Biogerontology

, Volume 6, Issue 1, pp 39–47 | Cite as

Testing the “garbage” accumulation theory of ageing: mitotic activity protects cells from death induced by inhibition of autophagy

  • Yuri Stroikin
  • Helge Dalen
  • Ulf T. Brunk
  • Alexei Terman
Research article

Abstract

Imperfect autophagic degradation of oxidatively damaged macromolecules and organelles (so-called biological “garbage”) is considered an important contributor to ageing and consequent death of postmitotic (non-dividing) cells, such as neurons and cardiac myocytes. In contrast, proliferating cells apparently escape senescence by a continuous dilution and repair of damaged structures during division. Postmitotic ageing can be mimicked and studied in cultures of potentially dividing cells if their mitotic activity is inhibited. To test the “garbage accumulation” theory of ageing, we compared survival of density-dependent growth-arrested (confluent) and proliferating human fibroblasts and astrocytes following inhibition of autophagic sequestration with 3-methyladenine (3MA). Exposure of confluent fibroblast cultures to 3MA for two weeks resulted in a significantly increased proportion of dying cells compared to both untreated confluent cultures and dividing cells with 3MA-inhibited autophagy. Similar results were obtained when autophagic degradation was suppressed by the protease inhibitor leupeptin. In 3MA- or leupeptin-exposed cultures, dying cells were overloaded with undegraded autofluorescent material. The results support a key role of biological lysosomal “garbage” accumulation in the triggering of ageing and death of postmitotic cells, as well as the anti-ageing role of cell division.

Keywords

ageing apoptosis astrocytes autophagocytosis fibroblasts 3-methyladenine leupeptin mitosis 

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References

  1. Ames, BN, Gold, LS 1990Too many rodent carcinogens: mitogenesis increases mutagenesisScience249970971Google Scholar
  2. Beckman, KB, Ames, BN 1998The free radical theory of aging maturesPhysiol Rev78547581Google Scholar
  3. Bergamini, E, Cavallini, G, Donati, A, Gori, Z 2003The anti-ageing effects of caloric restriction may involve stimulation of macroautophagy and lysosomal degradation, and can be intensified pharmacologicallyBiomed Pharmacother57203208Google Scholar
  4. Blommaart, EF, Krause, U, Schellens, JP, Vreeling-Sindelarova, H, Meijer, AJ 1997The phosphatidylinositol 3-kinase inhibitors wortmannin and LY294002 inhibit autophagy in isolated rat hepatocytesEur J Biochem243240246Google Scholar
  5. Brunk, UT, Terman, A 2002Lipofuscin: mechanisms of age-related accumulation and influence on cell functionsFree Radic Biol Med33611619Google Scholar
  6. Brunk, UT, Terman, A 2002The mitochondrial-lysosomal axis theory of aging: accumulation of damaged mitochondria as a result of imperfect autophagocytosisEur J Biochem26919962002Google Scholar
  7. Burnet, FM 1973A genetic interpretation of ageingLancet2480483Google Scholar
  8. Cadenas, E, Davies, KJ 2000Mitochondrial free radical generation, oxidative stress, and agingFree Radic Biol Med29222230Google Scholar
  9. Campisi, J 1996Replicative senescence: an old lives’ tale?Cell84497500Google Scholar
  10. Campisi, J 2000Cancer, aging and cellular senescenceIn Vivo14183188Google Scholar
  11. Grey, AD 2002Bioremediation meets biomedicine: therapeutic translation of microbial catabolism to the lysosomeTrends Biotechnol20452455Google Scholar
  12. Fonager, J, Beedholm, R, Clark, BF, Rattan, SI 2002Mild stress-induced stimulation of heat-shock protein synthesis and improved functional ability of human fibroblasts undergoing aging in vitroExp Gerontol3712231228Google Scholar
  13. Gerland, LM, Genestier, L, Peyrol, S, Michallet, MC, Hayette, S, Urbanowicz, I, Ffrench, P, Magaud, JP, Ffrench, M 2004Autolysosomes accumulate during in vitro CD8(+) T-lymphocyte aging and may participate in induced death sensitization of senescent cellsExp Gerontol39789800Google Scholar
  14. Goldberg, AL 2003Protein degradation and protection against misfolded or damaged proteinsNature426895899Google Scholar
  15. Grune, T, Merker, K, Sandig, G, Davies, KJ 2003Selective degradation of oxidatively modified protein substrates by the proteasomeBiochem Biophys Res Commun305709718Google Scholar
  16. Harman, D 1956Aging: a theory based on free radical and radiation chemistryJ Gerontol211298300Google Scholar
  17. Hirsch, HR 1978The waste-product theory of aging: waste dilution by cell divisionMech Ageing Dev85162Google Scholar
  18. Ivy, GO, Schottler, F, Wenzel, J, Baudry, M, Lynch, G 1984Inhibitors of lysosomal enzymes: accumulation of lipofuscin-like dense bodies in the brainScience226985987Google Scholar
  19. Kirkwood, TB 1989DNA, mutations and agingMutat Res21917Google Scholar
  20. Leist, M, Jaattela, M 2001Four deaths and a funeral: from caspases to alternative mechanismsNat Rev Mol Cell Biol2589598Google Scholar
  21. Lockshin, RA, Zakeri, Z 2004Caspase-independent cell death?Oncogene2327662773Google Scholar
  22. Martinez, DE 1998Mortality patterns suggest lack of senescence in hydraExp Gerontol33217225Google Scholar
  23. Melendez, A, Talloczy, Z, Seaman, M, Eskelinen, EL, Hall, DH, Levine, B 2003Autophagy genes are essential for dauer development and life-span extension in C. elegansScience30113871391Google Scholar
  24. Orgel, LE 1973Ageing of clones of mammalian cellsNature243441445Google Scholar
  25. Petiot, A, Ogier-Denis, E, Blommaart, EF, Meijer, AJ, Codogno, P 2000Distinct classes of phosphatidylinositol 3′-kinases are involved in signaling pathways that control macroautophagy in HT-29 cellsJ Biol Chem275992998Google Scholar
  26. Sheldrake, AR 1974The ageing, growth and death of cellsNature250381385Google Scholar
  27. Sohal, RS, Sohal, BH 1991Hydrogen peroxide release by mitochondria increases during agingMech Ageing Dev57187202Google Scholar
  28. Terman, A 2001Garbage catastrophe theory of aging: imperfect removal of oxidative damage?Redox Rep61526Google Scholar
  29. Terman, A, Abrahamsson, N, Brunk, UT 1999Ceroid/lipofuscin-loaded human fibroblasts show increased susceptibility to oxidative stressExp Gerontol34755770Google Scholar
  30. Terman, A, Brunk, UT 1998Ceroid/lipofuscin formation in cultured human fibroblasts: the role of oxidative stress and lysosomal proteolysisMech Ageing Dev104277291Google Scholar
  31. Terman A and Brunk UT (2004) Aging as a catabolic malfunction. Int J Biochem Cell Biol 36 (in press)Google Scholar
  32. Terman, A, Dalen, H, Brunk, UT 1999Ceroid/lipofuscin-loaded human fibroblasts show decreased survival time and diminished autophagocytosis during amino acid starvationExp Gerontol34943957Google Scholar
  33. Terman, A, Dalen, H, Eaton, JW, Neuzil, J, Brunk, UT 2003Mitochondrial recycling and aging of cardiac myocytes: the role of autophagocytosisExp Gerontol38863876Google Scholar
  34. Verbeke, P, Fonager, J, Clark, BF, Rattan, SI 2001Heat shock response and ageing: mechanisms and applicationsCell Biol Int25845857Google Scholar
  35. Weber, GF, Daley, J, Kraeft, SK, Chen, LB, Cantor, H 1997Measurement of apoptosis in heterogenous cell populationsCytometry27136144Google Scholar

Copyright information

© Springer 2005

Authors and Affiliations

  • Yuri Stroikin
    • 1
  • Helge Dalen
    • 1
  • Ulf T. Brunk
    • 2
  • Alexei Terman
    • 1
  1. 1.Division of Experimental Pathology II Faculty of Health SciencesLinköping UniversityLinköpingSweden
  2. 2.Department of Pharmacology, Faculty of Health SciencesLinköping UniversityLinköpingSweden

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