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Mitochondrial calcium response in human transformed lymphoblastoid cells
Authors:Kato Tadafumi  Ishiwata Mizuho  Nagai Takeharu
Affiliation:Laboratory for Molecular Dynamics of Mental Disorders, Brain Science Institute, RIKEN, Hirosawa 2-1, Saitama, 351-0198, Wako, Japan. kato@brain..riken.go.jp
Abstract:Human lymphoblastoid cell line (LCL) transformed by Epstein-Barr Virus (EBV) is a unique cellular model for the study of human diseases. Although pathophysiological significance of mitochondrial calcium regulation is drawing attention, it is not known whether or not mitochondria in LCLs play a role in intracellular calcium signaling. In this study, role of mitochondria of the lymphoblastoid cell line in calcium signaling was examined. Intra-mitochondrial calcium concentration ([Ca2+]m) was successfully measured using dihydro-Rhod-2, revealed by the decrease of fluorescence after application of carbonyl cyanide m-chlorophenylhydrazone (CCCP) and intracellular localization patterns imaged by fluorescent microscope. Platelet activating factor (PAF) concentration-dependently increased cytosolic calcium concentration ([Ca2+]i), while no increase of [Ca2+]m was observed. In contrast, 10 microM thapsigargin increased [Ca2+]i as well as [Ca2+]m. LCLs may be used for the study of possible pathophysiological role of mitochondrial calcium regulation in human diseases.
Keywords:Calcium   Mitochondria   Platelet activating factor   Thapsigargin   Carbonyl cyanide m-chlorophenylhydrazone
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