In Vitro Modeling of Nerve–Muscle Connectivity in a Compartmentalized Tissue Culture Device

Carolina Barcellos Machado, Perrine Pluchon, Peter Harley, Mark Rigby, Victoria Gonzalez Sabater, Danielle C. Stevenson, Stephanie Hynes, Andrew Lowe, Juan Burrone, Virgile Viasnoff, Ivo Lieberam
2019 Advanced Biosystems  
Motor neurons project axons from the hindbrain and spinal cord to muscle, where they induce myofibre contractions through neurotransmitter release at neuromuscular junctions. Studies of neuromuscular junction formation and homeostasis have been largely confined to in vivo models. In this study we have merged three powerful tools - pluripotent stem cells, optogenetics and microfabrication - and designed an open microdevice in which motor axons grow from a neural compartment containing embryonic
more » ... tem cell-derived motor neurons and astrocytes through microchannels to form functional neuromuscular junctions with contractile myofibers in a separate compartment. Optogenetic entrainment of motor neurons in this reductionist neuromuscular circuit enhanced neuromuscular junction formation more than two-fold, mirroring the activity-dependence of synapse development in vivo. We incorporated an established motor neuron disease model into our system and found that coculture of motor neurons with SOD1G93A astrocytes resulted in denervation of the central compartment and diminished myofiber contractions, a phenotype which was rescued by the Receptor Interacting Serine/Threonine Kinase 1 (RIPK1) inhibitor Necrostatin. This coculture system replicates key aspects of nerve-muscle connectivity in vivo and represents a rapid and scalable alternative to animal models of neuromuscular function and disease.
doi:10.1002/adbi.201800307 pmid:31428672 pmcid:PMC6699992 fatcat:xesgqsxnf5am5ddg2shvxrexe4