title: Electrical stimulation accelerates neuromuscular junction formation through ADAM19/neuregulin/ErbB signaling in vitro.
abstract: The mechanism by which electrical stimulation affects formation of neuromuscular junctions (NMJs) remains unknown. NG108-15, a neural cell line, is commonly used in in vitro co-culture models of myotubes to observe synapse formation; therefore, we employed this model to observe the effects of electrical stimulation on NMJ formation. Initially, L6 cells were differentiated and NG108-15 cells were then added to the same culture dish. After 2 and 3 days of co-culture, the cells were electrically stimulated at 50 V and 0.5 Hz for 0, 5, 30, and 60 min (C, ES5, ES30, and ES60 groups, respectively) and were analyzed after co-culture for 4 days. Immunofluorescence experiments showed significantly increased aggregation of acetylcholine receptors and inhibition of neural outgrowth in the ES30 and ES60 groups. Furthermore, ADAM19 and phospho-ErbB3 were found to be specifically localized in co-cultured NG108-15 cells. Immunoblotting demonstrated that synapsin 1, ADAM19 precursor and its activated form, phospho-ErbB3, and ERK1 protein levels had increased in an electrical stimulation period-dependent manner. Thus, we found that electrical stimulation accelerated NMJ formation, possibly through activation of ADAM19/neuregulin/ErbB signaling in NG108-15 cells.
authors.affiliation: Division of Bio-Environmental Adaptation Sciences, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Fukazawa
authors.forename: Masaya
authors.initials: M
authors.lastname: Matsumoto
authors.forename: Takeshi
authors.initials: T
authors.lastname: Imura
authors.forename: Elham
authors.initials: E
authors.lastname: Khalesi
authors.forename: Teruyuki
authors.initials: T
authors.lastname: Kajiume
authors.forename: Yumi
authors.initials: Y
authors.lastname: Kawahara
authors.forename: Keiji
authors.initials: K
authors.lastname: Tanimoto
authors.forename: Louis
authors.initials: L
authors.lastname: Yuge
mesh_terms: D051722:ADAM Proteins / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D004558:Electric Stimulation / Q000379:methods*; D051379:Mice; D020880:Neuregulins / Q000378:metabolism*; D009469:Neuromuscular Junction / Q000502:physiology* / Q000648:ultrastructure; D009474:Neurons / Q000502:physiology* / Q000648:ultrastructure*; D018739:Oncogene Proteins v-erbB / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology
chemical_list: D020880:Neuregulins; D018739:Oncogene Proteins v-erbB; D051722:ADAM Proteins; C096116:Adam19 protein, mouse