Serial cryoFIB/SEM Reveals Cytoarchitectural Disruptions in Leigh Syndrome Patient Cells

Structure. 2021 Jan 7;29(1):82-87.e3. doi: 10.1016/j.str.2020.10.003. Epub 2020 Oct 22.

Abstract

The advancement of serial cryoFIB/SEM offers an opportunity to study large volumes of near-native, fully hydrated frozen cells and tissues at voxel sizes of 10 nm and below. We explored this capability for pathologic characterization of vitrified human patient cells by developing and optimizing a serial cryoFIB/SEM volume imaging workflow. We demonstrate profound disruption of subcellular architecture in primary fibroblasts from a Leigh syndrome patient harboring a disease-causing mutation in USMG5 protein responsible for impaired mitochondrial energy production.

Keywords: CryoFIB/SEM; CryoSEM; Leigh syndrome; USMG5; block face; mitochondria disease; volume imaging.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Cells, Cultured
  • Cryoelectron Microscopy / methods
  • Fibroblasts / ultrastructure*
  • Humans
  • Leigh Disease / genetics
  • Leigh Disease / pathology*
  • Mitochondria / ultrastructure
  • Mitochondrial Proton-Translocating ATPases / genetics
  • Mutation
  • Primary Cell Culture / methods

Substances

  • ATP5MK protein, human
  • Mitochondrial Proton-Translocating ATPases