Abstract
ABSTRACT: For use in screening for disorders of pyruvate metabolism, a sensitive assay method was developed for measuring the rate of decarboxylation of [1-14C]pyruvate during in vitro culture of skin fibroblasts with dichloroacetate (DCA). The rate of decarboxylation of [1-14C]pyruvate by skin fibroblasts from control subjects increased from 59.6 ± 13.2 to 97.3 ± 12.0 nmol/h/mg protein during in vitro culture in medium supplemented with 10 mM DCA for 3 days. In contrast, the rate hardly increased in cells from four of 20 patients with congenital lactic acidosis of unknown cause during in vitro culture with DCA. On day 3 of culture, the values for the four patients did not overlap those of control cells and so these four patients could be clearly distinguished from control subjects. Measurements of the original activity and the activity of the pyruvate dehydrogenase (PDH) complex after activation with a broad specificity protein phosphatase and DCA suggested that in three of the patients the aberration was a disorder in the mechanism for activation of PDH, including deficiency of PDH phosphatase or a mutation of PDH itself, whereas that in the fourth patient it might be a disorder of the mitochondrial transport system for pyruvate. Thus, measurement of the rate of decarboxylation of [1-14C] pyruvate by skin fibroblasts cultured in medium supplemented with 10 mM DCA for 3 days is a useful method for screening for disorders of pyruvate metabolism in cultured skin fibroblasts.
Similar content being viewed by others
Article PDF
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Naito, E., Kuroda, Y., Takeda, E. et al. Detection of Pyruvate Metabolism Disorders by Culture of Skin Fibroblasts with Dichloroacetate. Pediatr Res 23, 561–564 (1988). https://doi.org/10.1203/00006450-198806000-00005
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1203/00006450-198806000-00005
This article is cited by
-
DNA diagnosis of pyruvate dehydrogenase deficiency in female patients with congenital lactic acidaemia
Journal of Inherited Metabolic Disease (1995)
-
Multiple enzyme defects in mitochondria of a case with congenital lactic acidosis and hyperammonaemia
Journal of Inherited Metabolic Disease (1992)
-
Mutation of E1α gene in a female patient with pyruvate dehydrogenase deficiency due to rapid degradation of E1 protein
Journal of Inherited Metabolic Disease (1992)