Abstract
Countless studies have implicated reactive oxygen species (ROS) and mitochondrial dysfunction in the ageing process. During cellular senescence, the ultimate and irreversible loss of replicative capacity of somatic cells grown in culture, several studies have reported increased levels of ROS associated with mitochondrial dysfunction and metabolic inefficiency. Moreover, studies have revealed that interventions modulating intracellular ROS can impact on the replicative lifespan of cultured cells, suggesting that ROS play a central role in the process. In this chapter, we present several protocols used for detection of (intra- and extracellular) ROS in live cells.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Balaban RS, Nemoto S, Finkel T (2005) Mitochondria, oxidants, and aging. Cell 120:483–495
Passos JF, von Zglinicki T (2005) Mitochondria, telomeres and cell senescence. Exp Gerontol 40:466–472
von Zglinicki T (2002) Oxidative stress shortens telomeres. Trends Biochem Sci 27:339–344
Ahmed EK, Rogowska-Wrzesinska A, Roepstorff P, Bulteau A-L, Friguet B (2010) Protein modification and replicative senescence of WI-38 human embryonic fibroblasts. Aging Cell 9:252–272
Sitte N, Merker K, von Zglinicki T, Grune T (2000) Protein oxidation and degradation during proliferative senescence of human MRC-5 fibroblasts. Free Radic Biol Med 28:701–708
Sitte N, Merker K, Grune T, von Zglinicki T (2001) Lipofuscin accumulation in proliferating fibroblasts in vitro: an indicator of oxidative stress. Exp Gerontol 36:475–486
Passos JF, Saretzki G, Ahmed S, Nelson G, Richter T, Peters H, Wappler I, Birkett M, Harold G, Schaeuble K, Birch-Machin M, Kirkwood T, von Zglinicki T (2007) Mitochondrial dysfunction accounts for the stochastic heterogeneity in telomere-dependent senescence. PLoS Biol 5:e110
Allen RG, Tresini M, Keogh BP, Doggett DL, Cristofalo VJ (1999) Differences in electron transport potential, antioxidant defenses, and oxidant generation in young and senescent fetal lung fibroblasts (WI-38). J Cell Physiol 180:114–122
Hutter E, Renner K, Pfister G, Stockl P, Jansen-Durr P, Gnaiger E (2004) Senescence-associated changes in respiration and oxidative phosphorylation in primary human fibroblasts. Biochem J 380:919–928
Passos JF, Nelson G, Wang C, Richter T, Simillion C, Proctor CJ, Miwa S, Olijslagers S, Hallinan J, Wipat A, Saretzki G, Rudolph KL, Kirkwood TBL, von Zglinicki T (2010) Feedback between p21 and reactive oxygen production is necessary for cell senescence. Mol Syst Biol 6:347
Serra V, von Zglinicki T, Lorenz M, Saretzki G (2003) Extracellular superoxide dismutase is a major antioxidant in human fibroblasts and slows telomere shortening. J Biol Chem 278:6824–6830
Saretzki G, Murphy MP, von Zglinicki T (2003) MitoQ counteracts telomere shortening and elongates lifespan of fibroblasts under mild oxidative stress. Aging Cell 2:141–143
Petersen S, Saretzki G, von Zglinicki T (1998) Preferential accumulation of single-stranded regions in telomeres of human fibroblasts. Exp Cell Res 239:152–160
Lee AC, Fenster BE, Ito H, Takeda K, Bae NS, Hirai T, Yu Z-X, Ferrans VJ, Howard BH, Finkel T (1999) Ras proteins induce senescence by altering the intracellular levels of reactive oxygen species. J Biol Chem 274:7936–7940
Macip S, Igarashi M, Berggren P, Yu J, Lee SW, Aaronson SA (2003) Influence of induced reactive oxygen species in p53-mediated cell fate decisions. Mol Cell Biol 23:8576–8585
Macip S, Igarashi M, Fang L, Chen A, Pan Z, Lee S, Aaronson S (2002) Inhibition of p21-mediated ROS accumulation can rescue p21-induced senescence. EMBO J 21:2180–2188
Takahashi A, Ohtani N, Yamakoshi K, Iida S, Tahara H, Nakayama K, Nakayama KI, Ide T, Saya H, Hara E (2006) Mitogenic signalling and the p16(INK4a)-Rb pathway cooperate to enforce irreversible cellular senescence. Nat Cell Biol 8:1291–1297
Colavitti R, Finkel T (2005) Reactive oxygen species as mediators of cellular senescence. IUBMB Life 57:277–281
de Magalhaes JP, Church GM (2006) Cells discover fire: employing reactive oxygen species in development and consequences for aging. Exp Gerontol 41:1–10
Kalyanaraman B, Darley-Usmar V, Davies KJA, Dennery PA, Forman HJ, Grisham MB, Mann GE, Moore K, Roberts Ii LJ, Ischiropoulos H (2011) Measuring reactive oxygen and nitrogen species with fluorescent probes: challenges and limitations. Free Radic Biol Med 52:1–6
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Busincess Media, LLC
About this protocol
Cite this protocol
Passos, J.F., Miwa, S., von Zglinicki, T. (2013). Measuring Reactive Oxygen Species in Senescent Cells. In: Galluzzi, L., Vitale, I., Kepp, O., Kroemer, G. (eds) Cell Senescence. Methods in Molecular Biology, vol 965. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-239-1_17
Download citation
DOI: https://doi.org/10.1007/978-1-62703-239-1_17
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-238-4
Online ISBN: 978-1-62703-239-1
eBook Packages: Springer Protocols