title: PKCθ signaling is required for myoblast fusion by regulating the expression of caveolin-3 and β1D integrin upstream focal adhesion kinase.
abstract: Fusion of mononucleated myoblasts to form multinucleated myofibers is an essential phase of skeletal myogenesis, which occurs during muscle development as well as during postnatal life for muscle growth, turnover, and regeneration. Many cell adhesion proteins, including integrins, have been shown to be important for myoblast fusion in vertebrates, and recently focal adhesion kinase (FAK), has been proposed as a key mediator of myoblast fusion. Here we focused on the possible role of PKC, the PKC isoform predominantly expressed in skeletal muscle, in myoblast fusion. We found that the expression of PKC is strongly up-regulated following freeze injury-induced muscle regeneration, as well as during in vitro differentiation of satellite cells (SCs; the muscle stem cells). Using both PKC knockout and muscle-specific PKC dominant-negative mutant mouse models, we observed delayed body and muscle fiber growth during the first weeks of postnatal life, when compared with wild-type (WT) mice. We also found that myofiber formation, during muscle regeneration after freeze injury, was markedly impaired in PKC mutant mice, as compared with WT. This phenotype was associated with reduced expression of the myogenic differentiation program executor, myogenin, but not with that of the SC marker Pax7. Indeed in vitro differentiation of primary muscle-derived SCs from PKC mutants resulted in the formation of thinner myotubes with reduced numbers of myonuclei and reduced fusion rate, when compared with WT cells. These effects were associated to reduced expression of the profusion genes caveolin-3 and β1D integrin and to reduced activation/phosphorylation of their up-stream regulator FAK. Indeed the exogenous expression of a constitutively active mutant form of PKC in muscle cells induced FAK phosphorylation. Moreover pharmacologically mediated full inhibition of FAK activity led to similar fusion defects in both WT and PKC-null myoblasts. We thus propose that PKC signaling regulates myoblast fusion by regulating, at least in part, FAK activity, essential for profusion gene expression.
authors.affiliation: Department of Anatomy, Sapienza University of Rome, Rome, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Madaro
authors.forename: Valeria
authors.initials: V
authors.lastname: Marrocco
authors.forename: Piera
authors.initials: P
authors.lastname: Fiore
authors.forename: Paola
authors.initials: P
authors.lastname: Aulino
authors.forename: Piera
authors.initials: P
authors.lastname: Smeriglio
authors.forename: Sergio
authors.initials: S
authors.lastname: Adamo
authors.forename: Mario
authors.initials: M
authors.lastname: Molinaro
authors.forename: Marina
authors.initials: M
authors.lastname: Bouché
mesh_terms: D000818:Animals; D051244:Caveolin 3 / Q000235:genetics / Q000378:metabolism*; D002450:Cell Communication; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002459:Cell Fusion; D002478:Cells, Cultured; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D022001:Focal Adhesions / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D055786:Gene Knockout Techniques; D039641:Integrin beta Chains / Q000235:genetics / Q000378:metabolism*; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D024510:Muscle Development / Q000502:physiology; D032446:Myoblasts / Q000166:cytology / Q000378:metabolism*; D018008:Myogenin / Q000235:genetics / Q000378:metabolism; D051763:PAX7 Transcription Factor / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D051745:Protein Kinase C-delta / Q000172:deficiency / Q000235:genetics; D012038:Regeneration; D015398:Signal Transduction / Q000235:genetics*; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*
chemical_list: D051244:Caveolin 3; D039641:Integrin beta Chains; D018008:Myogenin; D051763:PAX7 Transcription Factor; C497322:Pax7 protein, mouse; D051416:Focal Adhesion Protein-Tyrosine Kinases; D051745:Protein Kinase C-delta