New method to assess mitophagy flux by flow cytometry
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Mitochondrial autophagy, also known as mitophagy, is an autophagosome-based mitochondrial degradation process that eliminates unwanted or damaged mitochondria after cell stress. Most studies dealing with mitophagy rely on the analysis by fluorescence microscopy of mitochondrial-autophagosome colocalization. However, given the fundamental role of mitophagy in the physiology and pathology of organisms, there is an urgent need for novel quantitative methods with which to study this process. Here, we describe a flow cytometry-based approach to determine mitophagy by using MitoTracker Deep Red, a widely used mitochondria-selective probe. Used in combination with selective inhibitors it may allow for the determination of mitophagy flux. Here, we test the validity of the use of this method in cell lines and in primary cell and tissue cultures.
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MitoTracker Deep Red fluorescence was significantly greater compared to MitoTracker Red in APOE4 astrocytes.
“their fluorescence of MitoTracker Deep Red, which is less sensitive to MMP21,24,25, was significantly greater compared to MitoTracker Red (Fig. 5b)”
From Altered mitochondrial dynamics and function in APOE4-expressing astrocytes (2020) · cites this paper as [24]
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Altered mitochondrial dynamics and function in APOE4-expressing astrocytes
Schmukler, E., Solomon, S., Simonovitch, S., Goldshmit, Y., Wolfson, E., Michaelson, D. M., & Pinkas-Kramarski, R. (2020) · Cell Death & Disease · doi:10.1038/s41419-020-02776-4 · Cited by 125 · see citing works
- Authors
- Mauro-Lizcano, M., Esteban-Martínez, L., Seco, E., Serrano-Puebla, A., Garcia-Ledo, L., Figueiredo-Pereira, C., Vieira, H. L. A., & Boya, P.
- Affiliations
- Consejo Superior de Investigaciones Científicas (ES)
- Universidade Nova de Lisboa (PT)
- Instituto de Biologia Experimental e Tecnológica (PT)
- Corresponding author
- Boya, Patricia
- Venue
- Autophagy
- Publisher
- Informa UK Limited
- Published
- 2015-05-04
- Volume
- 11
- Issue
- 5
- Pages
- 833-843
- ISSN
- 1554-8627 (ISSN-L)
- 1554-8635
- PMC
- PMC4509449
- OpenAlex
- W1642988509
- Cited by
- 140
- References
- 10
- Citations by year
- 2026: 9 · 2025: 9 · 2024: 11 · 2023: 11 · 2022: 15 · 2021: 14 · 2020: 17 · 2019: 9 · 2018: 18 · 2017: 12 · 2016: 13 · 2015: 1
- FWCI
- 10.55
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Autophagy (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Portuguese National Funding Agency for Science, Research and Technology (RCAAP Project by FCT) (submittedVersion, other-oa)
- Figshare (submittedVersion, cc-by)
- Figshare (cc-by)
- Topics
- Health Sciences › Medicine › Epidemiology › Autophagy in Disease and Therapy
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Physiology › Calcium signaling and nucleotide metabolism
- Keywords
- mitophagy, flow cytometry assay, mitochondrial degradation, primary cell culture, tissue culture
- MeSH terms
- Amino Acids, Animals, Carbonyl Cyanide m-Chlorophenyl Hydrazone, Disease Models, Animal, Embryo, Mammalian, Fibroblasts, Flavonoids, Flow Cytometry, HeLa Cells, Humans, Lysosomes, Mice, Inbred C57BL, Mitochondria, Niacinamide, Retina, Retinitis Pigmentosa, Down-Regulation, Flavonols, Mitophagy
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