Rapid loss of motor nerve terminals following hypoxia-reperfusion injury occurs via mechanisms distinct from classic Wallerian degeneration.
Motor nerve terminals are known to be vulnerable to a wide range of pathological stimuli. To further characterize this vulnerability, we have developed a novel model system to examine the response of mouse motor nerve terminals in ex vivo nerve/muscle preparations to 2 h hypoxia followed by 2 h reperfusion. This insult induced a rapid loss of neurofilament and synaptic vesicle protein immunoreactivity at pre-synaptic motor nerve terminals but did not appear to affect post-synaptic endplates or muscle fibres. The severity of nerve terminal loss was dependent on the age of the mouse and muscle type: in 8-12-week-old mice the predominantly fast-twitch lumbrical muscles showed an 82.5% loss, whereas the predominantly slow-twitch muscles transversus abdominis and triangularis sterni showed a 57.8% and 27.2% loss, respectively. This was contrasted with a > 97% loss in the predominantly slow-twitch muscles from 5-6-week-old mice. We have also demonstrated that nerve terminal loss occurs by a mechanism distinct from Wallerian degeneration, as the slow Wallerian degeneration (Wld(s)) gene did not modify the extent of nerve terminal pathology. Together, these data show that our new model of hypoxia-reperfusion injury is robust and repeatable, that it induces rapid, quantitative changes in motor nerve terminals and that it can be used to further examine the mechanisms regulating nerve terminal vulnerability in response to hypoxia-reperfusion injury.
Centre for Integrative Physiology, School of Biomedical Sciences, University of Edinburgh, Old Medical School, Edinburgh, UK.
Becki
B
Baxter
Thomas H
TH
Gillingwater
Simon H
SH
Parson
D000818:Animals; D005260:Female; D007150:Immunohistochemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence; D023421:Models, Animal; D009046:Motor Neurons / Q000473:pathology*; D018656:Muscle Fibers, Fast-Twitch / Q000473:pathology; D018657:Muscle Fibers, Slow-Twitch / Q000473:pathology; D018482:Muscle, Skeletal / Q000473:pathology; D009410:Nerve Degeneration / Q000473:pathology*; D009469:Neuromuscular Junction / Q000473:pathology*; D017729:Presynaptic Terminals / Q000473:pathology*; D015427:Reperfusion Injury / Q000473:pathology*; D013995:Time; D014855:Wallerian Degeneration