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Method Article
Nektarios Tavernarakis
Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology - Hellas, Department of Basic Sciences, Faculty of Medicine, University of Crete, Heraklion 71110, Crete, Greece; Tavernarakis lab
Corresponding Author
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Mitochondrial dysfunction is a shared hallmark of many human pathologies and ageing. Mitochondrial selective autophagy mediates the removal of damaged or superfluous mitochondria preserving mitochondrial and cellular homeostasis. Deregulation of mitophagy is associated with the onset of several pathological conditions including ageing and age-related neurodegenerative diseases. The nematode Caenorhabditis elegans is a widely used model organism for studying the biology of ageing and neurodegeneration. Here, we describe tools and resources for monitoring mitophagy in C. elegans. We developed two composite, in vivo mitophagy imaging systems based, first, on the Rosella biosensor, which combines a fast-maturing pH-insensitive DsRed fused to a pH-sensitive GFP variant, and second, on a custom, dual-fluorescence reporter system that utilizes a mitochondria-targeted GFP, together with the autophagosomal marker LGG-1/LC3 fused to DsRed. We validated both systems in various cell types and under conditions known to induce mitophagy in the worm. These protocols facilitate non-invasive monitoring of mitophagy in physiologically-relevant contexts.
Keywords
Ageing, autophagosome, autophagy, Caenorhabditis elegans, DsRed, Green fluorescent protein (GFP), lysosomes, fluorescent microscopy, mitochondria, mitophagy, mtRosella
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The authors declare that they have no competing financial interests.
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Subject Areas
Cell biology
Genetics
Biological techniques
Biochemistry
Molecular Biology
Computational biology and bioinformatics
Optics and photonics
Associated Publications
Coordination of mitophagy and mitochondrial biogenesis during ageing in C. elegans
by Konstantinos Palikaras, Eirini Lionaki, and Nektarios Tavernarakis
Nature (11 October, 2015)
An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
Nektarios Tavernarakis
Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology - Hellas, Department of Basic Sciences, Faculty of Medicine, University of Crete, Heraklion 71110, Crete, Greece; Tavernarakis lab
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 12 Oct, 2015
Community comments: 1
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
Mitochondrial dysfunction is a shared hallmark of many human pathologies and ageing. Mitochondrial selective autophagy mediates the removal of damaged or superfluous mitochondria preserving mitochondrial and cellular homeostasis. Deregulation of mitophagy is associated with the onset of several pathological conditions including ageing and age-related neurodegenerative diseases. The nematode Caenorhabditis elegans is a widely used model organism for studying the biology of ageing and neurodegeneration. Here, we describe tools and resources for monitoring mitophagy in C. elegans. We developed two composite, in vivo mitophagy imaging systems based, first, on the Rosella biosensor, which combines a fast-maturing pH-insensitive DsRed fused to a pH-sensitive GFP variant, and second, on a custom, dual-fluorescence reporter system that utilizes a mitochondria-targeted GFP, together with the autophagosomal marker LGG-1/LC3 fused to DsRed. We validated both systems in various cell types and under conditions known to induce mitophagy in the worm. These protocols facilitate non-invasive monitoring of mitophagy in physiologically-relevant contexts.
Figure 1
Figure 2
The authors declare that they have no competing financial interests.
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Associated Publications
Coordination of mitophagy and mitochondrial biogenesis during ageing in C. elegans
by Konstantinos Palikaras, Eirini Lionaki, and Nektarios Tavernarakis
Nature (11 October, 2015)