Evolution of tissue damage in compressive spinal cord injury in rats

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✓ The evolution of tissue damage in compressive spinal cord injuries in rats was studied using an immunohistochemical technique and by sodium dodecyl sulfate-polyacrylamide gel electropheresis (SDS-PAGE) analysis. The rupture of small vessels accompanied by intense tissue permeation of serum components in and around the hemorrhagic foci appeared to be immediate consequences of the mechanical insult. The loss of cell membrane integrity in neural elements became evident within 1 hour after injury as shown by the diffuse albumin-immunoreactivity of the cytoplasm. At the site of mechanical insult, approximately 30% of the neurofilament proteins were degraded within 1 hour, and 70% of them were lost within 4 hours after injury. A large number of cells positive for glial fibrillary acidic protein were found to demarcate the injured tissue within 1 hour after injury. The progression of tissue damage largely subsided within 48 hours. One week after, injury, severe degeneration of the ascending tracts in the posterior funiculus was shown clearly by axon staining and less convincingly by myelin staining. Secondary degeneration of the corticospinal tract in distal segments remained inconspicuous for up to 3 months.

Article Information

Address reprint requests to: Yuzo Iwasaki, M.D., Department of Neuropathology, Institute of Brain Diseases, Tohoku University School of Medicine, 1-1 Seiryo-Machi, Sendai 980, Japan.

© AANS, except where prohibited by US copyright law.

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Figures

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    A: Photomicrograph of a hemorrhagic necrotic lesion that developed less than 10 minutes after a 30-second compression with a vascular clip. H & E, × 25. B: A high-power view of the area indicated by them arrow in A. A fibrin deposit (arrow) is seen at the edge of a ruptured capillary which has been emptied after perfusion with a fixative solution. H & E, × 250. C: Photomicrograph showing extensive permeation of serum albumin in and around the hemorrhagic tissue, revealed by the avidin-biotin complex (ABC) staining method with anti-rat-albumin serum in a section adjacent to that in A. × 25. D: A high-power view of the area indicated by the arrow in C. The majority of vessels are emptied after transcardial perfusion. Note the lack of albumin-immunoreactivity in neurons (arrows). v = vein. ABC, × 125.

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    Photomicrograph showing strongly albumin-immunoreactive neuronal cells in the vicinity of a hemorrhagic lesion 1 hour after injury. Avidin-biotin complex method, × 55.

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    Photomicrographs of the pathology revealed by the avidin-biotin complex method with anti-neurofilament protein (NFP) sera. A: Specimen obtained 10 minutes after injury showing immediate loss of NFP-immunoreactivity from the tissue to which pressure was directly applied. Anti-200k stain, × 25. B: Specimen obtained 8 hours after compression. Secondary loss of NFP-immunoreactivity is apparently associated with tissue edema and it is less sharply demarcated than the lesion in A. Anti-200k stain, × 25. C: Swelling of axon terminals in the adjacent gray matter is seen in this specimen obtained 1 hour after injury. Strongly immunoreactive materials (arrows) are scattered within the neuropil. Anti-150k stain, × 125.

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    Photomicrographs showing astroglial reaction at the site of injury. Avidin-biotin complex with anti-GFAP (glial fibrillary acidic protein) serum, c = central canal; H = hemorrhagic lesion. A: The lesion is demarcated with GFAP-positive cells within 1 hour after injury. × 25. B: At 48 hours after injury, the surrounding tissue is paved with GFAP-positive cells. × 25.

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    Photomicrographs of the posterior funiculus in the distal segments 1 week after injury. A distinct wedge-shaped lesion is revealed by the Bodian stain (a) in the most dorsal portion of the cervical cord, but it is less conspicuous with Klüver-Barrera staining (b). Only loosening of nerve bundles and slight hypercellularity are suspected in the lumbar cord, as shown by a Bodian stain (c) and Klüver-Barrera stain (d). a and b: × 50; c and d: × 100.

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    Photomicrograph showing diffuse gliosis in the otherwise normal gray matter of the lumbar cord 1 week after injury. D = posterior funiculus. GFAP stain, × 30.

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    Sequential 7.5% SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electropheresis) profiles of the cytoskeletal proteins extracted from the injured tissue at various times after injury. Note the rapid degradation of neurofilament protein (NFP) with relative sparing of glial fibrillary acidic protein (GFAP).

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    Results of gel scanning of two time-course experiments on cytoskeletal protein degradation at the site of injury. Each experiment is indicated by closed and open symbols: stars = GFAP; circles = 68k; triangles = 150k; and squares = 200k.

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