Back to article: Impact of F1Fo-ATP-synthase dimer assembly factors on mitochondrial function and organismic aging

FIGURE 3: Impairments of mitochondrial functions affect growth and lifespan in deletion mutants. (A) Comparison of mitochondrial morphology of 3-days and 16-days old wild-type strains with those of ΔPaAtpe and ΔPaAtpg (3 d) by fluorescence microscopy. Mitochondria were stained with MitoTracker® Red and analyzed by confocal fluorescence microscopy. Scale bars: 10 µm. (B) Representative Southern blot analysis of BglII-digested gDNA of wild-type (4-days and 16-days old), ΔPaAtpe and ΔPaAtpg strains (4-days old). The 2500 bp age-specific plDNA (arrow) fragment was detected with a specific plDNA probe. (C) Release of ROS species H2O2 from wild-type (6-days n=3), ΔPaAtpe (6-days n=3) and ΔPaAtpg strains (6-days n = 3) visualized by DAB precipitation. (D) Oxygen consumption (basal respiration) of 6-days old deletion mutant mycelium compared to respiration of 6-days old wild type mycelium (n = 3-4 biological replicates with a total number of 12-16 technical replicates). (E) Relative COX- and AOX-dependent oxygen consumption rate (OCR) of 6-days old wild-type and 6-days old deletion strains mycelium after treatment with KCN (COX-inhibitor) or SHAM (AOX-inhibitor). (F) Relative complex I-dependent OCR of 6-days old wild-type mitochondria compared to mitochondria from 6-days old deletion mutants (n = 4-8 biological replicates with a total number of 15-28 technical replicates). State 4 OCR of mitochondria from wild type was set to 100%. (G) ATP production of 6-days old wild-type and 6-days old deletion mutant mitochondria after removal of samples during state 3 respiration measurements. Samples of three biological replicates and two technical replicates, respectively, were measured three times by a luminescence-based assay. (H) Phenotypic analysis of indicated strains growing on M2 medium for 4 days. (I) Growth rate of wild type (0.58 ± 0.05 cm/d, n = 27), ΔPaAtpe (0.53 ± 0.05 cm/d, n = 25) and ΔPaAtpg (0.53 ± 0.08 cm/d, n =3 3) growing on M2 medium. Growth rate was determined from day 3 to 6 of growth. (J) Lifespan analysis of wild type (mean lifespan = 23.3 d, n = 27), ΔPaATPe (mean lifespan = 15.1 d, n = 25,) and ΔPaATPg (mean lifespan = 13.5 d, n = 33) growing on M2 medium. The error bars represent the standard deviation and the P-values were determined by Student´s t test.

By continuing to use the site, you agree to the use of cookies. more information

The cookie settings on this website are set to "allow cookies" to give you the best browsing experience possible. If you continue to use this website without changing your cookie settings or you click "Accept" below then you are consenting to this. Please refer to our "privacy statement" and our "terms of use" for further information.