Journal
EXPERIMENTAL GERONTOLOGY
Volume 37, Issue 12, Pages 1389-1400Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/S0531-5565(02)00121-3
Keywords
single-cell PCR; mitochondrial DNA; astrocytes; neurons; 4977 bp deletion; aging
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Mitochondria are considered a key element in the process of organismic aging, because of their fundamental role in cellular energy generation. In the course of oxidative phosphorylation, harmful free radicals are continuously produced damaging the mitochondrial (mt) genome. One of the consequences is the occurrence of large-scale deletions in mtDNA molecules. The 4977 bp common deletion accumulates exponentially with age, in a mosaic pattern, especially in postmitotic tissues. In order to investigate whether certain cell characteristics underlie this pattern of distribution, and to look for possible age-related changes, two cell types in the caudate nucleus of the human brain from five young and five senescent subjects were analysed by single-cell PCR. MAP2-positive neurons and GFAP-positivc astrocytes were isolated by micromanipulation. For each of the 10 cases, at least 30 cells of each type were collected and subjected to PCR individually. Screening for the presence of the common deletion yielded no significant differences in relative distribution, neither between astrocytes and neurons, nor between healthy young and old humans. Our results imply that the age-dependent increase of the common deletion cannot come about by an increase of independent deletion events in a greater proportion of cells, and that mitotic rate is not a major cellular risk factor for deletion accumulation in the caudate nucleus. (C) 2002 Elsevier Science Inc. All rights reserved.
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