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- Title
Digital PCR Quantitation of Muscle Mitochondrial DNA: Age, Fiber Type, and Mutation-Induced Changes.
- Authors
Herbst, Allen; Widjaja, Kevin; Nguy, Beatrice; Lushaj, Entela B.; Moore, Timothy M.; Hevener, Andrea L.; McKenzie, Debbie; Aiken, Judd M.; Wanagat, Jonathan
- Abstract
Definitive quantitation of mitochondrial DNA (mtDNA) and mtDNA deletion mutation abundances would help clarify the role of mtDNA instability in aging. To more accurately quantify mtDNA, we applied the emerging technique of digital polymerase chain reaction to individual muscle fibers and muscle homogenates from aged rodents. Individual fiber mtDNA content correlated with fiber type and decreased with age. We adapted a digital polymerase chain reaction deletion assay that was accurate in mixing experiments to a mutation frequency of 0.03% and quantitated an age-induced increase in deletion frequency from rat muscle homogenates. Importantly, the deletion frequency measured in muscle homogenates strongly correlated with electron transport chain-deficient fiber abundance determined by histochemical analyses. These data clarify the temporal accumulation of mtDNA deletions that lead to electron chain-deficient fibers, a process culminating in muscle fiber loss.
- Subjects
MITOCHONDRIAL DNA; MUSCLES; POLYMERASE chain reaction; DELETION mutation; GENETICS of aging; DNA metabolism; AGE distribution; ANIMALS; DNA; MICE; MITOCHONDRIA; GENETIC mutation; RATS; SKELETAL muscle
- Publication
Journals of Gerontology Series A: Biological Sciences & Medical Sciences, 2017, Vol 72, Issue 10, p1327
- ISSN
1079-5006
- Publication type
journal article
- DOI
10.1093/gerona/glx058