Comparative analysis of CI- and CIV-containing respiratory supercomplexes at single-cell resolution.
brain
in-situ imaging analysis
mitochondria
mitochondrial diseases
mitochondrial dysfunction
mitochondrial respiratory supercomplexes
proximity ligation assay
Journal
Cell reports methods
ISSN: 2667-2375
Titre abrégé: Cell Rep Methods
Pays: United States
ID NLM: 9918227360606676
Informations de publication
Date de publication:
24 05 2021
24 05 2021
Historique:
received:
28
09
2020
revised:
03
02
2021
accepted:
03
03
2021
entrez:
27
4
2022
pubmed:
28
4
2022
medline:
28
4
2022
Statut:
epublish
Résumé
Mitochondria sustain the energy demand of the cell. The composition and functional state of the mitochondrial oxidative phosphorylation system are informative indicators of organelle bioenergetic capacity. Here, we describe a highly sensitive and reproducible method for a single-cell quantification of mitochondrial CI- and CIV-containing respiratory supercomplexes (CI∗CIV-SCs) as an alternative means of assessing mitochondrial respiratory chain integrity. We apply a proximity ligation assay (PLA) and stain CI∗CIV-SCs in fixed human and mouse brains, tumorigenic cells, induced pluripotent stem cells (iPSCs) and iPSC-derived neural precursor cells (NPCs), and neurons. Spatial visualization of CI∗CIV-SCs enables the detection of mitochondrial lesions in various experimental models, including complex tissues undergoing degenerative processes. We report that comparative assessments of CI∗CIV-SCs facilitate the quantitative profiling of even subtle mitochondrial variations by overcoming the confounding effects that mixed cell populations have on other measurements. Together, our PLA-based analysis of CI∗CIV-SCs is a sensitive and complementary technique for detecting cell-type-specific mitochondrial perturbations in fixed materials.
Identifiants
pubmed: 35474694
doi: 10.1016/j.crmeth.2021.100002
pii: S2667-2375(21)00002-3
pmc: PMC9017192
doi:
Substances chimiques
Electron Transport Complex IV
EC 1.9.3.1
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Pagination
100002Informations de copyright
© 2021 The Authors.
Déclaration de conflit d'intérêts
The authors declare no competing interests.
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