Motoneuron survival after chronic and sequential peripheral nerve injuries in the rat.
OBJECTIVE
Surgical repair of peripheral nerves following chronic nerve injury is associated with poor axonal regeneration and outcome. An underlying possibility is that chronic injuries may increase motoneuron cell death. The hypothesis that substantial motoneuron death follows chronic and sequential nerve injuries was tested in adult rats in this study.


METHODS
Thirty adult male Lewis rats underwent bilateral multistage surgeries. At initial surgery, Fast Blue (FB) tracer was injected at a nerve-crush injury site in the right control femoral motor nerve. The left femoral motor nerve was transected at the same level and either capped to prevent regeneration (Group 1), or repaired to allow axonal regeneration and reinnervation of the target quadriceps muscle (Group 2) (15 rats in each group). After 8 weeks in 6 rats/group, the left femoral nerve was cut and exposed to FB just proximal to prior nerve capping or repair and the rats were evaluated for FB-labeled motoneuron counts bilaterally in the spinal cord (this was considered survival after initial injury). In the remaining 9 animals/group, the left nerve was recut (sequential injury), exposed to FB, and repaired to a fresh distal saphenous nerve stump to permit axonal regeneration. Following another 6 weeks, Fluoro-Gold, a second retrograde tracer, was applied to the cut distal saphenous nerve. This allowed us to evaluate the number of motoneurons that survived (maintained FB labeling) and the number of motoneurons that survived but that also regenerated axons (double labeled with FB and Fluoro-Gold).


RESULTS
A mean number of 350 and 392 FB-labeled motoneurons were found after 8 weeks of nerve injury on the right and the left sides, respectively. This indicated no significant cell death due to initial nerve injury alone. A similar number (mean 390) of motoneurons were counted at final end point at 14 weeks, indicating no significant cell death after sequential and chronic nerve injury. However, only 50% (mean 180) of the surviving motoneurons were double labeled, indicating that only half of the population regenerated their axons.


CONCLUSIONS
The hypothesis that significant motoneuron cell death occurs after chronic and or sequential nerve injury was rejected. Despite cell survival, only 50% of motoneurons are capable of exhibiting a regenerative response, consistent with our previous findings of reduced regeneration after chronic axotomy.
Xu
Qing-Gui
QG
Division of Neurosurgery, Department of Clinical Neurosciences and Hotchkiss Brain Institute, University of Calgary, Calgary, Alberta, Canada.
Forden
Joanne
J
Walsh
Sarah K
SK
Gordon
Tessa
T
Midha
Rajiv
R
D000578:Amidines; D000818:Animals; D001369:Axons / Q000502:physiology; D019771:Axotomy; D002452:Cell Count; D016923:Cell Death / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D002908:Chronic Disease; D005267:Femoral Nerve / Q000166:cytology* / Q000502:physiology; D005456:Fluorescent Dyes; D008297:Male; D009046:Motor Neurons / Q000166:cytology* / Q000502:physiology; D009186:Myelin Sheath / Q000502:physiology; D009409:Nerve Crush; D009416:Nerve Regeneration / Q000502:physiology; D010523:Peripheral Nervous System Diseases / Q000473:pathology* / Q000503:physiopathology*; D052097:Quadriceps Muscle / Q000294:innervation; D051381:Rats; D011917:Rats, Inbred Lew; D013266:Stilbamidines
C049774:2-hydroxy-4,4'-diamidinostilbene, methanesulfonate salt; D000578:Amidines; D005456:Fluorescent Dyes; D013266:Stilbamidines; C031455:diamidino compound 253-50