Dystrophin expression in muscle stem cells regulates their polarity and asymmetric division

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  Here we found that the pro- portion of YFP 20 Myog + cells per fiber at 72 hsiSCR siDmdYFP Pax7 DAPI Pax7 Myog DAPI during development (i.e., Pax7 − Therefore, we examined satellite stem cells immediately after the first round of cell division in isolated myofibers that were cultured for 42h, and we observed an 80% reduction in the proportion of asymmetric satellite stem cell divisions in myofibers from mdx mice as comparedto those from WT mice (Fig. Notably, PLA also revealed that Changes in the proportions of YFP − satellite stem cells could be explained by their ability to undergo apicobasal asymmetric cell divi-sion to generate one YFP − and one YFP Dmd (siDmd), we confirmed that the low proportion of asymmetric (b) Quantification of Dmd expression (as domain) and Dmd localization in satellitecells from cultured myofibers of WT mice at 0, 12, 24 and 36 h (n = 3 mice; ~50 cells permouse).

2 WT mdx

  DGC-deficient satellite cells display impaired regenerationTo investigate the progression of the myogenic program in dystrophin-deficient satellite cells in vivo, we cardiotoxin-injuredmuscles from WT and mdx mice carrying the Myf5-Cre and R26R-YFP alleles, and we analyzed satellite cells by flow cytometry 3 d after injury (Supplementary Fig. 6a). (c,d) Quantification of the numberof asymmetric divisions relative to the total number of satellite stem cell divisions incultured myofibers from Myf5-Cre:R26R-YFP mice after knockdown of Mark2 (siMark2;n = 5 mice per group; 40 myofibers per mouse) (c) or Pard3 (siPard3; n = 4 mice per group;40 myofibers per mouse) (d), as compared to that from myofibers treated with scramblesiRNA (siSCR).

20 Myf5-Cre:R26R-YFP

  Therefore, to determine the axis of cell division, we immunostained mitotic centrosomes withan antibody specific to the phosphorylated forms of Aurora kinases(p-Aurk) Muscles from WT mice respond to injury through the production ofYFP mdx mice, with a high proportion of myogenic cells that remained in G1 (quadrant Q1 of the FACS plot) (Fig. Immunostaining with antibod- ies to the N terminus, C terminus and rod domain of dystrophinsuggest that full-length dystrophin is expressed in WT satellite cells, whereas shorter isoforms of dystrophin (for example, the Dp71 iso-form that does not interact with Mark2), with transcription start sites after the point mutation in the Dmd allele of mdx mice, may beexpressed in satellite cells from mdx mice.

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  Here we found that the pro- portion of YFP 20 Myog + cells per fiber at 72 hsiSCR siDmdYFP Pax7 DAPI Pax7 Myog DAPI during development (i.e., Pax7 − Therefore, we examined satellite stem cells immediately after the first round of cell division in isolated myofibers that were cultured for 42h, and we observed an 80% reduction in the proportion of asymmetric satellite stem cell divisions in myofibers from mdx mice as comparedto those from WT mice (Fig. Notably, PLA also revealed that Changes in the proportions of YFP − satellite stem cells could be explained by their ability to undergo apicobasal asymmetric cell divi-sion to generate one YFP − and one YFP Dmd (siDmd), we confirmed that the low proportion of asymmetric (b) Quantification of Dmd expression (as domain) and Dmd localization in satellitecells from cultured myofibers of WT mice at 0, 12, 24 and 36 h (n = 3 mice; ~50 cells permouse).

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