Difference between revisions of "Krylova 2017 MiPschool Obergurgl"
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Complex I (CI) deficiency is one of the most common defects in OXPHOS system and representing more than 30% cases of mitochondrial diseases. The group is characterized by clinical and genetic heterogeneity and comprise several nosological forms. The most prevalent phenotypes for complex I deficiency are LHON and Leigh syndrome. | Complex I (CI) deficiency is one of the most common defects in OXPHOS system and representing more than 30% cases of mitochondrial diseases. The group is characterized by clinical and genetic heterogeneity and comprise several nosological forms. The most prevalent phenotypes for complex I deficiency are LHON and Leigh syndrome. | ||
In this study we analyzed skin fibroblasts from 11 patients with mutations in mtDNA, which cause LHON or Leigh-like phenotypes: m.11778 G>A (N=3), m.3460 A>G (N=2), m.3635 G>A (N=1), m.3308 T>G (N=2), m.3472 T>C (N=1) and 2 patients with unpublished substitutions m.3945 C>A and m.14441T>C. High-resolution respirometry (HRR) on the Oxygraph-2k ( | In this study we analyzed skin fibroblasts from 11 patients with mutations in mtDNA, which cause LHON or Leigh-like phenotypes: m.11778 G>A (N=3), m.3460 A>G (N=2), m.3635 G>A (N=1), m.3308 T>G (N=2), m.3472 T>C (N=1) and 2 patients with unpublished substitutions m.3945 C>A and m.14441T>C. High-resolution respirometry (HRR) on the Oxygraph-2k (Oroboros Instruments, Austria) was performed for complex analysis of mitochondrial respiratory function in intact and permeabilized fibroblasts of patients and healthy controls. | ||
Flux control rations in intact cells R/E, (R-L)/E (p<0,05) were raised compared to the control. Rates R, E, L normalized on the citrate synthase activity (CS) were statistically varied between patients and controls. In permeabilized fibroblasts we observed differences in СII/E, Rot/E, R/CII, CI/CII (p<0,05) between groups. These data highlight the dysfunction of OXPHOS system and particularly CI. Increased CS level and decreased CI/CII ratio indicate compensatory metabolic answer on respiratory chain dysfunction. | Flux control rations in intact cells R/E, (R-L)/E (p<0,05) were raised compared to the control. Rates R, E, L normalized on the citrate synthase activity (CS) were statistically varied between patients and controls. In permeabilized fibroblasts we observed differences in СII/E, Rot/E, R/CII, CI/CII (p<0,05) between groups. These data highlight the dysfunction of OXPHOS system and particularly CI. Increased CS level and decreased CI/CII ratio indicate compensatory metabolic answer on respiratory chain dysfunction. |
Latest revision as of 12:37, 23 January 2019
High-resolution respirometry in diagnostic of mitochondrial complex I deficiency. |
Link: MitoEAGLE
Krylova TD, Tsygankova PG, Itkis YS, Sheremet NL, Nevinitsyna TA, Mikhaylova SV, Zakharova EY (2017)
Event: MiPschool Obergurgl 2017
Complex I (CI) deficiency is one of the most common defects in OXPHOS system and representing more than 30% cases of mitochondrial diseases. The group is characterized by clinical and genetic heterogeneity and comprise several nosological forms. The most prevalent phenotypes for complex I deficiency are LHON and Leigh syndrome.
In this study we analyzed skin fibroblasts from 11 patients with mutations in mtDNA, which cause LHON or Leigh-like phenotypes: m.11778 G>A (N=3), m.3460 A>G (N=2), m.3635 G>A (N=1), m.3308 T>G (N=2), m.3472 T>C (N=1) and 2 patients with unpublished substitutions m.3945 C>A and m.14441T>C. High-resolution respirometry (HRR) on the Oxygraph-2k (Oroboros Instruments, Austria) was performed for complex analysis of mitochondrial respiratory function in intact and permeabilized fibroblasts of patients and healthy controls.
Flux control rations in intact cells R/E, (R-L)/E (p<0,05) were raised compared to the control. Rates R, E, L normalized on the citrate synthase activity (CS) were statistically varied between patients and controls. In permeabilized fibroblasts we observed differences in СII/E, Rot/E, R/CII, CI/CII (p<0,05) between groups. These data highlight the dysfunction of OXPHOS system and particularly CI. Increased CS level and decreased CI/CII ratio indicate compensatory metabolic answer on respiratory chain dysfunction.
Our results show the facilities of HRR in revealing the biochemical abnormalities of complex I in patient’s fibroblasts with LHON and Leigh-like syndrome. We also suggest HRR to be useful method for inspection other mutations causing complex I deficiency.
• Bioblast editor: Kandolf G
• O2k-Network Lab: RU Moscow Itkis YS
Labels: MiParea: Respiration, mtDNA;mt-genetics, mt-Medicine
Tissue;cell: Endothelial;epithelial;mesothelial cell, Fibroblast Preparation: Intact cells, Permeabilized cells
Coupling state: LEAK, ROUTINE, ET
Pathway: N, S
HRR: Oxygraph-2k
Event: B2, Poster
Affiliations
- (1)Krylova TD, (1)Tsygankova PG, (1)Itkis YS, (2)Sheremet NL, (2)Nevinitsyna TA, (3)Mikhaylova SV, (1)Zakharova EY1
- Res Centre Med Gen
- Res Inst Eye Dis
- Rus Child Clin Hosp
- Moscow, Russia. – [email protected]
- (1)Krylova TD, (1)Tsygankova PG, (1)Itkis YS, (2)Sheremet NL, (2)Nevinitsyna TA, (3)Mikhaylova SV, (1)Zakharova EY1