Mitochondrial DNA rearrangements in aging human brain and in situ PCR of mtDNA.
Deletions of the mitochondrial DNA (mtDNA) have been shown to accumulate with age in a variety of species regardless of mean or maximal life span. This implies that such mutations are either a molecular biomarker of senescence or that they are more causally linked to senescence itself. One assay that can be used to detect these mtDNA mutations is the long-extension polymerase chain reaction assay. This assay amplifies approximately 16 kb of the mtDNA in mammalian mitochondria and preferentially amplifies mtDNAs that are either deleted or duplicated. We have applied this assay to the aging human brain and found a heterogeneous array of rearranged mtDNAs. In addition, we have developed in situ polymerase chain reaction to detect mtDNA within individual cells of both the mouse and the human brain as a first step in identifying and enumerating cells containing mutant mtDNAs in situ.
Melov
S
S
Center For Molecular Medicine, Emory University, Atlanta, GA 30322, USA. smelov@buckcenter.org
Schneider
J A
JA
Coskun
P E
PE
Bennett
D A
DA
Wallace
D C
DC
D000368:Aged; D000375:Aging / Q000235:genetics*; D000818:Animals; D002531:Cerebellum / Q000378:metabolism; D017931:DNA Primers; D004272:DNA, Mitochondrial / Q000235:genetics*; D004734:Energy Metabolism / Q000502:physiology; D005260:Female; D005625:Frontal Lobe / Q000378:metabolism; D017353:Gene Deletion; D015321:Gene Rearrangement; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008928:Mitochondria / Q000201:enzymology* / Q000235:genetics; D000226:Mitochondrial ADP, ATP Translocases / Q000235:genetics / Q000378:metabolism; D018384:Oxidative Stress / Q000502:physiology; D016133:Polymerase Chain Reaction / Q000379:methods; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism
D017931:DNA Primers; D004272:DNA, Mitochondrial; D013481:Superoxides; D000226:Mitochondrial ADP, ATP Translocases; D013482:Superoxide Dismutase