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 withunprecedented detail. Our goal is to test the feasibility of cryo-ET to visualize mitochondrial structural changes in patientcells. We hypothesize in situ cryo-ET will resolve structural changes in the individual mitochondrial respiratory complexesand organization of these complexes into higher-order supercomplexes. We have now obtained primary fibroblast cells frompatients with distinct mitochondrial complex I mutations including ND6, ACAD9, and NDUFV1 subunits and optimizedgrowing these mutant cells on EM grids. We characterized effects of these mutations on mitochondrial morphology anddynamics in living patient cells. In situ cryo-ET revealed disruptions of mitochondrial inner membranes and cristamorphology distinct to each mutant. Our results suggest these respiratory complex subunits are key regulators of overallmitochondrial structure and function and their disruption directly alters their function to produce profound human disease.
Document Details
- Document Type
- Technical Report
- Publication Date
- Mar 01, 2022
- Accession Number
- AD1172662
Entities
People
- Zachary Freyberg
Organizations
- University of Pittsburgh