Time-course of mitochondrial gene expressions in mice brains: implications for mitochondrial dysfunction, oxidative damage, and cytochrome c in aging

J Neurochem. 2005 Feb;92(3):494-504. doi: 10.1111/j.1471-4159.2004.02884.x.

Abstract

The study of aging is critical for a better understanding of many age-related diseases. The free radical theory of aging, one of the prominent aging hypotheses, holds that during aging, increasing reactive oxygen species in mitochondria causes mutations in the mitochondrial DNA and damages mitochondrial components, resulting in senescence. Understanding a mitochondrial gene expression profile and its relationship to mitochondrial function becomes an important step in understanding aging. The objective of the present study was to determine mRNA expression of mitochondrial-encoded genes in brain slices from C57BL6 mice at four ages (2, 12, 18, and 24 months) and to determine how these altered mitochondrial genes influence age-related changes, including oxidative damage and cytochrome c in apoptosis. Using northern blot analysis, in situ hybridization, and immunofluorescence analyses, we analyzed changes in the expression of mitochondrial RNA encoding the mitochondrial genes, oxidative damage marker, 8-hydroxyguanosine (8-OHG), and cytochrome c in brain slices from the cortex of C57BL6 mice at each of the four ages. Our northern blot analysis revealed an increased expression of mitochondrial-encoded genes in complexes I, III, IV, and V of the respiratory chain in 12- and 18-month-old C57BL6 mice compared to 2-month-old mice, suggesting a compensatory mechanism that allows the production of proteins involved in the electron transport chain. In contrast to the up-regulation of mitochondrial genes in 12- and 18-month-old C57BL6 mice, mRNA expression in 24-month-old C57BL6 mice was decreased, suggesting that compensation maintained by the up-regulated genes cannot be sustained and that the down-regulation of expression results in the later stage of aging. Our in situ hybridization analyses of mitochondrial genes from the hippocampus and the cortex revealed that mitochondrial genes were over-expressed, suggesting that these brain areas are critical for mitochondrial functions. Our immunofluorescence analysis of 8-OHG and cytochrome c revealed increased 8-OHG and cytochrome c in 12-month-old C57BL6 mice, suggesting that age-related mitochondrial oxidative damage and apoptosis are associated with mitochondrial dysfunction. Our double-labeling analysis of in situ hybridization of ATPase 6 and our immunofluorescence analysis of 8-OHG suggest that specific neuronal populations undergo oxidative damage. Further, double-labeling analysis of in situ hybridization of ATPase 6 and immunofluorescence analysis of cytochrome c suggest cytochrome c release is related to mitochondrial dysfunction in the aging C57BL6 mouse brain. This study also suggests that these mitochondrial gene expression changes may relate to the role of mitochondrial dysfunction, oxidative damage, and cytochrome c in aging and in age-related diseases such as Alzheimer's disease and Parkinson's disease.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Aging / genetics*
  • Aging / metabolism
  • Animals
  • Blotting, Northern
  • Brain / metabolism*
  • Cytochromes c / genetics*
  • Cytochromes c / metabolism
  • Electron Transport Complex I / genetics
  • Electron Transport Complex III / genetics
  • Electron Transport Complex IV / genetics
  • Fluorescent Antibody Technique
  • Gene Expression Regulation, Developmental*
  • Guanosine / analogs & derivatives*
  • Guanosine / metabolism
  • In Situ Hybridization
  • Mice
  • Mice, Inbred C57BL
  • Mitochondria / genetics*
  • Mitochondria / metabolism
  • Mitochondrial Proton-Translocating ATPases / genetics
  • Oxidative Stress / genetics*
  • Protein Subunits / genetics
  • RNA / metabolism
  • RNA, Messenger / metabolism
  • RNA, Mitochondrial
  • Time

Substances

  • Protein Subunits
  • RNA, Messenger
  • RNA, Mitochondrial
  • Guanosine
  • 8-hydroxyguanosine
  • RNA
  • Cytochromes c
  • Electron Transport Complex IV
  • Mitochondrial Proton-Translocating ATPases
  • Electron Transport Complex I
  • Electron Transport Complex III
  • mt-Atp6 protein, mouse