posted on 2011-12-31, 01:56authored byDeborah Pye, Dimitra S. Kyriakouli, Geoffrey A. Taylor, Riem Johnson, Matthias Elstner, Brigitte Meunier, Zofia M. A. Chrzanowska-Lightowlers, Robert W. Taylor, Douglass M. Turnbull, Robert N. Lightowlers
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Taken from "Production of mitochondrial cybrids containing naturally occurring pathogenic mtDNA variants"
() Whole cell suspensions of mitochondrial cybrids 1.12 and 1.14 along with 143B. ρ and parental cells were dithionite reduced and scanned at room temperature. Distinct peaks can be seen corresponding to cytochrome + (c), (b) and (a) for the 143B parental line. There is little evidence of cytochrome in either of the clones or the ρ line a decreased level of complex IV. Cytochrome depletion is also noted in 1.12, consistent with defective mitochondrial translation. () Cells (143B parental, clone 1.14, clone 1.12) were suspended in incubation media and assessed for respiration as detailed in Materials and Methods. Clone 1.12 exhibits a major decrease in endogenous respiration rate when compared to the control 143B parental cells. All lines showed a stimulation of activity on uncoupling of the respiratory chain by dinitrophenol (UNC). Again, the respiration rate of 1.12 with all respiratory substrates was substantially reduced, consistent with a defect in mitochondrial translation. Clone 1.14 showed a decreased rate of respiration when compared to 143B parentals, but this depletion was not marked, suggesting that complex IV does not have a major control strength for respiration in these cells.