Idioma:

Using a quantitative quadruple immunofluorescent assay to diagnose isolated mitochondrial Complex I deficiency

Ahmed, Syeda T ; Alston, Charlotte L ; Hopton, Sila ; He, Langping ; Hargreaves, Iain P ; Falkous, Gavin ; Oláhová, Monika ; McFarland, Robert ; Turnbull, Doug M ; Rocha, Mariana C ; Taylor, Robert W

Scientific reports, 2017-11, Vol.7 (1), p.15676-13, Article 15676 [Revista revisada por pares]

England: Nature Publishing Group

Texto completo disponible

Citas Citado por

Acciones

Título:
Using a quantitative quadruple immunofluorescent assay to diagnose isolated mitochondrial Complex I deficiency
Autor: Ahmed, Syeda T ; Alston, Charlotte L ; Hopton, Sila ; He, Langping ; Hargreaves, Iain P ; Falkous, Gavin ; Oláhová, Monika ; McFarland, Robert ; Turnbull, Doug M ; Rocha, Mariana C ; Taylor, Robert W
Materias: Biopsy ; Cell Nucleus - genetics ; Child ; Child, Preschool ; DNA, Mitochondrial - genetics ; Electron transport ; Electron transport chain ; Electron Transport Complex I - deficiency ; Electron Transport Complex I - genetics ; Female ; Fluorescent Antibody Technique ; Fluoroimmunoassay - methods ; Genetic Heterogeneity ; Genomes ; Humans ; Immunoreactivity ; Male ; Mitochondria - genetics ; Mitochondrial Diseases - diagnosis ; Mitochondrial Diseases - genetics ; Mitochondrial Diseases - pathology ; Mitochondrial DNA ; Muscle, Skeletal - metabolism ; Muscle, Skeletal - pathology ; Mutation ; NADH Dehydrogenase - genetics ; NADH-ubiquinone oxidoreductase ; Oxidative Phosphorylation ; Skeletal muscle ; Spectrophotometry
Es parte de: Scientific reports, 2017-11, Vol.7 (1), p.15676-13, Article 15676
Notas: ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
Descripción: Isolated Complex I (CI) deficiency is the most commonly observed mitochondrial respiratory chain biochemical defect, affecting the largest OXPHOS component. CI is genetically heterogeneous; pathogenic variants affect one of 38 nuclear-encoded subunits, 7 mitochondrial DNA (mtDNA)-encoded subunits or 14 known CI assembly factors. The laboratory diagnosis relies on the spectrophotometric assay of enzyme activity in mitochondrially-enriched tissue homogenates, requiring at least 50 mg skeletal muscle, as there is no reliable histochemical method for assessing CI activity directly in tissue cryosections. We have assessed a validated quadruple immunofluorescent OXPHOS (IHC) assay to detect CI deficiency in the diagnostic setting, using 10 µm transverse muscle sections from 25 patients with genetically-proven pathogenic CI variants. We observed loss of NDUFB8 immunoreactivity in all patients with mutations affecting nuclear-encoding structural subunits and assembly factors, whilst only 3 of the 10 patients with mutations affecting mtDNA-encoded structural subunits showed loss of NDUFB8, confirmed by BN-PAGE analysis of CI assembly and IHC using an alternative, commercially-available CI (NDUFS3) antibody. The IHC assay has clear diagnostic potential to identify patients with a CI defect of Mendelian origins, whilst highlighting the necessity of complete mitochondrial genome sequencing in the diagnostic work-up of patients with suspected mitochondrial disease.
Editor: England: Nature Publishing Group
Idioma: Inglés

Enlaces

View this record in MEDLINE/PubMed

Volver a la lista de resultados

Realización: Logos de Redes Sociales:

Using a quantitative quadruple immunofluorescent assay to diagnose isolated mitochondrial Complex I deficiency

Ahmed, Syeda T ; Alston, Charlotte L ; Hopton, Sila ; He, Langping ; Hargreaves, Iain P ; Falkous, Gavin ; Oláhová, Monika ; McFarland, Robert ; Turnbull, Doug M ; Rocha, Mariana C ; Taylor, Robert W

England: Nature Publishing Group

Buscando en bases de datos remotas, por favor espere