title: Experimental spinal cord injury: spatiotemporal characterization of elemental concentrations and water contents in axons and neuroglia.
abstract: To examine the role of axonal ion deregulation in acute spinal cord injury (SCI), white matter strips from guinea pig spinal cord were incubated in vitro and were subjected to graded focal compression injury. At several postinjury times, spinal segments were removed from incubation and rapidly frozen. X-ray microanalysis was used to measure percent water and dry weight elemental concentrations (mmol/kg) of Na, P, Cl, K, Ca, and Mg in selected morphological compartments of myelinated axons and neuroglia from spinal cord cryosections. As an index of axon function, compound action potentials (CAP) were measured before compression and at several times thereafter. Axons and mitochondria in epicenter of severely compressed spinal segments exhibited early (5 min) increases in mean Na and decreases in K and Mg concentrations. These elemental changes were correlated to a significant reduction in CAP amplitude. At later postcompression times (15 and 60 min), elemental changes progressed and were accompanied by alterations in compartmental water content and increases in mean Ca. Swollen axons were evident at all postinjury times and were characterized by marked element and water deregulation. Neuroglia and myelin in severely injured epicenter also exhibited significant disruptions. In shoulder areas (adjacent to epicenter) of severely injured spinal strips, axons and mitochondria exhibited modest increases in mean Na in conjunction with decreases in K, Mg, and water content. Following moderate compression injury to spinal strips, epicenter axons exhibited early (10 min postinjury) element and water deregulation that eventually recovered to near control values (60 min postinjury). Na(+) channel blockade by tetrodotoxin (TTX, 1 microM) perfusion initiated 5 min after severe crush diminished both K loss and the accumulation of Na, Cl, and Ca in epicenter axons and neuroglia, whereas in shoulder regions TTX perfusion completely prevented subcellular elemental deregulation. TTX perfusion also reduced Na entry in swollen axons but did not affect K loss or Ca gain. Thus graded compression injury of spinal cord produced subcellular elemental deregulation in axons and neuroglia that correlated with the onset of impaired electrophysiological function and neuropathological alterations. This suggests that the mechanism of acute SCI-induced structural and functional deficits are mediated by disruption of subcellular ion distribution. The ability of TTX to reduce elemental deregulation in compression-injured axons and neuroglia implicates a significant pathophysiological role for Na(+) influx in SCI and suggests Na(+) channel blockade as a pharmacotherapeutic strategy.
authors.affiliation: Department of Anesthesiology, Albert Einstein College of Medicine, Montefiore Medical Center, Bronx, New York 10467, USA.
authors.forename: R M
authors.initials: RM
authors.lastname: LoPachin
authors.forename: C L
authors.initials: CL
authors.lastname: Gaughan
authors.forename: E J
authors.initials: EJ
authors.lastname: Lehning
authors.forename: Y
authors.initials: Y
authors.lastname: Kaneko
authors.forename: T M
authors.initials: TM
authors.lastname: Kelly
authors.forename: A
authors.initials: A
authors.lastname: Blight
mesh_terms: D000818:Animals; D001369:Axons / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D001834:Body Water / Q000378:metabolism*; D004573:Electrolytes / Q000378:metabolism*; D006168:Guinea Pigs; D066298:In Vitro Techniques; D009457:Neuroglia / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D013116:Spinal Cord / Q000378:metabolism* / Q000503:physiopathology; D013117:Spinal Cord Compression / Q000378:metabolism / Q000503:physiopathology; D013119:Spinal Cord Injuries / Q000378:metabolism* / Q000503:physiopathology*; D013779:Tetrodotoxin / Q000494:pharmacology; D013997:Time Factors; D014131:Trace Elements / Q000378:metabolism*
chemical_list: D004573:Electrolytes; D014131:Trace Elements; D013779:Tetrodotoxin