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TitleSnapshots of mitochondrial fission imaged by cryo-scanning transmission electron tomography.
Journal, issue, pagesJ Cell Sci, Vol. 138, Issue 9, Year 2025
Publish dateMay 1, 2025
AuthorsPeter Kirchweger / Sharon Grayer Wolf / Neta Varsano / Tali Dadosh / Guenter P Resch / Michael Elbaum /
PubMed AbstractMitochondria undergo constant remodeling via fission, fusion, extension and degradation. Fission, in particular, depends on the accumulation of mitochondrial fission factor (MFF) and subsequent ...Mitochondria undergo constant remodeling via fission, fusion, extension and degradation. Fission, in particular, depends on the accumulation of mitochondrial fission factor (MFF) and subsequent recruitment of the dynamin-related protein DRP1 (also known as DNM1L). We used cryo-scanning transmission electron tomography (cryo-STET) to investigate mitochondrial morphologies in MFF mutant (MFF-/-) mouse embryonic fibroblast (MEF) cells in ATP-depleting conditions that normally induce fission. The capability of cryo-STET to image through the cytoplasmic volume to a depth of 1 µm facilitated visualization of intact mitochondria and their surroundings. We imaged changes in mitochondrial morphology and cristae structure, as well as contacts with the endoplasmic reticulum (ER), degradative organelles and the cytoskeleton at stalled fission sites. We found disruption of the outer mitochondrial membrane at contact sites with the ER and degradative organelles at sites of mitophagy. We identified fission sites where the inner mitochondrial membrane is already separated while the outer membrane is still continuous. Although MFF is a general fission factor, these observations demonstrate that mitochondrial fission can proceed to the final stage in its absence. The use of cryo-STET allays concerns about the loss of structures due to sample thinning required for tomography using cryo-transmission electron microscopy.
External linksJ Cell Sci / PubMed:40365741
MethodsEM (tomography)
Structure data

EMDB-51759: Dual-Axis cryo-STEM tomogram of a Mitochondrion from MFF-/- MEFs
Method: EM (tomography)

EMDB-51763: CSTET Tomogram of a Mitochondrion from MFF-/- MEFs under fission inducing conditions
Method: EM (tomography)

EMDB-51764: Dual-axis CSTET tomogram of mitochondria from a MFF-/- MEF cell, under fission-inducing conditions
Method: EM (tomography)

EMDB-53197: Dual-axis CSTET tomogram of mitochondria from MFF-/- U2-OS cells under fission-inducing conditions
Method: EM (tomography)

Source
  • Mus musculus (house mouse)
  • Homo sapiens (human)

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