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Restoring Systemic GDF11 Levels Reverses Age-Related Dysfunction in Mouse Skeletal Muscle

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Science  09 May 2014:
Vol. 344, Issue 6184, pp. 649-652
DOI: 10.1126/science.1251152

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  1. Fig. 1 Rejuvenation of muscle stem cells by heterochronic parabiosis.

    (A) Frequency of clone-sorted satellite cells from isochronic (Iso) or heterochronic (Het) mice forming colonies after 5 days in culture. All colonies showed characteristic morphology of muscle lineage cells. (B) DNA damage in freshly sorted satellite cells assessed by single-cell gel electrophoresis under alkaline conditions. Damage was quantified using a visual scoring metric (25) (key at top) and represented by color coding: no damage, green; moderate damage, orange; maximal damage, red. (C) Representative images (confocal z stacks) of freshly sorted satellite cells stained with 4′,6-diamidino-2-phenylindole (DAPI) (blue) and antibody to pH2AX (green). Data are quantified in (D). All graphs represent mean ± SD, with P values calculated by Mann-Whitney analysis. n, number of mice used for each analysis. Scale bar, 10 μm.

  2. Fig. 2

    Rejuvenation of muscle stem cells by rGDF11 supplementation. (A and B) Frequency (A) and myogenic colony formation (B) of satellite cells from vehicle- or rGDF11-treated mice. (C) Quantification of DNA damage assays using freshly sorted satellite cells from vehicle- or rGDF11-treated mice, scored as in Fig. 1B. (D and E) Hematoxylin and eosin staining (D) and frequency distribution of myofiber size (E) in regenerating TA muscles 7 days after cryoinjury in vehicle- or rGDF11-treated young and aged mice. Scale bar, 100 μm. (F) Representative images of transverse cryosections of TA muscles 2 weeks after transplantation. (G) Quantification of transplant data as maximal number of GFP+ myofibers found in each engrafted muscle. Graphs represent mean ± SD [in (A) to (C) and (G)]. P values were calculated by Mann-Whitney analysis [(A) to (C)], Wilcoxon Exact analysis (E), or Student’s t test (G). n, number of mice used for each analysis.

  3. Fig. 3

    Improved muscle physiology and physical function after rGDF11 supplementation. (A) Electron micrographs of transverse sections of TA muscle from vehicle- or rGDF11-treated aged mice (representative of n = 4 mice per group). Arrows indicate swollen mitochondria. (B and C) Western blot of PGC-1α (B) and LC3 forms I and II (C) in TA muscle extracts from cardiotoxin-injured or uninjured vehicle- or rGDF11-treated aged mice. Three animals are shown for each experimental group. Densitometric quantification of Western data are provided below each blot, normalized to glyceraldehyde phosphate dehydrogenase (GAPDH) (B) or Actin (C). (D and E) Scatter plots of exercise endurance [(D), maximum treadmill runtime in a 90-min window] or forelimb grip strength (E) of vehicle- or rGDF11-treated aged mice. Grip strength is plotted as maximum force (Newton, N) exerted in triplicate trials. The red line represents the maximum grip strength of 33- to 39-week-old young male mice. Data are presented for individual mice (black symbols) overlaid with mean ± SD (orange lines). P values were calculated by Mann-Whitney analysis. n, number of mice used for each analysis.

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