Reduced Ca2+ uptake by mitochondria in pyruvate dehydrogenase- deficient human diploid fibroblasts

Rodolfo A. Padua, Kyle T. Baron, Bhaskar Thyagarajan, Colin R Campbell, Stanley A Thayer

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12 Scopus citations

Abstract

Physiological and pathological Ca2+ loads are thought to be taken up by mitochondria via a process dependent on aerobic metabolism. We sought to determine whether human diploid fibroblasts from a patient with an inherited defect in pyruvate dehydrogenase (PDH) exhibit a decreased ability to sequester cytosolic Ca2+ into mitochondria. Mobilization of Ca2+ stores with bradykinin (BK) increased the cytosolic Ca2+ concentration ([Ca2+](c)) to comparable levels in control and PDH-deficient fibroblasts. In normal fibroblasts transfected with plasmid DNA encoding mitochondrion- targeted apoaequorin, BK elicited an increase in Ca2+-dependent aequorin luminescence corresponding to an increase in the mitochondrial Ca2+ concentration ([Ca2+](mt)) of 2.0 ± 0.2 μM. The mitochondrial uncoupling agent carbonyl cyanide p-(trifiuoromethoxy)phenylhydrazone blocked the BK- induced [Ca2+](mt) increase, although it did not affect the [Ca2+](c) transient. Basal [Ca2+](c) and [Ca2+](mt) in control and PDH-deficient cells were similar. However, confocal imaging of the potential-sensitive dye JC-1 indicated that the percentage of highly polarized mitochondria was reduced from 30 ± 1% in normal cells to 19 ± 2% in the PDH-deficient fibroblasts. BK-elicited [Ca2+](mt) transients in PDH-deficient cells were reduced to 4% of control, indicating that PDH-deficient mitochondria have a decreased ability to take up cytosolic Ca2+. Thus cells with compromised aerobic metabolism have a reduced capacity to sequester Ca2+.

Original languageEnglish (US)
Pages (from-to)C615-C622
JournalAmerican Journal of Physiology - Cell Physiology
Volume274
Issue number3 43-3
DOIs
StatePublished - Mar 1998

Keywords

  • Aerobic metabolism
  • Apoaequorin gene
  • D-myo-inositol 1,4,5-trisphosphate-sensitive calcium stores
  • Inherited metabolic disorder
  • Intracellular calcium concentration

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