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Assessing Mitochondrial Selective Autophagy in the Nematode Caenorhabditis elegans

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Mitochondria

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1567))

Abstract

Eukaryotic cells heavily depend on ATP generated by oxidative phosphorylation (OXPHOS) within mitochondria. Besides being the main suppliers of cell’s energy, mitochondria also provide an additional compartment for a wide range of cellular processes and metabolic pathways. Mitochondria constantly undergo fusion/fission events and form a mitochondrial network, which is a highly dynamic, tubular structure allowing for rapid and continuous exchange of genetic material, as well as, targeting dysfunctional mitochondria for degradation through mitochondrial selective autophagy (mitophagy). Mitophagy mediates the elimination of damaged and/or superfluous organelles, maintaining mitochondrial and cellular homeostasis. In this chapter, we present two versatile, noninvasive methods, developed for monitoring in vivo mitophagy in C. elegans. These procedures enable the assessment of mitophagy in several cell types during development or under stress conditions. Investigating the role of mitophagy at the organismal level is essential for the development of therapeutic interventions against age-related diseases.

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Abbreviations

CCCP:

Carbonyl cyanide m-chlorophenylhydrazone

DMSO:

Dimethyl sulfoxide

DsRed:

Red fluorescent protein

FUdR:

Fluorodeoxyuridine

GFP:

Green fluorescent protein

MAP1LC3/LC3:

Microtubule-associated protein 1 light chain 3

mtGFP:

Mitochondria-targeted green fluorescent protein

mtRosella:

Mitochondria-targeted Rosella

NGM:

Nematode growth medium

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Acknowledgments

We thank A. Pasparaki for expert technical support. We thank R. Devenish for providing the pAS1NB-CS-Rosella plasmid. Some nematode strains used in this work were provided by the Caenorhabditis Genetics Center, which is funded by the National Center for Research Resources (NCRR) of the National Institutes of Health (NIH). We thank A. Fire for plasmid vectors. This work was funded by grants from the European Research Council (ERC) and the European Commission 7th Framework Programme.

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Authors and Affiliations

  1. Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology Hellas, N. Plastira 100, Vassilika Vouton, 1385, Heraklion, 70013, Crete, Greece

    Konstantinos Palikaras & Nektarios Tavernarakis

  2. Faculty of Medicine, Department of Basic Sciences, University of Crete, Heraklion, 71003, Crete, Greece

    Nektarios Tavernarakis

Authors
  1. Konstantinos Palikaras
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  2. Nektarios Tavernarakis
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Corresponding author

Correspondence to Nektarios Tavernarakis .

Editor information

Editors and Affiliations

  1. Biomedical Center-Physiological Chemistry, LMU Munich, Martinsried, Germany

    Dejana Mokranjac

  2. Gene Center, LMU Munich and Institute for Diabetes and Obesity, Helmholtz Zentrum München, Munich, Germany

    Fabiana Perocchi

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Palikaras, K., Tavernarakis, N. (2017). Assessing Mitochondrial Selective Autophagy in the Nematode Caenorhabditis elegans . In: Mokranjac, D., Perocchi, F. (eds) Mitochondria. Methods in Molecular Biology, vol 1567. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6824-4_21

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  • DOI: https://doi.org/10.1007/978-1-4939-6824-4_21

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6822-0

  • Online ISBN: 978-1-4939-6824-4

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