title: Electric Pulse Stimulation of Myotubes as an In Vitro Exercise Model: Cell-Mediated and Non-Cell-Mediated Effects.
abstract: Regular exercise has emerged as one of the best therapeutic strategies to prevent and treat type-2-diabetes. Exercise-induced changes in the muscle secretome, consisting of myokines and metabolites, may underlie the inter-organ communication between muscle and other organs. To investigate this crosstalk, we developed an in vitro system in which mouse C2C12 myotubes underwent electric pulse stimulation (EPS) to induce contraction. Subsequently the effects of EPS-conditioned media (EPS-CM) on hepatocytes were investigated. Here, we demonstrate that EPS-CM induces Metallothionein 1/2 and Slc30a2 gene expression and reduces Cyp2a3 gene expression in rat hepatocytes. When testing EPS-CM that was generated in the absence of C2C12 myotubes (non-cell EPS-CM) no decrease in Cyp2a3 expression was detected. However, similar inductions in hepatic Mt1/2 and Slc30a2 expression were observed. Non-cell EPS-CM were also applied to C2C12 myotubes and compared to C2C12 myotubes that underwent EPS: here changes in AMPK phosphorylation and myokine secretion largely depended on EPS-induced contraction. Taken together, these findings indicate that EPS can alter C2C12 myotube function and thereby affect gene expression in cells subjected to EPS-CM (Cyp2a3). However, EPS can also generate non-cell-mediated changes in cell culture media, which can affect gene expression in cells subjected to EPS-CM too. While EPS clearly represents a valuable tool in exercise research, care should be taken in experimental design to control for non-cell-mediated effects.
authors.affiliation: Molecular Cancer Research and Center for Molecular Medicine, University Medical Centre Utrecht, Universiteitsweg 100, 3584 CG, Utrecht, The Netherlands.
authors.forename: Inkie J A
authors.initials: IJ
authors.lastname: Evers-van Gogh
authors.affiliation: Nutrition, Metabolism and Genomics Group, Division of Human Nutrition, Wageningen University, PO BOX 8129, 6700 EV Wageningen, the Netherlands.
authors.forename: Sheril
authors.initials: S
authors.lastname: Alex
authors.affiliation: 1] Nutrition, Metabolism and Genomics Group, Division of Human Nutrition, Wageningen University, PO BOX 8129, 6700 EV Wageningen, the Netherlands [2] Department of Medicine, Radboud University Medical Centre, Geert Grooteplein 8, P.O. Box 9101, Nijmegen 6500 HB, The Netherlands.
authors.forename: Rinke
authors.initials: R
authors.lastname: Stienstra
authors.affiliation: Molecular Cancer Research and Center for Molecular Medicine, University Medical Centre Utrecht, Universiteitsweg 100, 3584 CG, Utrecht, The Netherlands.
authors.forename: Arjan B
authors.initials: AB
authors.lastname: Brenkman
authors.affiliation: Nutrition, Metabolism and Genomics Group, Division of Human Nutrition, Wageningen University, PO BOX 8129, 6700 EV Wageningen, the Netherlands.
authors.forename: Sander
authors.initials: S
authors.lastname: Kersten
authors.affiliation: Molecular Cancer Research and Center for Molecular Medicine, University Medical Centre Utrecht, Universiteitsweg 100, 3584 CG, Utrecht, The Netherlands.
authors.forename: Eric
authors.initials: E
authors.lastname: Kalkhoven
mesh_terms: D000818:Animals; D002460:Cell Line; D004558:Electric Stimulation; D005786:Gene Expression Regulation; D051379:Mice; D009119:Muscle Contraction; D018485:Muscle Fibers, Skeletal / Q000166:cytology / Q000378:metabolism*; D009124:Muscle Proteins / Q000378:metabolism*; D051381:Rats
chemical_list: D009124:Muscle Proteins