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

TY - JOUR

T1 - In Vitro Modeling of Nerve–Muscle Connectivity in a Compartmentalized Tissue Culture Device

AU - Barcellos Machado, Carolina

AU - Pluchon, Perrine

AU - Harley, Peter

AU - Rigby, Mark

AU - Gonzalez Sabater Budilov, Victoria

AU - Stevenson, Danielle C.

AU - Hynes, Stephanie

AU - Lowe, Andrew

AU - Burrone, Juan

AU - Viasnoff, Virgile

AU - Lieberam, Ivo

PY - 2019/7/31

Y1 - 2019/7/31

N2 - 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, three powerful tools have been merged—pluripotent stem cells, optogenetics, and microfabrication—and an open microdevice is designed in which motor axons grow from a neural compartment containing embryonic stem cell‐derived motor neurons and astrocytes through microchannels to form functional neuromuscular junctions with contractile myofibres in a separate compartment. Optogenetic entrainment of motor neurons in this reductionist neuromuscular circuit enhances neuromuscular junction formation more than twofold, mirroring the activity‐dependence of synapse development in vivo. An established motor neuron disease model is incorporated into the system and it is found that coculture of motor neurons with SOD1G93A astrocytes results in denervation of the central compartment and diminishes myofibre contractions, a phenotype which is rescued by the receptor interacting serine/threonine kinase 1 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.

AB - 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, three powerful tools have been merged—pluripotent stem cells, optogenetics, and microfabrication—and an open microdevice is designed in which motor axons grow from a neural compartment containing embryonic stem cell‐derived motor neurons and astrocytes through microchannels to form functional neuromuscular junctions with contractile myofibres in a separate compartment. Optogenetic entrainment of motor neurons in this reductionist neuromuscular circuit enhances neuromuscular junction formation more than twofold, mirroring the activity‐dependence of synapse development in vivo. An established motor neuron disease model is incorporated into the system and it is found that coculture of motor neurons with SOD1G93A astrocytes results in denervation of the central compartment and diminishes myofibre contractions, a phenotype which is rescued by the receptor interacting serine/threonine kinase 1 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.

KW - embryonic stem cell

KW - microdevice

KW - motor neuron

KW - myofibre

KW - optogenetics

UR - http://www.scopus.com/inward/record.url?scp=85065389691&partnerID=8YFLogxK

U2 - 10.1002/adbi.201800307

DO - 10.1002/adbi.201800307

M3 - Article

SN - 2366-7478

VL - 3

SP - 1

EP - 14

JO - Advanced Biosystems

JF - Advanced Biosystems

IS - 7

M1 - 1800307

ER -