title: Neuromuscular contacts induce nitric oxide signals in skeletal myotubes in vitro.
abstract: It has previously been shown that skeletal myotubes express nitric oxide synthase (NOS) and produce and release NO signals. NOS is also part of agrin-induced acetylcholine receptor aggregations on myotubes. As nerve-muscle interactions underlie reciprocal signaling mechanisms, we hypothesized that NO signals in target myotubes may be induced by neuromuscular contacts in development. Chimeric neuron-myotube co-cultures were prepared using p75-selected spinal cord neurons from embryonic chicken. Confocal imaging revealed robust 1,2-diaminoanthraquinone red fluorescence indicative of de novo formation of NO only in those myotubes which were contacted by neurites, also verified by pre- and postsynaptic marker costaining (anti-synaptotagmin and alpha-bungarotoxin). Neither soluble agrin nor sensory dorsal root ganglionic neurons showed comparable effects in this model. We concluded that in target skeletal muscle cells the NOS/NO system is controlled by motoneuron contacts by as yet incompletely understood signaling mechanisms. Endogenous NO signaling in myotubes may be essential during synapse formation and plasticity of the neuromuscular system.
authors.affiliation: Neurobiology and Neuromuscular Working Group, Institute of Anatomy, Campus Benjamin Franklin, Charité-University Medicine, Berlin, Germany.
authors.forename: Britta
authors.initials: B
authors.lastname: Püttmann
authors.forename: Eva-Maria
authors.initials: EM
authors.lastname: Gerlach
authors.forename: Mark
authors.initials: M
authors.lastname: Krüger
authors.forename: Dieter
authors.initials: D
authors.lastname: Blottner
mesh_terms: D000818:Animals; D002038:Bungarotoxins / Q000378:metabolism; D002452:Cell Count / Q000379:methods; D002478:Cells, Cultured; D002642:Chick Embryo; D018920:Coculture Techniques / Q000379:methods; D004347:Drug Interactions; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005455:Fluorescent Antibody Technique / Q000379:methods; D005727:Ganglia, Spinal / Q000166:cytology; D007211:Indoles; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods; D032446:Myoblasts / Q000201:enzymology*; D009474:Neurons / Q000502:physiology*; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D011940:Receptor Aggregation / Q000187:drug effects / Q000502:physiology; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D013997:Time Factors
chemical_list: D002038:Bungarotoxins; D004791:Enzyme Inhibitors; D007211:Indoles; C007293:DAPI; D019001:Nitric Oxide Synthase