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Measurement of Caspase Activity: From Cell Populations to Individual Cells

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Mammalian Cell Viability

Part of the book series: Methods in Molecular Biology ((MIMB,volume 740))

Abstract

Caspases are critical regulators of the apoptotic program, responsible for the harmonic dismantling of the cell. Cell death can occur by way of different options (necroptosis, necrosis, extreme autophagy) but once caspases are fully engaged it will take the apoptotic route. Hence, in general, caspase activity is inversely proportional to cell viability. Caspase activation can be measured by means of different techniques. Here, we describe three different methodologies to measure/observe caspase activation. Two approaches are recommended for studies on the whole cell population, whereas the third one is designed for visualizing caspase activation in single live cells.

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References

  1. Caroline, H. Y., and Yuan, J. (2009) The Jekyll and Hyde Functions of Caspases Dev. Cell 16, 21–34

    Google Scholar 

  2. Zong W.X., and Thompson, C.B. (2006) Necrotic death as a cell fate. Genes Dev. 20, 1–15

    Article  PubMed  CAS  Google Scholar 

  3. Pop, C., and Salvesen, G.S. (2009) Human caspases: activation, specificity, and regulation. J Biol Chem. 284, 21777–81

    Article  PubMed  CAS  Google Scholar 

  4. Paroni, G., Henderson, C., Schneider, C., and Brancolini, C. (2001) Caspase-2-induced apoptosis is dependent on caspase-9, but its processing during UV- or tumor necrosis factor-dependent cell death requires caspase-3. J Biol Chem. 276, 21907–1

    Article  PubMed  CAS  Google Scholar 

  5. Rodriguez J, Lazebnik Y. (1999) Caspase-9 and APAF-1 form an active holoenzyme. Genes Dev. 13, 3179–84

    Article  PubMed  CAS  Google Scholar 

  6. Henderson, C.J., Aleo, E., Fontanini, A., Maestro, R., Paroni, G., and Brancolini C. (2005) Caspase activation and apoptosis in response to proteasome inhibitors. Cell Death Differ. 12, 1240–54

    Article  PubMed  CAS  Google Scholar 

  7. Lüthi, A.U., and Martin, S.J. (2007) The CASBAH: a searchable database of caspase substrates. Cell Death Differ. 14, 641–50

    Article  PubMed  Google Scholar 

  8. Garcia-Calvo, M., Peterson, E.P., Leiting, B., Ruel, R., Nicholson, D.W., and Thornberry, N.A. (1998) Inhibition of human caspases by peptide-based and macromolecular inhibitors. J Biol Chem. 273, 32608–13

    Article  PubMed  CAS  Google Scholar 

  9. Rehm, M., Dussmann, H., Janicke, R.U., Tavare, J.M., Kogel, D., Prehn, J.H. (2002). Single-cell fluorescence resonance energy transfer analysis demonstrates that caspase activation during apoptosis is a rapid process. Role of caspase-3. J Biol Chem. 277, 24506–14

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This work is supported by AIRC (Associazione Italiana Ricerca sul Cancro), MIUR (Ministero dell’Istruzione, Università e Ricerca), and Regione Friuli-Venezia Giulia (AITT-LR25/07).

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Correspondence to Claudio Brancolini .

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Paroni, G., Brancolini, C. (2011). Measurement of Caspase Activity: From Cell Populations to Individual Cells. In: Stoddart, M. (eds) Mammalian Cell Viability. Methods in Molecular Biology, vol 740. Humana Press. https://doi.org/10.1007/978-1-61779-108-6_9

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  • DOI: https://doi.org/10.1007/978-1-61779-108-6_9

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-107-9

  • Online ISBN: 978-1-61779-108-6

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