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Doerrier 2018 MiP2018b
Additional label MitoEAGLE  +
Coupling states OXPHOS  +
Event Oral  +
Has abstract [[Image:MITOEAGLE-logo.jpg|left|100px|link
[[Image:MITOEAGLE-logo.jpg|left|100px|link=|COST Action MitoEAGLE]] In recent years, research of mitochondrial (mt) function received an increasing interest in the context of many pathophysiological conditions. Identification of metabolic reprogramming and specific defects in mt-pathways is critical for diagnosis and selection of targeted treatments. Fatty acid oxidation (FAO, F-pathway) plays an essential role in many pathologies (e.g. insulin resistance, heart failure, ischemia-reperfusion, cancer) and must therefore be studied carefully. Here we show that respirometric measurements can significantly overestimate F-pathway capacities due to activation of anaplerotic pathways (e.g. mt-malic enzyme) [1,2]. F-pathway respiration requires the combined use of fatty acid(s) (FA) and malate to avoid acetyl-CoA accumulation which inhibits mt-respiration. In the presence of enzymes supporting malate- and pyruvate-linked anaplerotic pathways the widely used malate concentration of 2 mM supports NADH-linked (N-) respiration in addition to the F-pathway, resulting in overestimation of F-OXPHOS capacity. We developed the substrate-uncoupler-inhibitor titration (SUIT) protocol RP2 for reliable FAO evaluation by high-resolution respirometry (HRR). By comparison, we quantified the degree of FAO overestimation in several mouse and human mt-preparations. In conclusion, HRR allows the accurate study of mt-pathways, including F-pathway capacity, which is required for comprehensive OXPHOS analysis in diagnostic studies of mt-function.
ysis in diagnostic studies of mt-function.  +
Has editor [[Plangger M]]  +
Has title [[Image:DoerrierC.JPG|left|90px|Carolina Doerrier]] Evaluation of anaplerotic pathways to avoid artefacts in respirometric measurement of fatty acid oxidation.  +
Instrument and method Oxygraph-2k  +
Mammal and model Human  + , Mouse  +
MiP area Instruments;methods  + , Respiration  +
Pathways F  + , N  +
Preparation Homogenate  + , Isolated mitochondria  + , Permeabilized cells  +
Respiration and regulation Fatty acid  +
Stress Cryopreservation  +
Tissue and cell Blood cells  + , HEK  + , Heart  + , Nervous system  + , Platelet  +
Was published by MiPNetLab AT Innsbruck Oroboros + , AT Innsbruck Gnaiger E +
Was submitted in year 2018  +
Was submitted to event MiP2018/MitoEAGLE Jurmala LV +
Was written by Doerrier C + , Garcia-Souza LF + , Sumbalova Z + , Krumschnabel G + , Gnaiger E +
Categories Abstracts
Modification date
"Modification date" is a predefined property that corresponds to the date of the last modification of a subject and is provided by Semantic MediaWiki.
19:56:19, 17 March 2019  +
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