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Causes and Consequences of Damage to Mitochondria

Study of Functional Aspects by Flow Cytometry

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Book cover Aging Methods and Protocols

Part of the book series: Methods in Molecular Medicine ((MIMM,volume 38))

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Abstract

A rapidly increasing amount of data supports the view that progressive bioenergetic loss caused by injury of the main energy-producing subcellular organelles, that is, the mitochondria, plays a key role in aging. A link between senescence and energy loss is already implied in Harman’s (1) free radical theory of aging, according to which oxygen-derived free radicals injure the cells, with concomitant impairment of performance at the cellular and physiological levels. Further, Miquel and co-workers (2, 3) have proposed a mitochondrial theory of aging, according to which aging results from oxygen stress damage to the mitochondrial genome, with concomitant bioenergetic decline. More recently, a number of laboratories, including our own (46), have provided biochemical data in agreement with the above views. Thus, we have shown that, as the result of age-related oxygen stress, mitochondrial glutathione is oxidized in direct relation to injury of the mitochondrial DNA (5). Further, our studies suggest that an antioxidant product extracted from Ginkgo biloba may counteract in part the damaging effects of free radicals on mitochondrial and cellular aging (7).

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References

  1. Harman, D. (1956) Aging: a theory based on free radical and radiation chemistry. J. Gerontol. 11, 298–300.

    PubMed  CAS  Google Scholar 

  2. Miquel, J. and Fleming, J. (1986) Theoretical and experimental support for an “oxygen radical-mitochondrial injury hypothesis of cell aging, in Free Radicals, Aging and Degenerative Diseases (Johnson, J. E., Jr., Walford, R., Harman, D., and Miquel, J., eds.) Alan R. Liss, New York, pp. 51–74.

    Google Scholar 

  3. Miquel, J. (l998) An update on the oxygen stress-mitochondrial mutation theory of aging: genetic and evolutionary implications. Exp. Gerontol. 33, 113–126.

    Article  PubMed  CAS  Google Scholar 

  4. Yen, T. C., Chen, Y. S., King, K. L., Yeh, S. H., and Wei, Y. H. (1989) Liver mitochondrial respiratory functions decline with age. Biochem. Biophys. Res. Commun. 165, 994–1003.

    Article  Google Scholar 

  5. García de la Asunción, J., Millán, A., Pla, R., Bruseghini, L., Esteras, A., Pallardo, F. V., Sastre, J., and Viña, J. (1996) Aging of the liver: age associated oxidative damage to mitochondrial DNA. FASEB J. 10, 333–338.

    Google Scholar 

  6. Sastre, J., Pallardo, F. V., Pla, R., Pellin, A., Juan, G., O’Connor, E., Estrela, J. M., Miquel, J., and Viña, J. (1996) Aging of the liver: age associated mitochondrial damage in intact hepatocytes. Hepatology 24, 1199–1205.

    Article  PubMed  CAS  Google Scholar 

  7. Sastre, J., Millan, A., García de la Asunción, J., Pla, R., Juan, G., Pallardó, F. V., O’Connor, E., Martin, J. A., Droix-Lefaix, M. T., and Viña, J. (1998) A Ginkgo biloba extract (Egb 761) prevents mitochondrial aging by protecting against oxidative stress. Free Radic. Biol. Med. 24, 298–304.

    Article  PubMed  CAS  Google Scholar 

  8. Hagen, T. M., Yowe, D. L., Bartholomew, J. C., Wehr, C. M., Do, K. L., Park, J.-Y., and Ames, B. N. (1997) Mitochondrial decay in hepatocytes from old rats: membrane potential declines, heterogeneity and oxidants increase. Proc. Natl. Acad. Sci. USA 94, 3064–3069.

    Article  PubMed  CAS  Google Scholar 

  9. Chen, L. B. (1989) Mitochondrial membrane potential in living cells. Ann. Rev. Cell Biol. 4, 155–181.

    Article  Google Scholar 

  10. Darley-Usmar, V. M., Rickwood, D., and Wilson, M. T., eds. (1987) Mitochondria: a Practical Approach, IRL, Oxford, pp. 1–16.

    Google Scholar 

  11. Ferrandiz, M. L., Martínez, M., De Juan, E., Díez, A., Bustos, G. and Miquel, J. (1994) Impairment of mitochondrial oxidative phosphorylation in the brain of aged mice. Brain Res. 644, 335–338.

    Article  PubMed  CAS  Google Scholar 

  12. Martínez, M., Ferrándiz, M. L., Díez, A., and Miquel, J. (1995) Depletion of cytosolic GSH decreases the ATP levels and viability of synaptosomes from aged mice but not from young mice. Mech. Ageing Dev. 84, 77–81.

    Article  PubMed  Google Scholar 

  13. Maftah, A., Petit, J. M., Ratinaud, M. H., and Julien, R. (1989) 10-N nonyl-acridine orange: a fluorescent probe which stains mitochondria independently of their energetic state. Biochem. Biophys. Res. 164, 185–190.

    Article  CAS  Google Scholar 

  14. Petit, P. X., O’Connor, J. E., Grunawald, D., and Brown, S. C. (1990) Analysis of the membrane potential of rat-and mouse-liver mitochondria by flow cytometry and possible implications. Eur. J. Biochem. 194, 389–397.

    Article  PubMed  CAS  Google Scholar 

  15. Greenspan, P., Mayer, E. P., and Fowler, S. D. (1985) Nile red: a selective fluorescent stain for intracellular lipid droplets. J. Cell Biol. 100, 965–973.

    Article  PubMed  CAS  Google Scholar 

  16. Royal, J. A. and Ischiripoulos, H. (1993) Evaluation of 2′–7′-dichloro intracellular H2O2 in cultured endothelial cells. Arch. Biochem. Biophys. 302, 348–355, 1993.

    Article  Google Scholar 

  17. Miquel, J., Economos, A. C., and Bensch, K. G. (1981) Insect vs. mammalian aging, in Aging and Cell Structure (Johnson, J. E. Jr., ed.), Plenum Press, New York, pp.347–379.

    Google Scholar 

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© 2000 Humana Press Inc., Totowa, NJ

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Pallardo, F.V., Sastre, J., Miquel, J., Viña, J. (2000). Causes and Consequences of Damage to Mitochondria. In: Barnett, Y.A., Barnett, C.R. (eds) Aging Methods and Protocols. Methods in Molecular Medicine, vol 38. Humana Press. https://doi.org/10.1385/1-59259-070-5:237

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  • DOI: https://doi.org/10.1385/1-59259-070-5:237

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-582-9

  • Online ISBN: 978-1-59259-070-4

  • eBook Packages: Springer Protocols

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