How is mtDNA damage measured?
The quantitative polymerase chain reaction (QPCR) assay allows measurement of DNA damage in the mitochondrial and nuclear genomes without isolation of mitochondria. It also permits measurement of relative mitochondrial genome copy number.
How do you calculate mitochondrial DNA?
To determine the mitochondrial DNA content, relative to nuclear DNA use the following equations:
- ΔCT = (nucDNA CT – mtDNA CT)
- Relative mitochondrial DNA content = 2 × 2. ΔCT
What is mtDNA depletion?
Abstract. Mitochondrial DNA (mtDNA) depletion syndromes (MDS) are a genetically and clinically heterogeneous group of autosomal recessive disorders that are characterized by a severe reduction in mtDNA content leading to impaired energy production in affected tissues and organs.
What is the mutation rate for mitochondrial DNA?
The most recent estimations of the human germline mtDNA mutation rate are 1.30 × 10–8 21 or 1.89 × 10–8 22 mutations per site per year (assuming a generation time of 25 years). Consequently, we are using here an average rate of mutation success of 1.947 × 10–4 per genome per year.
What is the link between mitochondria and DNA damage?
Moreover, mitochondria contain inadequate DNA repair pathways, and, diminished DNA repair capacity may be one of the factors responsible for high mutation frequency of the mtDNA. mtDNA damage might cause impaired mitochondrial function, and, unrepaired mtDNA damage has been frequently linked with several diseases.
How many copies of mitochondrial DNA are in mitochondria?
Each mitochondrion can contain 2–10 copies of mtDNA and up to 1000 mitochondria are present per cell3.
Why mtDNA mutation rate is high?
In most metazoans, mtDNA shows an elevated mutation rate compared with nuclear DNA, likely due to less efficient DNA repair, a more mutagenic local environment (putatively caused by oxidative radicals), and an increased number of replications per cell division (Birky 2001; reviewed in Lynch 2007).
Why is mitochondrial DNA prone to damage?
Because mtDNA is deficient in histones, and therefore lacks the complexity associated with nuclear DNA, mtDNA may be more prone to attack by oxidants.