Mitochondrial fission and mitophagy are independent mechanisms regulating ischemia/reperfusion injury in primary neurons.
Journal
Cell death & disease
ISSN: 2041-4889
Titre abrégé: Cell Death Dis
Pays: England
ID NLM: 101524092
Informations de publication
Date de publication:
12 05 2021
12 05 2021
Historique:
received:
31
12
2020
accepted:
22
04
2021
revised:
21
04
2021
entrez:
13
5
2021
pubmed:
14
5
2021
medline:
15
10
2021
Statut:
epublish
Résumé
Mitochondrial dynamics and mitophagy are constitutive and complex systems that ensure a healthy mitochondrial network through the segregation and subsequent degradation of damaged mitochondria. Disruption of these systems can lead to mitochondrial dysfunction and has been established as a central mechanism of ischemia/reperfusion (I/R) injury. Emerging evidence suggests that mitochondrial dynamics and mitophagy are integrated systems; however, the role of this relationship in the context of I/R injury remains unclear. To investigate this concept, we utilized primary cortical neurons isolated from the novel dual-reporter mitochondrial quality control knockin mice (C57BL/6-Gt(ROSA)26Sortm1(CAG-mCherry/GFP)Ganl/J) with conditional knockout (KO) of Drp1 to investigate changes in mitochondrial dynamics and mitophagic flux during in vitro I/R injury. Mitochondrial dynamics was quantitatively measured in an unbiased manner using a machine learning mitochondrial morphology classification system, which consisted of four different classifications: network, unbranched, swollen, and punctate. Evaluation of mitochondrial morphology and mitophagic flux in primary neurons exposed to oxygen-glucose deprivation (OGD) and reoxygenation (OGD/R) revealed extensive mitochondrial fragmentation and swelling, together with a significant upregulation in mitophagic flux. Furthermore, the primary morphology of mitochondria undergoing mitophagy was classified as punctate. Colocalization using immunofluorescence as well as western blot analysis revealed that the PINK1/Parkin pathway of mitophagy was activated following OGD/R. Conditional KO of Drp1 prevented mitochondrial fragmentation and swelling following OGD/R but did not alter mitophagic flux. These data provide novel evidence that Drp1 plays a causal role in the progression of I/R injury, but mitophagy does not require Drp1-mediated mitochondrial fission.
Identifiants
pubmed: 33980811
doi: 10.1038/s41419-021-03752-2
pii: 10.1038/s41419-021-03752-2
pmc: PMC8115279
doi:
Substances chimiques
DNM1L protein, human
EC 3.6.5.5
Dynamins
EC 3.6.5.5
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
475Subventions
Organisme : NHLBI NIH HHS
ID : T32 HL007853
Pays : United States
Organisme : NINDS NIH HHS
ID : F31 NS124280
Pays : United States
Organisme : NINDS NIH HHS
ID : R01 NS120322
Pays : United States
Organisme : NINDS NIH HHS
ID : R01 NS076715
Pays : United States
Organisme : NINDS NIH HHS
ID : R42 NS105238
Pays : United States
Organisme : NINDS NIH HHS
ID : R01 NS091242
Pays : United States
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