Mitochondrial DNA mutation analysis in human skin fibroblasts from fetal, young, and old donors

Glenn S. Gerhard, Floyd A. Benko, R. G. Allen, Maria Tresini, Anne Kalbach, Vincent J. Cristofalo, Christopher Gocke

Research output: Contribution to journalArticle

Abstract

Multibase deletions in mitochondrial DNA (mtDNA) have been shown to accumulate with age in several tissues, including skin, whereas point mutations have only recently been demonstrated to increase during aging, with several specific mutations occurring at high levels (up to 50%) in skin fibroblasts obtained from old donors [Science 286(1999)774]. We have conducted a survey for a specific deletion and for point mutations in several regions of mtDNA from cultured skin fibroblasts derived from eight fetal (12-20 weeks gestational age), ten young (17-33 years of age) and 11 old (78-92 years of age) human donors. Using PCR analysis, detectable levels of the 4977 basepair (bp) 'common deletion' were present in all three age groups, with the highest deletion levels of up to 0.3% of total mtDNA found in several cell lines from old donors, although other old donor cell lines had much lower levels. Single strand conformation polymorphism (SSCP) analysis for point mutations in the non-coding D-loop region and two regions of the cytochrome oxidase 2 gene failed to reveal the presence of any single base mutations. We infer that age-related high level mutational damage in mtDNA from human skin fibroblasts may manifest both sequence and inter-individual specificity.

Original languageEnglish (US)
Pages (from-to)155-166
Number of pages12
JournalMechanisms of Ageing and Development
Volume123
Issue number2-3
DOIs
StatePublished - 2002
Externally publishedYes

Fingerprint

Fibroblasts
Mitochondrial DNA
Skin
Point Mutation
Tissue Donors
Mutation
Cells
Cell Line
Sequence Deletion
Electron Transport Complex IV
Polymorphism
Gestational Age
Conformations
Age Groups
Genes
Aging of materials
Tissue
Polymerase Chain Reaction

Keywords

  • Aging
  • Mitochondrial DNA
  • Skin fibroblast

ASJC Scopus subject areas

  • Aging
  • Biochemistry
  • Developmental Biology
  • Developmental Neuroscience

Cite this

Mitochondrial DNA mutation analysis in human skin fibroblasts from fetal, young, and old donors. / Gerhard, Glenn S.; Benko, Floyd A.; Allen, R. G.; Tresini, Maria; Kalbach, Anne; Cristofalo, Vincent J.; Gocke, Christopher.

In: Mechanisms of Ageing and Development, Vol. 123, No. 2-3, 2002, p. 155-166.

Research output: Contribution to journalArticle

Gerhard, Glenn S. ; Benko, Floyd A. ; Allen, R. G. ; Tresini, Maria ; Kalbach, Anne ; Cristofalo, Vincent J. ; Gocke, Christopher. / Mitochondrial DNA mutation analysis in human skin fibroblasts from fetal, young, and old donors. In: Mechanisms of Ageing and Development. 2002 ; Vol. 123, No. 2-3. pp. 155-166.
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