title: Proteomic analysis of C2C12 myoblast and myotube exosome-like vesicles: a new paradigm for myoblast-myotube cross talk?
abstract: Exosomes are nanometer-sized microvesicles formed in multivesicular bodies (MVBs) during endosome maturation. Exosomes are released from cells into the microenvironment following fusion of MVBs with the plasma membrane. During the last decade, skeletal muscle-secreted proteins have been identified with important roles in intercellular communications. To investigate whether muscle-derived exosomes participate in this molecular dialog, we determined and compared the protein contents of the exosome-like vesicles (ELVs) released from C2C12 murine myoblasts during proliferation (ELV-MB), and after differentiation into myotubes (ELV-MT). Using a proteomic approach combined with electron microscopy, western-blot and bioinformatic analyses, we compared the protein repertoires within ELV-MB and ELV-MT. We found that these vesicles displayed the classical properties of exosomes isolated from other cell types containing components of the ESCRT machinery of the MVBs, as well as numerous tetraspanins. Specific muscle proteins were also identified confirming that ELV composition also reflects their muscle origin. Furthermore quantitative analysis revealed stage-preferred expression of 31 and 78 proteins in ELV-MB and ELV-MT respectively. We found that myotube-secreted ELVs, but not ELV-MB, reduced myoblast proliferation and induced differentiation, through, respectively, the down-regulation of Cyclin D1 and the up-regulation of myogenin. We also present evidence that proteins from ELV-MT can be incorporated into myoblasts by using the GFP protein as cargo within ELV-MT. Taken together, our data provide a useful database of proteins from C2C12-released ELVs throughout myogenesis and reveals the importance of exosome-like vesicles in skeletal muscle biology.
authors.affiliation: CarMeN Laboratory (INSERM 1060, INRA 1362, INSA) University of Lyon, Faculté de Médecine Lyon-Sud, Oullins, France.
authors.forename: Alexis
authors.initials: A
authors.lastname: Forterre
authors.affiliation: CarMeN Laboratory (INSERM 1060, INRA 1362, INSA) University of Lyon, Faculté de Médecine Lyon-Sud, Oullins, France.
authors.forename: Audrey
authors.initials: A
authors.lastname: Jalabert
authors.affiliation: CarMeN Laboratory (INSERM 1060, INRA 1362, INSA) University of Lyon, Faculté de Médecine Lyon-Sud, Oullins, France.
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Berger
authors.affiliation: CEA, IRTSV, Laboratoire Biologie à Grande Echelle, Grenoble, France ; INSERM, U1038, Grenoble, France ; Grenoble Alpes University, IRTSV, Laboratoire Biologie à Grande Echelle, Grenoble, France.
authors.forename: Mathieu
authors.initials: M
authors.lastname: Baudet
authors.affiliation: CarMeN Laboratory (INSERM 1060, INRA 1362, INSA) University of Lyon, Faculté de Médecine Lyon-Sud, Oullins, France.
authors.forename: Karim
authors.initials: K
authors.lastname: Chikh
authors.affiliation: Centre Commun d'Imagerie de Laënnec (CeCIL), SFR Santé Lyon-Est, University of Lyon, Lyon, France.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Errazuriz
authors.affiliation: CarMeN Laboratory (INSERM 1060, INRA 1362, INSA) University of Lyon, Faculté de Médecine Lyon-Sud, Oullins, France.
authors.forename: Joffrey
authors.initials: J
authors.lastname: De Larichaudy
authors.affiliation: CarMeN Laboratory (INSERM 1060, INRA 1362, INSA) University of Lyon, Faculté de Médecine Lyon-Sud, Oullins, France.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Chanon
authors.affiliation: Centre de Génétique et de Physiologie Moléculaire et Cellulaire (CGPhiMC), CNRS UMR5534, University of Lyon, Lyon, France.
authors.forename: Michèle
authors.initials: M
authors.lastname: Weiss-Gayet
authors.affiliation: CEA, IRTSV, Laboratoire Biologie à Grande Echelle, Grenoble, France ; INSERM, U1038, Grenoble, France ; Grenoble Alpes University, IRTSV, Laboratoire Biologie à Grande Echelle, Grenoble, France.
authors.forename: Anne-Marie
authors.initials: AM
authors.lastname: Hesse
authors.affiliation: INSERM-UMR 1037, Centre de Recherche en Cancerologie de Toulouse (CRCT), Toulouse, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Record
authors.affiliation: CarMeN Laboratory (INSERM 1060, INRA 1362, INSA) University of Lyon, Faculté de Médecine Lyon-Sud, Oullins, France.
authors.forename: Alain
authors.initials: A
authors.lastname: Geloen
authors.affiliation: CarMeN Laboratory (INSERM 1060, INRA 1362, INSA) University of Lyon, Faculté de Médecine Lyon-Sud, Oullins, France.
authors.forename: Etienne
authors.initials: E
authors.lastname: Lefai
authors.affiliation: CarMeN Laboratory (INSERM 1060, INRA 1362, INSA) University of Lyon, Faculté de Médecine Lyon-Sud, Oullins, France.
authors.forename: Hubert
authors.initials: H
authors.lastname: Vidal
authors.affiliation: CEA, IRTSV, Laboratoire Biologie à Grande Echelle, Grenoble, France ; INSERM, U1038, Grenoble, France ; Grenoble Alpes University, IRTSV, Laboratoire Biologie à Grande Echelle, Grenoble, France.
authors.forename: Yohann
authors.initials: Y
authors.lastname: Couté
authors.affiliation: CarMeN Laboratory (INSERM 1060, INRA 1362, INSA) University of Lyon, Faculté de Médecine Lyon-Sud, Oullins, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Rome
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D055252:Cell-Derived Microparticles / Q000378:metabolism / Q000648:ultrastructure; D055354:Exosomes / Q000378:metabolism* / Q000648:ultrastructure; D051379:Mice; D018485:Muscle Fibers, Skeletal / Q000378:metabolism*; D032446:Myoblasts / Q000166:cytology / Q000378:metabolism*; D021381:Protein Transport; D020543:Proteome; D040901:Proteomics / Q000379:methods
chemical_list: D020543:Proteome