Cited by 13Autophagy in Disease and TherapyParkinson's Disease Mechanisms and TreatmentsMitochondrial Function and Pathology

Inhibition of bioenergetics provides novel insights into recruitment of PINK 1‐dependent neuronal mitophagy

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Shin, Yea Seul, Ryall, James G., Britto, Joanne M., Lau, Chew L., Devenish, Rodney J., Nagley, Phillip, Beart, Philip M.·2019·Journal of Neurochemistry

Abstract

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.”

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  1. Scheffler I. E. (2011) Mitochondria. 2nd ed. Wiley.

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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
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

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