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Doerrier 2018 MiP2018a
Additional label MitoEAGLE  +
Coupling states LEAK  + , OXPHOS  + , ET  +
Event Oral  +
Has abstract [[Image:MITOEAGLE-logo.jpg|left|100px|link
[[Image:MITOEAGLE-logo.jpg|left|100px|link=|COST Action MitoEAGLE]] In recent years there has been an increasing interest in mitochondrial (mt) research in many physio-pathological conditions. In this context, the evaluation of mt-respiration is crucial for understanding mt-physiology and bioenergetics in health and disease. However, despite of the growing relevance in mt-research, there is no database which provides information about mt-function. [[MitoEAGLE network]] provides a powerful tool for establishing a useful mt-database in different species, tissues, and sample preparations. However, the comparison of reported data on mt-respiration from the scientific literature and inter/intra-laboratory comparisons are very difficult. In permeabilized muscle fibers (pfi) one of the critical issues comparing respiratory data is the quality of the mitochondrial preparations. Therefore, the generation of reference values which allow the evaluation of the skills preparing high-quality pfi, constitutes one of the main aims of the [[WG2]] (muscle) in the [[MitoEAGLE]] project. For this purpose, as a first step, we performed a pilot study collecting mt-respiratory data (pfi from mouse soleus) in different laboratories following the same experimental procedure. In this previous pilot study, we observed heterogeneous results between the groups involved [1]. In the present study, we addressed some potential aspects which could trigger this variability. Mitochondrial respiration was evaluated by [[high-resolution respirometry]] in pfi from soleus muscle from ''N''=15 C57BL6/J male mice using the [[SUIT-008 O2 pfi D014]] protocol (Fig. 1) [2]. Experiments were performed in parallel in the same laboratory by two researchers from different research groups. Our results did not exhibit significant differences on mitochondrial respiration by comparing individual mechanical sample preparations and, the effect of chemicals prepared for this study in both laboratories. On the other hand, we compared the results obtained for each researcher in the former and present study. We detected significant differences in oxygen consumption between both studies. Our findings demonstrate two sources of variability in mitochondrial respiration. First, ADP storage (time and/or temperature) seems to be a critical aspect affecting oxygen fluxes. Wet weight is the other important issue that must be considered and well-explained for generating reliable results in the future. The achievement of the reference values constitutes the starting point for obtaining reliable data in any project, avoiding an under/overestimation that could mask/influence relevant results in a study. This fact will contribute to decrease and maybe avoid non-reproducible data between different laboratories. Finally, this study will benefit the elaboration of a public mt-database in muscle permeabilized tissues.
-database in muscle permeabilized tissues.  +
Has editor [[Plangger M]]  + , [[Kandolf G]]  + , [[Doerrier Velasco CA]]  +
Has title [[Image:DoerrierC.JPG|left|90px|Carolina Anneliese Doerrier]] Inter-laboratory harmonization of protocols for mitochondrial function evaluation in permeabilized muscle fibers.  +
Instrument and method Oxygraph-2k  +
Mammal and model Mouse  +
MiP area Respiration  +
Pathways N  + , S  + , NS  + , ROX  +
Preparation Permeabilized tissue  +
Respiration and regulation ADP  + , Flux control  +
Tissue and cell Skeletal muscle  +
Was published by MiPNetLab AT Innsbruck Oroboros + , ES Barcelona Garcia-Roves PM + , AT Innsbruck Gnaiger E +
Was submitted in year 2018  +
Was submitted to event MiP2018/MitoEAGLE Jurmala LV +
Was written by Doerrier C + , Gama Perez P + , Garcia-Roves PM + , 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.
10:46:53, 17 March 2019  +
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