title: Axonal and neuromuscular synaptic phenotypes in Wld(S), SOD1(G93A) and ostes mutant mice identified by fiber-optic confocal microendoscopy.
abstract: We used live imaging by fiber-optic confocal microendoscopy (CME) of yellow fluorescent protein (YFP) expression in motor neurons to observe and monitor axonal and neuromuscular synaptic phenotypes in mutant mice. First, we visualized slow degeneration of axons and motor nerve terminals at neuromuscular junctions following sciatic nerve injury in Wld(S) mice with slow Wallerian degeneration. Protection of axotomized motor nerve terminals was much weaker in Wld(S) heterozygotes than in homozygotes. We then induced covert modifiers of axonal and synaptic degeneration in heterozygous Wld(S) mice, by N-ethyl-N-nitrosourea (ENU) mutagenesis, and used CME to identify candidate mutants that either enhanced or suppressed axonal or synaptic degeneration. From 219 of the F1 progeny of ENU-mutagenized BALB/c mice and thy1.2-YFP16/Wld(S) mice, CME revealed six phenodeviants with suppression of synaptic degeneration. Inheritance of synaptic protection was confirmed in three of these founders, with evidence of Mendelian inheritance of a dominant mutation in one of them (designated CEMOP_S5). We next applied CME repeatedly to living Wld(S) mice and to SOD1(G93A) mice, an animal model of motor neuron disease, and observed degeneration of identified neuromuscular synapses over a 1-4day period in both of these mutant lines. Finally, we used CME to observe slow axonal regeneration in the ENU-mutant ostes mouse strain. The data show that CME can be used to monitor covert axonal and neuromuscular synaptic pathology and, when combined with mutagenesis, to identify genetic modifiers of its progression in vivo.
authors.affiliation: Euan MacDonald Centre for MND Research, The University of Edinburgh, George Square, Edinburgh EH89JZ, UK.
authors.forename: Frances
authors.initials: F
authors.lastname: Wong
authors.forename: Li
authors.initials: L
authors.lastname: Fan
authors.forename: Sara
authors.initials: S
authors.lastname: Wells
authors.forename: Robert
authors.initials: R
authors.lastname: Hartley
authors.forename: Francesca E
authors.initials: FE
authors.lastname: Mackenzie
authors.forename: Oyinlola
authors.initials: O
authors.lastname: Oyebode
authors.forename: Rosalind
authors.initials: R
authors.lastname: Brown
authors.forename: Derek
authors.initials: D
authors.lastname: Thomson
authors.forename: Michael P
authors.initials: MP
authors.lastname: Coleman
authors.forename: Gonzalo
authors.initials: G
authors.lastname: Blanco
authors.forename: Richard R
authors.initials: RR
authors.lastname: Ribchester
mesh_terms: D000818:Animals; D001369:Axons / Q000473:pathology / Q000502:physiology / Q000648:ultrastructure*; D004195:Disease Models, Animal; D004724:Endoscopy / Q000379:methods*; D005260:Female; D005336:Fiber Optic Technology / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D008822:Mice, Transgenic; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods*; D016472:Motor Neuron Disease / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D009046:Motor Neurons / Q000166:cytology / Q000378:metabolism; D009154:Mutation; D009416:Nerve Regeneration / Q000502:physiology; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D009469:Neuromuscular Junction / Q000473:pathology / Q000502:physiology / Q000648:ultrastructure*; D010641:Phenotype; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism*; D013569:Synapses / Q000473:pathology / Q000502:physiology / Q000648:ultrastructure; D014855:Wallerian Degeneration / Q000378:metabolism / Q000473:pathology
chemical_list: D008164:Luminescent Proteins; D009419:Nerve Tissue Proteins; C442580:Wld protein, mouse; C109415:SOD1 G93A protein; D013482:Superoxide Dismutase