Vienne 2017 Anal Biochem

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Vienne JC, Cimetta C, Dubois M, Duburcq T, Favory R, Dessein AF, Fontaine M, Joncquel-Chevalier Curt M, Cuisset JM, Douillard C, Mention-Mulliez K, Dobbelaere D, Vamecq J (2017) A fast method for high resolution oxymetry study of skeletal muscle mitochondrial respiratory chain complexes. Anal Biochem 528:57-62.

» PMID: 28456637

Vienne JC, Cimetta C, Dubois M, Duburcq T, Favory R, Dessein AF, Fontaine M, Joncquel-Chevalier Curt M, Cuisset JM, Douillard C, Mention-Mulliez K, Dobbelaere D, Vamecq J (2017) Anal Biochem

Abstract: High resolution oxymetry study (HROS) of skeletal muscle usually requires 90-120 min preparative phase (dissection, permeabilization and washing). This work reports on the suitability of a rapid muscle preparation which by-passes this long preparation. For a few seconds only, muscle biopsy from pigs is submitted to gentle homogenization at 8000 rotations per minute using an ultra-dispersor apparatus. Subsequent HROS is performed using FCCP instead of ADP, compounds crossing and not plasma membrane, respectively. This simplified procedure compares favorably with classical (permeabilized fibers) HROS in terms of respiratory chain complex activities. Mitochondria from cells undergoing ultradispersion were functionally preserved as attested by relative inefficacy of added cytochrome C (not crossing intact mitochondrial outer membrane) to stimulate mitochondrial respiration. Responsiveness of respiration to ADP (in the absence of FCCP) suggested that these intact mitochondria were outside cells disrupted by ultradispersion or within cells permeated by this procedure.

Copyright © 2017 Elsevier Inc. All rights reserved.

Keywords: electron transfer-pathway (ET-pathway), FCCP, High resolution oxymetry, MIR05 buffer, Muscle needle biopsy, Respiratory chain complex, Saponin-permeabilized fibers, Ultradispersion Bioblast editor: Kandolf G O2k-Network Lab: FR Lille Boutry M, FR Lille Vienne JC


Labels: MiParea: Respiration, Instruments;methods 


Organism: Pig  Tissue;cell: Skeletal muscle  Preparation: Permeabilized tissue, Homogenate 


Coupling state: LEAK, OXPHOS, ET  Pathway: N, S, CIV, ROX  HRR: Oxygraph-2k 

2017-08