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Updated: Jun 19, 2026

Visualization of ATP Synthase Dimers in Mitochondria by Electron Cryo-tomography
10:39

Visualization of ATP Synthase Dimers in Mitochondria by Electron Cryo-tomography

Published on: September 14, 2014

In situ cryo-ET visualization of mitochondrial depolarization and mitophagic engulfment

Kevin Rose1,2,3, Eric Herrmann1,2,3, Eve Kakudji1,2,3

  • 1Aligning Science Across Parkinson's (ASAP) Collaborative Research Network, Chevy Chase, MD, USA.

Biorxiv : the Preprint Server for Biology
|April 8, 2025
PubMed

Insights

Mitochondrial depolarization triggers structural changes, including fragmentation and altered ATP synthase positioning, observed via cryo-electron tomography. These findings illuminate cellular responses relevant to Parkinson's disease and mitophagy.

Area of Science:

  • Cell Biology
  • Neuroscience
  • Biochemistry

Background:

  • Mitochondrial dysfunction and impaired quality control are linked to Parkinson's disease, particularly mutations in PINK1 and PRKN.
  • Understanding the structural consequences of mitochondrial depolarization is crucial for elucidating disease mechanisms.

Purpose of the Study:

  • To visualize and characterize the nano-structural changes in mitochondria following depolarization using in situ cryo-electron tomography (cryo-ET).
  • To investigate the association of phagophores and lipid transporters with depolarized mitochondria during mitophagy.

Main Methods:

  • Utilized in situ cryo-electron tomography (cryo-ET) on Parkin-expressing U2OS cells treated with oligomycin and antimycin A (OA).
  • Performed cryo-FIB milling to prepare samples for high-resolution imaging of mitochondrial ultrastructure.
  • Employed sub-tomogram averaging to determine the structure of the prohibitin complex in situ.

Main Results:

  • Observed mitochondrial fragmentation, loss of calcium phosphate crystals, and altered cristae structure (narrowed intermembrane gap, reduced volume).
  • Detected phagophores associated with mitochondrial fragments and potential ATG2A-linked bridge-like densities.
  • Found re-localization of ATP synthases to the inner boundary membrane and identified open/closed conformations of the prohibitin complex, with enrichment of the closed form after OA treatment.

Conclusions:

  • Provided high-resolution, in situ snapshots of mitochondrial structural changes induced by depolarization.
  • Established a structural baseline for studying Parkin-dependent mitophagy and its role in Parkinson's disease.
  • Revealed novel insights into the molecular machinery involved in mitophagic processes.

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