Inhibition of bioenergetics provides novel insights into recruitment of PINK 1‐dependent neuronal mitophagy
Open accessAbstract
Abstract Contributions of damaged mitochondria to neuropathologies have stimulated interest in mitophagy. We investigated triggers of neuronal mitophagy by disruption of mitochondrial energy metabolism in primary neurons. Mitophagy was examined in cultured murine cerebellar granule cells after inhibition of mitochondrial respiratory chain by drugs rotenone, 3‐nitropropionic acid, antimycin A, and potassium cyanide, targeting complexes I, II, III, and IV, respectively. Inhibitor concentrations producing slow cellular demise were determined from analyses of cellular viability, morphology of neuritic damage, plasma membrane permeability, and oxidative phosphorylation. Live cell imaging of dissipation of mitochondrial membrane potential (ΔΨm) by drugs targeting mitochondrial complexes was referenced to complete depolarization by carbonyl cyanide m‐chlorophenyl hydrazone. While inhibition of complexes I, III and IV effected rapid dissipation of ΔΨm, inhibition of complex II using 3‐nitropropionic acid led to minimal depolarization of mitochondria. Nonetheless, all respiratory chain inhibitors triggered mitophagy as indicated by increased aggregation of mitochondrially localized PINK1. Mitophagy was further analyzed using a dual fluorescent protein biosensor reporting mitochondrial relocation to acidic lysosomal environment. Significant acidification of mitochondria was observed in neurons treated with rotenone or 3‐nitropropionic acid, revealing mitophagy at distal processes. Neurons treated with antimycin A or cyanide failed to show mitochondrial acidification. Minor dissipation of ΔΨm by 3‐nitropropionic acid coupled with vigorous triggering of mitophagy suggested depolarization of mitochondria is not a necessary condition to trigger mitophagy. Moreover, weak elicitation of mitophagy by antimycin A, subsequent to loss of ΔΨm, suggested that mitochondrial depolarization is not a sufficient condition for triggering robust neuronal mitophagy. Our findings provide new insight into complexities of mitophagic clearance of neuronal mitochondria. image
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Mild mitochondrial uncoupling induced by 3-nitropropionic acid is sufficient to trigger massive mitophagy in mouse neurons.
“A mild mitochondrial uncoupling as induced by 3-nitropropionic acid (–30% in TMRM staining, a fluorescent probe used to assess mitochondrial potential membrane) is sufficient to trigger massive mitophagy in mouse neurons.”
From Mitochondrial Uncoupling: A Key Controller of Biological Processes in Physiology and Diseases (2019) · cites this paper as [112]
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- Authors
- Shin, Y. S., Ryall, J. G., Britto, J. M., Lau, C. L., Devenish, R. J., Nagley, P., & Beart, P. M.
- Affiliations
- The University of Melbourne (AU)
- Florey Institute of Neuroscience and Mental Health (AU)
- Monash University (AU)
- Corresponding author
- Beart, Philip M.
- Venue
- Journal of Neurochemistry
- Publisher
- Wiley
- Published
- 2019-02-12
- Volume
- 149
- Issue
- 2
- Pages
- 269-283
- ISSN
- 0022-3042 (ISSN-L)
- 1471-4159
- OpenAlex
- W2915015587
- Cited by
- 13
- References
- 74
- Citations by year
- 2026: 1 · 2025: 1 · 2023: 2 · 2022: 2 · 2021: 4 · 2020: 2 · 2019: 1
- FWCI
- 1.71
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Journal of Neurochemistry (publishedVersion)
- PubMed (publishedVersion)
- Minerva Access (University of Melbourne) (submittedVersion)
- Funders
- National Health and Medical Research Council (APP1020401, APP509217)
- Topics
- Health Sciences › Medicine › Epidemiology › Autophagy in Disease and Therapy
- Health Sciences › Medicine › Neurology › Parkinson's Disease Mechanisms and Treatments
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Mitochondrial Function and Pathology
- Keywords
- PINK1, mitochondrial respiratory chain inhibition, mitochondrial depolarization, mitochondrial membrane potential, 3-nitropropionic acid, rotenone, antimycin A, potassium cyanide, cerebellar granule cells, mitochondrial bioenergetics
- MeSH terms
- PTEN-Induced Putative Kinase, Animals, Cells, Cultured, Energy Metabolism, Neurons, Protein Kinases, Mice, Membrane Potential, Mitochondrial, Mitophagy
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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