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J Neurophysiol 84: 1763-1769, 2000;
0022-3077/00 $5.00
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The Journal of Neurophysiology Vol. 84 No. 4 October 2000, pp. 1763-1769
Copyright ©2000 by the American Physiological Society

Control of Membrane Sealing in Injured Mammalian Spinal Cord Axons

Riyi Shi, Tomoko Asano, Neil C. Vining, and Andrew R. Blight

Division of Neurosurgery, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599

Shi, Riyi, Tomoko Asano, Neil C. Vining, and Andrew R. Blight. Control of Membrane Sealing in Injured Mammalian Spinal Cord Axons. J. Neurophysiol. 84: 1763-1769, 2000. The process of sealing of damaged axons was examined in isolated strips of white matter from guinea pig spinal cord by recording the "compound membrane potential," using a sucrose-gap technique, and by examining uptake of horseradish peroxidase (HRP). Following axonal transection, exponential recovery of membrane potential occurred with a time constant of 20 ± 5 min, at 37°C, and extracellular calcium activity ([Ca2+]o) of 2 mM. Most axons excluded HRP by 30 min following transection. The rate of sealing was reduced by lowering calcium and was effectively blocked at [Ca2+]o <=  0.5 mM, under which condition most axons continued to take up HRP for more than 1 h. Sealing at higher [Ca2+]o was blocked by calpain inhibitors (calpeptin and calpain inhibitor-1) indicating a requirement for type II (mM) calpain in the sealing process. Following compression injury, the amplitude of the maximal compound action potential conducted through the injury site was reduced. The extent of amplitude reduction was increased when the tract was superfused with calcium-free Krebs' solution (Ca2+ replaced by Mg2+). These results suggest that the fall in [Ca2+]o seen following injury in vivo is sufficient to prevent membrane sealing and may paradoxically contribute to axonal dieback, retrograde cell death, and "secondary" axonal disruption.




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