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Identification and Characterization of Programmed Cell Death Markers in Bacterial Models

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Necrosis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1004))

Abstract

In eukaryotic organisms facing terminal stress, activation of genetically encoded cell death pathways underlies fundamental changes in core cellular processes and functional modification of critical biomolecules. These physiological alterations manifest themselves as phenotypic hallmarks during programmed cell death, and are markers of the particular mode of death initiated. A growing volume of work has illustrated that prokaryotes too are capable of exhibiting hallmarks of programmed cell death, albeit without the multiple, tight regulatory layers which control these events in higher order organisms.

This chapter describes how methods and materials which have been used to assay for hallmarks of programmed cell death in eukaryotic models are transferrable to prokaryotic models. In particular, we describe the applicability of these methods to the study of post-antibiotic effects on bacteria, notably the biochemical changes induced by the interaction of drug molecules and targets, including oxidative stress, that accompany and ensure cell death. Specifically we discuss techniques for detecting DNA fragmentation, chromosomal condensation, phosphatidylserine exposure, membrane depolarization, and caspase substrate peptide binding, thereby providing a launchpoint for the study of the evolution of these physiological events in bacteria.

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Acknowledgments

We thank James Collins for his mentorship and support of this work through his Howard Hughes Medical Institute Investigator and NIH Director’s Pioneer Award Program awards. We also thank Jarred Callura for his assistance with UV irradiation treatment design and Ahmad Khalil for his assistance with fluorescent microscopy, as well as with the design of Qdot-based Annexin V experiments.

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Authors and Affiliations

  1. Department of Biomedical Engineering and Center for BioDynamics, Howard Hughes Medical Institute, Boston University, Boston, MA, USA

    Daniel J. Dwyer & Jonathan A. Winkler

Authors
  1. Daniel J. Dwyer
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  2. Jonathan A. Winkler
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Editor information

Editors and Affiliations

  1. Department of Biology, Boston University, Boston, MA, USA

    Kimberly McCall

  2. Brookline, MA, USA

    Charles Klein

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Dwyer, D.J., Winkler, J.A. (2013). Identification and Characterization of Programmed Cell Death Markers in Bacterial Models. In: McCall, K., Klein, C. (eds) Necrosis. Methods in Molecular Biology, vol 1004. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-383-1_11

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  • DOI: https://doi.org/10.1007/978-1-62703-383-1_11

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-382-4

  • Online ISBN: 978-1-62703-383-1

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