Hahn 2014 Abstract MiP2014
| Validation of oxygen consumption measurements in muscle and fibroblasts from patients with mitochondrial diseases. |
Link:
MiP2014, Book of Abstracts Open Access
Hahn D, Nuoffer JM (2014)
Event: MiP2014
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; Degenerative Disease and Defect"Mitochondrial Disease; Degenerative Disease and Defect" is not in the list (Cell death, Cryopreservation, Ischemia-reperfusion, Permeability transition, Oxidative stress;RONS, Temperature, Hypoxia, Mitochondrial disease) of allowed values for the "Stress" property. Organism: Human Tissue;cell: Skeletal muscle Preparation: Permeabilized cells Enzyme: Complex I, Complex II; Succinate Dehydrogenase"Complex II; Succinate Dehydrogenase" is not in the list (Adenine nucleotide translocase, Complex I, Complex II;succinate dehydrogenase, Complex III, Complex IV;cytochrome c oxidase, Complex V;ATP synthase, Inner mt-membrane transporter, Marker enzyme, Supercomplex, TCA cycle and matrix dehydrogenases, ...) of allowed values for the "Enzyme" property., Complex IV; Cytochrome c Oxidase"Complex IV; Cytochrome c Oxidase" is not in the list (Adenine nucleotide translocase, Complex I, Complex II;succinate dehydrogenase, Complex III, Complex IV;cytochrome c oxidase, Complex V;ATP synthase, Inner mt-membrane transporter, Marker enzyme, Supercomplex, TCA cycle and matrix dehydrogenases, ...) of allowed values for the "Enzyme" property.
Coupling state: OXPHOS
HRR: Oxygraph-2k
MiP2014: SUIT
Affiliation
Dep Clin Chem, Inselspital, Bern, Switzerland. – dagmar.hahn@insel.ch
Table
| Fibroblasts | Positives | Negatives | Total | Muscle | Positives | Negatives | Total |
|---|---|---|---|---|---|---|---|
| Enzyme activity measurements | 11 | 4 | 15 | 6 | 0 | 6 | |
| Oxygen consumption | 14 | 1 | 15 | 5 | 1 | 6 |
References
- 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.
