Decorin modulates collagen I-stimulated, but not fibronectin-stimulated, migration of C2C12 myoblasts.
Extracellular matrix factors, specifically fibronectin and collagen I, are essential for structural support during muscle regeneration. Decorin has been identified as an anti-fibrotic agent with binding sites located on both fibronectin and collagen I. Upon injury, activated myoblasts are required to migrate through the extracellular matrix factors deposited by the myofibroblasts to facilitate skeletal muscle regeneration. In this study we looked at the effects decorin on fibronectin- and collagen I-stimulated myoblast migration. Dose response studies demonstrated 10 μg/ml, 5 μg/ml and 25 μg/ml as the optimal stimulatory concentrations of decorin (1.2 fold increase), fibronectin (3.5 fold increase) and collagen I (2.4 fold increase), when compared with control respectively. A synergistic effect was identified when decorin and collagen I were added in combination; this effect was not evident when decorin was added with fibronectin. The effects of these factors on the ROCK signalling pathway were also analyzed. ROCK-2 was identified as the key Rho-activated kinase isoform involved in migration, due to its higher expression levels and localisation to focal points within migrating C2C12 myoblasts. Decorin and collagen I in combination stimulated an increase in the number of ROCK-2 localized focal points when compared with control, decorin and collagen I added separately. Fibronectin did not show any increase in ROCK-2 focal points when compared with control. These results show for the first time that decorin can modify collagen I-stimulated, but not fibronectin-stimulated myoblast migration in vitro. Furthermore, the synergistic, rather than additive, effect observed suggests a direct modification of collagen I signalling by decorin mediated, at least in part, by ROCK-2 rather than ROCK-1.
Department of Biochemistry, School of Biochemistry, Genetics and Microbiology, University of KwaZulu-Natal, Scottsville, South Africa.
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Goetsch
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Kallmeyer
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Niesler
D000818:Animals; D002417:Cattle; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects* / Q000502:physiology; D024042:Collagen Type I / Q000494:pharmacology*; D058575:Decorin / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D005353:Fibronectins / Q000494:pharmacology*; D022001:Focal Adhesions / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D032448:Myoblasts, Skeletal / Q000166:cytology* / Q000187:drug effects; D014945:Wound Healing / Q000187:drug effects / Q000502:physiology; D054460:rho-Associated Kinases / Q000378:metabolism
D024042:Collagen Type I; D058575:Decorin; D005353:Fibronectins; C515854:Rock1 protein, mouse; C515855:Rock2 protein, mouse; D054460:rho-Associated Kinases