A New In Situ Cryo-Electron Microscopy Approach to Directly Visualize Mutations in Mitochondrial Disease
Abstract
Mitochondrial dysfunction is associated with several chronic, deployment-associated conditions in Veterans. In situ cryoelectron tomography (cryo-ET) imaging is a new 3D imaging approach that we have used to study patient cells with unprecedented detail. Our goal is to test the feasibility of cryo-ET to visualize mitochondrial structural changes in patient cells. We hypothesize in situ cryo-ET will resolve structural changes in the individual mitochondrial respiratory complexes and organization of these complexes into higher-order super complexes. We have now obtained primary fibroblast cells from patients with distinct mitochondrial complex I mutations including ND6, ACAD9, and NDUFV1 subunits and optimized growing these mutant cells on EM grids. We characterized effects of these mutations on mitochondrial morphology and dynamics in living patient cells. In situ cryo-ET revealed disruptions of mitochondrial inner membranes and crista morphology distinct to each mutant. Our results suggest these respiratory complex subunits are key regulators of overall mitochondrial structure and function and their disruption directly alters their function to produce profound human disease.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jun 01, 2023
- Accession Number
- AD1209364
Entities
People
- Zachary Freyberg
Organizations
- University of Pittsburgh