Lactate Oxidation for the Detection of Mitochondrial Dysfunction in Human Skin Fibroblasts

Ofenstein, John P.; Kiechle, Frederick L.; Dandurand, Diane M.; Belknap, William M.; Moore, Kathleen Healy; Holmes, Richard D.

doi:10.1006/abio.1993.1204

Cited by 11 publications

(9 citation statements)

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“…Indeed, recent findings have revealed that in NALD and RCDP fibroblasts, C 0 2 formation from [14C]-lactate is reduced to 24% and 7%, respectively, compared to that in controls [39]. In parallel, the activity of pyruvate dehydrogenase is reduced by 50%, a similar finding to that in fibroblasts from patients with mitochondria1 defects of unknown aetiology [39].…”

Section: Discussionsupporting

confidence: 68%

“…As octanoate must be converted to acetyl-CoA via mitochondria1 pathways prior to utilization for cholesterol synthesis, it might be expected that part of the labelled acetyl-CoA would be metabolized via the citric acid cycle. Consequently, certain mitochondrial abnormalities, as reported previously in peroxisomal-disease fibroblasts [38,39], should be reflected in the rate and amount of cholesterol synthesized. Indeed, recent findings have revealed that in NALD and RCDP fibroblasts, C 0 2 formation from [14C]lactate is reduced to 24% and 7%, respectively, compared to that in controls [39].…”

Section: Discussionmentioning

confidence: 53%

“…Consequently, certain mitochondrial abnormalities, as reported previously in peroxisomal-disease fibroblasts [38,39], should be reflected in the rate and amount of cholesterol synthesized. Indeed, recent findings have revealed that in NALD and RCDP fibroblasts, C 0 2 formation from [14C]-lactate is reduced to 24% and 7%, respectively, compared to that in controls [39]. In parallel, the activity of pyruvate dehydrogenase is reduced by 50%, a similar finding to that in fibroblasts from patients with mitochondria1 defects of unknown aetiology [39].…”

Section: Discussionmentioning

confidence: 99%

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Cholesterol biosynthesis in dermal fibroblasts from patients with metabolic disorders of peroxisomal origin

Malle

Oettl

Sattler

et al. 1995

Eur J Clin Investigation

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Abstract. As peroxisomes possess some of the integral enzymes for cholesterol biosynthesis, the role of these organelles in cholesterol formation was studied in dermal fibroblasts with three types of peroxisomal defect: group I, characterized by the absence of intact peroxisomes (neonatal adrenoleukodystrophy, cerebrohepatorenal syndrome of Zellweger); group 11, showing impaired activity of a single peroxisomal enzyme (X-linked adrenoleukodystrophy, adrenomyeloneuropathy); and group 111, defective in more than one peroxisomal enzyme (rhizomelic chondrodysplasia punctata). Cells were incubated with three different radioactive precursors, namely [I4C]-octanoate, [I4C]-acetate, and [3H]-mevalonate, and incorporation of these radiolabels into cholesterol was determined. All fibroblasts with peroxisomal defects were able to form cholesterol at concentrations comparable or higher than those in controls dependent on the radioactive substrate. Binding properties (KD and b, , values) of LDL to fibroblasts with peroxisomal defects and downregulation of intracellular cholesterol biosynthesis were similar to those found in fibroblasts from normolipidaemic controls, but different to those observed in LDL-receptor negative fibroblasts. As our studies revealed that cholesterol biosynthesis is not impaired in fibroblasts from patients with metabolic disorders of peroxisomal origin, we conclude that peroxisomes play little or no role in the pathway of cholesterol synthesis beyond mevalonate. In earlier steps of the cholesterol synthesis pathway, peroxisoma1 and mitochondrial defects in parallel may alter cholesterol synthesis indirectly.

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Section: Discussionsupporting

confidence: 68%

Section: Discussionmentioning

confidence: 53%

Section: Discussionmentioning

confidence: 99%

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Cholesterol biosynthesis in dermal fibroblasts from patients with metabolic disorders of peroxisomal origin

Malle

Oettl

Sattler

et al. 1995

Eur J Clin Investigation

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“…b IC 50 (μM ± standard error) in primary enzymatic assay of PDH kinase inhibition (ref ). c EC 50 (μM ± standard error) in cellular assay of increased oxidation of lactate (ref ). d In vivo study in normal Sprague−Dawley rats ( n = 6/group); animals were orally dosed (μmol/kg) after a 24-h fast.…”

mentioning

confidence: 99%

(R)-3,3,3-Trifluoro-2-hydroxy-2-methylpropionamides Are Orally Active Inhibitors of Pyruvate Dehydrogenase Kinase

et al. 1999

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“…PDH activity in fibroblast homogenates was measured by a 14 CO 2 release assay using in situ generation of [1j 14 C] pyruvate from l-[1j 14 C]lactate, modified from Ofenstein et al (1993). The formed 14 CO 2 was collected essentially as described (Sperl et al 1993).…”

Section: Methodsmentioning

confidence: 99%

Four novel PDHA1 mutations in pyruvate dehydrogenase deficiency

Østergaard

Møller

Kalkanoglu-Sivri

et al. 2009

J of Inher Metab Disea

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The pyruvate dehydrogenase (PDH) complex is a mitochondrial multienzyme that catalyses the irreversible oxidative decarboxylation of pyruvate to acetyl-CoA. We report four novel PDHA1 mutations in patients with pyruvate dehydrogenase deficiency. Analysis of PDH activity showed decreased activity in fibroblasts from all four patients, around 16-52% of mean control, similar to what has been found in previous studies. Two of the mutations were missense mutations: c.616G>A (p.Glu206Lys) and c.457A>G (p.Met153Val), one was a 3 bp in-frame deletion: c.429_431delAGG (p.Gly143del), and one was a 65 bp duplication: c.900-6_958dup65. cDNA analysis of the 65 bp duplication showed a small amount of normal transcript in addition to the transcript corresponding to the duplication. The small amount of normal transcript likely explains the survival of the patient, who was a boy. The duplication and one of the missense mutations were associated with decreased amounts of E(1)α And E(1)β protein on western blot analysis, whereas the other two mutations were associated with normal amounts. This study adds four novel mutations to the around 90 reported mutations in PDHA1 (HGMD PDHA1 mutation database). The phenotypes of patients with PDH deficiency have been divided into three groups: a neonatal form with severe lactic acidosis, a form observed only in males and characterized by episodes of ataxia with relapses associated with hyperlactataemia, and an infantile form with hypotonia, lethargy, onset of seizures or dystonia, psychomotor retardation, in some cases Leigh-like lesions and mild to moderate hyperlactataemia. The four patients reported here all belong to the latter group, which is the largest.

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Lactate Oxidation for the Detection of Mitochondrial Dysfunction in Human Skin Fibroblasts

Cited by 11 publications

References 0 publications

Cholesterol biosynthesis in dermal fibroblasts from patients with metabolic disorders of peroxisomal origin

Cholesterol biosynthesis in dermal fibroblasts from patients with metabolic disorders of peroxisomal origin

(R)-3,3,3-Trifluoro-2-hydroxy-2-methylpropionamides Are Orally Active Inhibitors of Pyruvate Dehydrogenase Kinase

Four novel PDHA1 mutations in pyruvate dehydrogenase deficiency

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