title: A system for studying mechanisms of neuromuscular junction development and maintenance.
abstract: The neuromuscular junction (NMJ), a cellular synapse between a motor neuron and a skeletal muscle fiber, enables the translation of chemical cues into physical activity. The development of this special structure has been subject to numerous investigations, but its complexity renders in vivo studies particularly difficult to perform. In vitro modeling of the neuromuscular junction represents a powerful tool to delineate fully the fine tuning of events that lead to subcellular specialization at the pre-synaptic and post-synaptic sites. Here, we describe a novel heterologous co-culture in vitro method using rat spinal cord explants with dorsal root ganglia and murine primary myoblasts to study neuromuscular junctions. This system allows the formation and long-term survival of highly differentiated myofibers, motor neurons, supporting glial cells and functional neuromuscular junctions with post-synaptic specialization. Therefore, fundamental aspects of NMJ formation and maintenance can be studied using the described system, which can be adapted to model multiple NMJ-associated disorders.
authors.affiliation: Myology Research Center, UM76-INSERM U974-CNRS FRE 3617 Sorbonne Universités, UPMC Université Paris 06, Paris, France vilmont.valerie@gmail.com edgargomes@medicina.ulisboa.pt.
authors.forename: Valérie
authors.initials: V
authors.lastname: Vilmont
authors.affiliation: Myology Research Center, UM76-INSERM U974-CNRS FRE 3617 Sorbonne Universités, UPMC Université Paris 06, Paris, France.
authors.forename: Bruno
authors.identifier: 0000-0002-1888-3898
authors.initials: B
authors.lastname: Cadot
authors.affiliation: FRE CNRS 3693 (U.N.I.C), Unité de Neuroscience, Information et Complexité CNRS, Bât. 33, 1 Ave de la Terasse, Gif sur Yvette 91198, France.
authors.forename: Gilles
authors.initials: G
authors.lastname: Ouanounou
authors.affiliation: Myology Research Center, UM76-INSERM U974-CNRS FRE 3617 Sorbonne Universités, UPMC Université Paris 06, Paris, France Instituto de Medicina Molecular, Faculdade de Medicina da Universidade de Lisboa, Lisboa, Portugal vilmont.valerie@gmail.com edgargomes@medicina.ulisboa.pt.
authors.forename: Edgar R
authors.identifier: 0000-0002-6941-4872
authors.initials: ER
authors.lastname: Gomes
mesh_terms: D000818:Animals; D048430:Cell Shape; D018920:Coculture Techniques; D005260:Female; D042541:Intracellular Space / Q000378:metabolism; D008564:Membrane Potentials; D051379:Mice; D009119:Muscle Contraction; D018485:Muscle Fibers, Skeletal / Q000378:metabolism; D009469:Neuromuscular Junction / Q000254:growth & development* / Q000378:metabolism; D009474:Neurons / Q000166:cytology; D009482:Neurophysiology / Q000379:methods*; D017207:Rats, Sprague-Dawley; D013116:Spinal Cord / Q000378:metabolism; D013569:Synapses / Q000378:metabolism
keywords: Co-culture; Differentiation; Myofiber; NMJ