages 806-813Publisher
ELSEVIER SCI LTD
DOI: 10.1016/S1472-6483(10)60409-9
Keywords
embryo; fetus; mitochondria; mtDNA; reactive oxygen species
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Funding
- Elmore Fund
- Trinity College, Cambridge
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Mitochondria are inherited maternally via the oocyte, which in the mouse contains 150-250 x 10(3) 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.
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