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Fig. 5 | BMC Biology

Fig. 5

From: Mutant C. elegans mitofusin leads to selective removal of mtDNA heteroplasmic deletions across generations to maintain fitness

Fig. 5

Mutant Parkin aggravates the adverse effects of the ΔmtDNA;fzo-1. A Schematic representation of experimental setup. The fzo-1 heterozygotes progeny of heteroplasmic pdr-1(mut) hermaphrodites (fzo-1(ht);pdr-1(mut);+/ΔmtDNA) was identified and maintained using self-propagation and single worm genotyping to establish heteroplasmic lines carrying fzo-1(ht);pdr-1(mut);+/ΔmtDNA. Progeny animals (generation 1; G1) were isolated, allowed to lay eggs and their genotypes were determined. The progeny of heteroplasmic pdr-1(mut) hermaphrodites that were fzo-1 mutant (fzo-1(mut);pdr-1(mut);+/ΔmtDNA) or wild type (fzo-1(wt);pdr-1(mut);+/ΔmtDNA) were then monitored over several generations (G2m-G4m and G2wt-G4wt, respectively). B The percent of gravid adults of fzo-1(mut);pdr-1(mut);+/ΔmtDNA mutant progeny across generations (G2m-G4m) and of the stable cross line (> 30 generations, Gm > 30) was determined at the indicated times after egg laying (G2m N = 6, n = 140, G3m N = 5, n = 83, G4m N = 6 n = 89, and Gm > 30 N = 3, n = 116). Data were analyzed using Cox proportional-hazards regression (Additional file 1: Table S2). G2m-G3m were slower to reach adulthood than Gm > 30 (P < 0.001) but not G4m (P = 0.084). C Box plot showing the percent of ΔmtDNA (N > 3 biological repeats) determined in individual animals of the heteroplasmic mutant cross-progeny strains, fzo-1(mut);pdr-1(mut);+/ΔmtDNA (G1(ht) n = 41, G1m-G4m n = 14, 20, 17 and 16, respectively). In the boxplot representation, center line, median; box limits, upper and lower quartiles; whiskers, minimum and maximum; points, data. Data were analyzed using Fractional regression (Additional file 1: Table S3). ΔmtDNA levels of G1m-G4m were significantly lower than those observed in G1(ht), (**) denotes P < 0.001. D Heteroplasmy levels of individual fzo-1(ht);pdr-1(mut);+/ΔmtDNA animals (N > 3 biological repeats) sampled at generations 3–7 (Ght3-Ght7; n = 22) and generations 20–24 (Ght20-Ght24; n = 14). Data were analyzed using the Wilcoxon Mann-Whitney rank sum test (P < 0.001). EH The relative levels of ΔmtDNA (E, G) or +mtDNA (F, H) determined for the gonad and soma (normalized to soma) of a population of EF G2m adults (N = 4) or GH G2wt adults (N = 5). Data were analyzed using the Wilcoxon Mann-Whitney rank sum test, (*) denotes P < 0.05 and (**) denotes P < 0.001. Individual data values are presented in Additional file 2

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