Microscale oxygraphy reveals OXPHOS impairment in MRC mutant cells.

TitleMicroscale oxygraphy reveals OXPHOS impairment in MRC mutant cells.
Publication TypeJournal Article
Year of Publication2012
AuthorsInvernizzi, F, D'Amato, I, Jensen, PB, Ravaglia, S, Zeviani, M, Tiranti, V
JournalMitochondrion
Volume12
Issue2
Pagination328-35
Date Published2012 Mar
ISSN1872-8278
KeywordsClinical Laboratory Techniques, Humans, Mitochondria, Mitochondrial Diseases, Oxidative Phosphorylation, Sensitivity and Specificity, Time Factors
Abstract

Given the complexity of the respiratory chain structure, assembly and regulation, the diagnostic workout for the identification of defects of oxidative phosphorylation (OXPHOS) is a major challenge. Spectrophotometric assays, that measure the activity of individual respiratory complexes in tissue and cell homogenates or isolated mitochondria, are highly specific, but their utilization is limited by the availability of sufficient biological material and intrinsic sensitivity. A further limitation is tissue specificity, which usually determines attenuation, or disappearance, in cultured fibroblasts, of defects detected in muscle or liver. We used numerous fibroblast cell lines derived from patients with OXPHOS deficiencies to set up experimental protocols required for the direct readout of cellular respiration using the Seahorse XF96 apparatus, which measures oxygen consumption rate (OCR) and extra-cellular acidification rate (ECAR) in 96 well plates. Results demonstrate that first level screening based on microscale oxygraphy is more sensitive, cheaper and rapid than spectrophotometry for the biochemical evaluation of cells from patients with suspected mitochondrial disorders.

DOI10.1016/j.mito.2012.01.001
Alternate JournalMitochondrion
Citation Key10.1016/j.mito.2012.01.001
PubMed ID22310368
PubMed Central IDPMC3314980
Grant ListGGP11088 / / Telethon / Italy
GPP10005 / / Telethon / Italy