Hahn 2014 Abstract MiP2014
|Validation of oxygen consumption measurements in muscle and fibroblasts from patients with mitochondrial diseases.|
Diagnosis of mitochondrial disorders is mainly based on the analysis of OXPHOS complexes in muscle biopsies. However, normal enzyme activities do not rule out the presence of a mitochondrial disorder. Therefore, analysis of the integrated mitochondrial energy generating system by oxygen consumption is frequently used.
We compared the diagnostic value of respiratory versus enzymatic OXPHOS analysis in fibroblasts and muscle biopsies from patients with a genetically confirmed mitochondrial disease. A standardized substrate-uncoupler-inhibitor-titration (SUIT) protocol [1,2] was used for measuring respiration of permeabilized fibroblasts and single muscle fibers. OXPHOS enzyme activities were determined spectrophotometrically according to standard protocols. Only the combination of both measurements enables us to identify a mitochondrial disorder in all of the present patients (Table).
Moreover, specific flux control ratios showed higher diagnostic sensitivity than complex specific O2 fluxes. The established SUIT protocol is an important tool in the diagnostic process. Therefore, we recommend oxygen consumption measurements in addition to enzymatic OXPHOS analysis, to increase the number of identified patients.
• O2k-Network Lab: CH Bern Nuoffer JM
Labels: MiParea: Respiration, mt-Medicine, Patients
Stress:Mitochondrial disease Organism: Human Tissue;cell: Skeletal muscle, Fibroblast Preparation: Permeabilized cells Enzyme: Complex I, Complex II;succinate dehydrogenase, Complex IV;cytochrome c oxidase
Coupling state: OXPHOS
HRR: Oxygraph-2k Event: C1, Poster MiP2014: SUIT
Dep Clin Chem, Inselspital, Bern, Switzerland. – [email protected]
|Enzyme activity measurements||11||4||15||6||0||6|
- Hirsch A, Hahn D, Kempna P, Hofer G, Mullis P, Nuoffer JM, Flück CE (2012) Role of AMP-activated protein kinase on steroid hormone biosynthesis in adrenal NCI-H295R cells. PLoS ONE 7: e30956.
- Jackson CB, Nuoffer JM, Hahn D, Prokisch H, Haberberger B, Gautsch M, Häberli A, Gallati S, Schaller A (2014) Mutations in SDHD lead to autosomal recessive encephalomyopathy and isolated mitochondrial Complex II deficiency. J Med Genet 51: 170-5.