Abstract
Satellite cells are the resident stem cells of adult skeletal muscle, supplying myonuclei for homoeostasis, hypertrophy and repair. In this study, we have examined the role of bone morphogenetic protein (BMP) signalling in regulating satellite cell function. Activated satellite cells expressed BMP receptor type 1A (BMPR-1A/Alk-3) and contained phosphorylated Smad proteins, indicating that BMP signalling is operating during proliferation. Indeed, exogenous BMP4 stimulated satellite cell division and inhibited myogenic differentiation. Conversely, interfering with the interactions between BMPs and their receptors by the addition of either the BMP antagonist Noggin or soluble BMPR-1A fragments, induced precocious differentiation. Similarly, blockade of BMP signalling by siRNA-mediated knockdown of BMPR-1A, disruption of the intracellular pathway by either Smad5 or Smad4 knockdown or inhibition of Smad1/5/8 phosphorylation with Dorsomorphin, also caused premature myogenic differentiation. BMP signalling acted to inhibit the upregulation of genes associated with differentiation, in part, through regulating Id1. As satellite cells differentiated, Noggin levels increased to antagonise BMP signalling, since Noggin knockdown enhanced proliferation and impeded myoblast fusion into large multinucleated myotubes. Finally, interference of normal BMP signalling after muscle damage in vivo perturbed the regenerative process, and resulted in smaller regenerated myofibres. In conclusion, BMP signalling operates during routine satellite cell function to help coordinate the balance between proliferation and differentiation, before Noggin is activated to antagonise BMPs and facilitate terminal differentiation.
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Abbreviations
- BMP:
-
bone morphogenetic protein
- BMPR-1A:
-
bone morphogenetic protein receptor type 1A/Alk-3
- sBMPR-1Af:
-
soluble bone morphogenetic protein receptor 1A fragments
- caBMPR-1A:
-
constitutively active bone morphogenetic protein receptor type 1A
- pSmad1/5/8:
-
phosphorylated Smad1, Smad5 and Smad8
- Id:
-
inhibitor of differentiation/DNA-binding protein
- FOP:
-
Fibrodysplasia ossificans progressiva
- EDL:
-
extensor digitorum longus
- MyHC:
-
myosin heavy chain
- CKM:
-
Muscle creatine kinase
- DAPI:
-
4,6-diamidino-2-phenylindole
- T0:
-
freshly isolated
- T48:
-
cultured for 48 h
- T72:
-
cultured for 72 h
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Acknowledgements
We thank Dr. Stephen J. Tapscott for providing the CKM-LUCpCS2 reporter plasmid; Dr. Robert Benezra for generously sharing pcDNA3-mId1 construct; Dr. Naohiro Hashimoto for helpful discussions and Paul Knopp for much help. We gratefully acknowledge colleagues who shared antibodies, including through the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the University of Iowa. Y.O. was funded by the Muscular Dystrophy Campaign (RA3/737); F.C. is sponsored by the Association of International Cancer Research (07-0151); J.E.M. is backed by a Wellcome Trust university award. The laboratory of P.S.Z. is also supported by The Wellcome Trust, The Medical Research Council, the MYORES Network of Excellence (contract 511978) from the European Commission 6th Framework Programme and the collaborative Project OPTISTEM (Grant Agreement number: 223098) from the European Commission 7th Framework Programme.
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Ono, Y., Calhabeu, F., Morgan, J. et al. BMP signalling permits population expansion by preventing premature myogenic differentiation in muscle satellite cells. Cell Death Differ 18, 222–234 (2011). https://doi.org/10.1038/cdd.2010.95
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DOI: https://doi.org/10.1038/cdd.2010.95
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