Sperl 1997 Mol Cell Biochem

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Sperl W, Skladal D, Gnaiger E, Wyss M, Mayr U, Hager J, Gellerich FN (1997) High resolution respirometry of premeabilized skeletal muscle fibers in the diagnosis of neuromuscular disorders. Mol Cell Biochem 174:71-8.

» PMID: 9309668

Sperl W, Skladal D, Gnaiger E, Wyss M, Mayr U, Hager J, Gellerich FN (1997) Mol Cell Biochem

Abstract: High resolution respirometry in combination with the skinned fiber technique offers the possibility to study mitochondrial function routinely in small amounts of human muscle. During a period of 2 years, we investigated mitochondrial function in skeletal muscle tissue of 13 patients (average age = 5.8 years). In all of them, an open muscle biopsy was performed for diagnosis of their neuromuscular disorder. Mitochondrial oxidation rates were measured with a highly sensitive respirometer. Multiple substrate-inhibitor titration was applied for investigation of mitochondrial function. About 50 mg fibers were sufficient to obtain maximal respiratory rates for seven different substrates (pyruvate/malate, glutamate/malate, octanoylcarnitine/malate, palmitoylcarnitine /malate, succinate, durochinol and ascorbate/TMPD). Decreased respiration rates with reference to the wet weight of the permeabilized fiber could immediately be detected during the course of measurements. In 4 patients with mitochondrial encephalomyopathy (MEM) the respiration pattern indicated a specific mitochondrial enzyme defect, which was confirmed in every patient by measurements of the individual enzymes (one patient with PDHC deficiency, one with complex I deficiency and two patients with combined complex I and IV deficiency). In the 6 patients with spinal muscular atrophy (SMA) oxidation rates were found to be decreased to 23 ± 5% of controls. The normalized respiration pattern was comparable to that of the controls indicating a decreased content of mitochondria in SMA muscle with normal functional properties. Also in the 3 patients with Duchenne muscular dystrophy (DMD) decreased oxidation rates (42 ± 5%) were detected. In addition a low RCI (1.2) indicated a loose coupling of oxidative phosphorylation in the mitochondria of these patients. It is concluded that investigation of mitochondrial function in saponin skinned muscle fibers using high resolution respirometry in combination with multiple substrate titration offers a valuable tool for evaluation of mitochondrial alterations in muscle biopsies of children suffering from neuromuscular disorders.

Keywords: Skinned muscle fibers, Respirometry, Mitochondrial encephalomyopathy, Spinal muscular atrophy, Muscular dystrophy Duchenne

O2k-Network Lab: AT Innsbruck Gnaiger E, AT Innsbruck Oroboros, AT Salzburg Sperl W, DE Magdeburg Gellerich FN


Labels: MiParea: Respiration, mt-Medicine, Patients 


Organism: Human  Tissue;cell: Skeletal muscle  Preparation: Permeabilized tissue 


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