Accumulation of deletions in MtDNA during tissue aging: analysis by long PCR.
Multiple deletions of mtDNA have not only been implicated in aging, but also in a wide variety of pathological conditions. The enzyme system used in long-PCR makes it possible to synthesize the entire mitochondrial genome (16.5 kb), exposing the multiple deletions in mtDNAs implicated in and, at least partially, responsible for these pathologies. But it is not the number or type of anomalous mtDNA that is crucial, rather it is their frequency relative to the number of intact copies of the mitochondrial genome. Our work exposes the necessity of quantitating the number of normal mitochondrial DNAs. The accuracy of the technique and the small sample size required permit one to detect multiple deletions, located in a specific organ, and simultaneously measure the fraction of intact molecules. This fraction can then be correlated with mitochondrial dysfunction to serve both as an indicator of tissue aging and a monitor of an impending myopathy.
Unité 38 de l'INSERM, Laboratoire de Biochimie médicale, Faculté de Médecine, Marseille, France.
P
P
Reynier
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Malthiery
D000368:Aged; D000369:Aged, 80 and over; D000375:Aging / Q000235:genetics*; D001483:Base Sequence; D002311:Cardiomyopathy, Dilated / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004272:DNA, Mitochondrial / Q000235:genetics*; D004259:DNA-Directed DNA Polymerase; D006801:Humans; D008875:Middle Aged; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D011111:Polymyalgia Rheumatica / Q000235:genetics; D017384:Sequence Deletion; D019914:Taq Polymerase
D017931:DNA Primers; D004272:DNA, Mitochondrial; D019914:Taq Polymerase; D004259:DNA-Directed DNA Polymerase