title: Concentration-dependent requirement for local protein synthesis in motor neuron subtype-specific response to axon guidance cues.
abstract: Formation of functional motor circuits relies on the ability of distinct spinal motor neuron subtypes to project their axons with high precision to appropriate muscle targets. While guidance cues contributing to motor axon pathfinding have been identified, the intracellular pathways underlying subtype-specific responses to these cues remain poorly understood. In particular, it remains controversial whether responses to axon guidance cues depend on axonal protein synthesis. Using a growth cone collapse assay, we demonstrate that mouse embryonic stem cell-derived spinal motor neurons (ES-MNs) respond to ephrin-A5, Sema3f, and Sema3a in a concentration-dependent manner. At low doses, ES-MNs exhibit segmental or subtype-specific responses, while this selectivity is lost at higher concentrations. Response to high doses of semaphorins and to all doses of ephrin-A5 is protein synthesis independent. In contrast, using microfluidic devices and stripe assays, we show that growth cone collapse and guidance at low concentrations of semaphorins rely on local protein synthesis in the axonal compartment. Similar bimodal response to low and high concentrations of guidance cues is observed in human ES-MNs, pointing to a general mechanism by which neurons increase their repertoire of responses to the limited set of guidance cues involved in neural circuit formation.
authors.affiliation: Department of Pathology and Cell Biology, Center for Motor Neuron Biology and Disease, College of Physicians and Surgeons, PS 14-434, Columbia University, New York, New York 10032, USA.
authors.forename: Stéphane
authors.initials: S
authors.lastname: Nédelec
authors.forename: Mirza
authors.initials: M
authors.lastname: Peljto
authors.forename: Peng
authors.initials: P
authors.lastname: Shi
authors.forename: Mackenzie W
authors.initials: MW
authors.lastname: Amoroso
authors.forename: Lance C
authors.initials: LC
authors.lastname: Kam
authors.forename: Hynek
authors.initials: H
authors.lastname: Wichterle
mesh_terms: D000818:Animals; D001369:Axons / Q000378:metabolism / Q000502:physiology*; D002478:Cells, Cultured; D003463:Cues; D053595:Embryonic Stem Cells / Q000378:metabolism / Q000502:physiology; D036386:Ephrin-A5 / Q000008:administration & dosage / Q000502:physiology; D020439:Growth Cones / Q000473:pathology / Q000502:physiology; D006801:Humans; D008297:Male; D008565:Membrane Proteins / Q000008:administration & dosage / Q000502:physiology; D051379:Mice; D009046:Motor Neurons / Q000145:classification / Q000502:physiology*; D009419:Nerve Tissue Proteins / Q000008:administration & dosage / Q000502:physiology; D014176:Protein Biosynthesis / Q000502:physiology*; D040121:Semaphorin-3A; D015398:Signal Transduction / Q000502:physiology; D013116:Spinal Cord / Q000166:cytology / Q000502:physiology
chemical_list: D036386:Ephrin-A5; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; C491229:Sema3f protein, mouse; D040121:Semaphorin-3A