Abstract
The present investigation introduces an in vitro model to study macrophage properties during demyelination. Rat dorsal root ganglia (DRG) were cultured for obtaining myelinated peripheral nerve fibers. These cultures were exposed to non-resident macrophages. In untreated control cultures, there was no indication of myelin removal by the added macrophages. DRG were exposed to enzymatically generated oxygen radicals using the xanthin/xanthin oxidase or the glucose/glucose oxidase system. Assessment of Schwann cell viability and ultrastructural morphology revealed different patterns of cell cytotoxicity and morphological changes in different experiments. High concentrations caused complete tissue necrosis of the DRG, while low concentrations did not affect either cell viability or ultrastructural morphology. Under intermediate experimental conditions, oxygen radicals caused non-lethal Schwann cell damage leading to Schwann cell retraction and myelin sheath rejection. Myelin lamellae were disrupted and decompacted. These changes were followed by a selective macrophage attack on myelin sheats, resulting in demyelination. Axons, Schwann cells and sensory ganglion cells survived this attack. The specificity of the oxygen radical effects was tested in experiments using the oxygen radical scavengers catalase and superoxide dismutase. Catalase prevented the described effects on cell morphology and subsequently blocked demyelination by non-resident macrophages.
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Armati PJ, Bonner J (1990) A technique for promoting Schwann cell growth from fresh and frozen biopsy nerve utilizing D-valine medium. In Vitro Cell Dev Biol 26: 1116–1118
Armati PJ, Constable AL, Llewellyn F (1990) A new medium for in vitro peripheral nervous tissue myelination without the use of antimitotics. J Neurosci Methods 33: 149–155
Beuche W, Friede RL (1984) The role of non-resident cells in Wallerian degeneration. J Neurocytol 13: 767–796
Brück W, Friede RL (1989) Activation of macrophages by recombinant interferon-gamma has no effect on myelin phagocytosis but hinders invasion of nerves in organ culture. J Neuroimmunol 25: 47–55
Brück W, Friede RL (1990) L-Fucosidase treatment blocks myelin phagocytosis by macrophages in vitro. J Neuroimmunol 27: 217–227
Brück W, Friede RL (1990) Anti-macrophage CR3 antibody blocks myelin phagocytosis by macrophages in vitro. Acta Neuropathol 80: 415–418
Brück W, Friede RL (1991) The role of complement in myelin phagocytosis during PNS Wallerian degeneration. J Neurol Sci 103: 182–187
Bunge MB, Bunge RP, Peterson ER, Murray MR (1967) A light and electron microscope study of long-term organized cultures of rat dorsal root ganglia. J Cell Biol 32: 439–466
Eldridge CF, Bunge MB, Bunge RP, Wood PM (1987) Differentiation of axon-related Schwann cells in vitro. I. Ascorbic acid regulates basal lamina assembly and myelin formation. J Cell Biol 105: 1023–1034
Griffin JW, Stoll G, Li CY, Tyor W, Cornblath DR (1990) Macrophage responses in inflammatory demyelinating neuropathies. Ann Neurol 27: S64-S68
Griffin JW, George R, Lobato C, Tyor WR, Yan LC, Glass JD (1992) Macrophage responses and myelin clearance during Wallerian degeneration: relevance to immune-mediated demyelination. J Neuroimmunol 40: 153–166
Hann PG, Beuche W, Neumann U, Friede RL (1988) The rate of Wallerian degeneration in the absence of immunoglobulins. A study in chick and mouse peripheral nerve. Brain Res 451: 126–132
Hann Bonnekoh PG, Scheidt P, Friede RL (1989) Myelin phagocytosis by peritoneal macrophages in organ cultures of mouse peripheral nerve. A new model for studying myelin phagocytosis in vitro. J Neuropathol Exp Neurol 48: 140–153
Hartung H-P, Toyka KV (1990) T-cell and macrophage activation in experimental autoimmune neuritis and Guillain-Barré syndrome. Ann Neurol 27 [Suppl]: S57-S63
Hartung H-P, Schäfer B, Heininger K, Stoll G, Toyka VK (1988) The role of macrophages and eicosanoids in the pathogenesis of experimental allergic neuritis. Brain 111: 1039–1059
Hartung H-P, Schäfer B, Heininger K, Toyka KV (1988) Suppression of experimental autoimmune neuritis by the oxygen radical scavengers superoxide dismutase and catalase. Ann Neurol 23: 453–460
Hartung H-P, Jung S, Stoll G, Zielasek J, Schmidt B, Archelos JJ, Toyka KV (1992) Inflammatory mediators in demyelinating disorders of the CNS and the PNS. J Neuroimmunol 40: 197–210
Heininger K, Schäfer B, Hartung H-P, Fierz W, Linington C, Toyka KV (1988) The role of macrophages in experimental autoimmune neuritis induced by a P2-specific T-cell line. Ann Neurol 23: 326–331
Jung S, Huitinga I, Schmidt B, Zielasek J, Dijkstra CD, Toyka KV, Hartung H-P (1993) Selective elimination of macrophages by dichlormethylene diphosphonate-containing liposomes suppresses experimental autoimmune neuritis. J Neurol Sci 119: 195–202
Konat GW, Wiggins RC (1985) Effect of reactive oxygen species on myelin membrane proteins. J Neurochem 45: 1113–1118
Lampert PW (1969) Mechanism of demyelination in experimental allergic neuritis. Lab Invest 20: 127–138
Lieberman AR (1976) Sensory ganglia. In: Landon DN (ed) The peripheral nerve. Chapman and Hall, London, pp 188–278
McCord JM, Fridovich I (1968) The reduction of cytochrome c by milk xanthine oxidase. J Biol Chem 243: 5753–5600
Mithen FA, Ilyas AA, Birchem R, Cook SD (1992) Effects of Guillain-Barré sera containing antibodies against glycolipids in cultures of rat Schwann cells and sensory axons. J Neurol Sci 112: 223–232
Rosen JL, Brown MJ, Hickey WF, Rostami A (1990) Early myelin lesions in experimental allergic neuritis. Muscle Nerve 13: 629–636
Scaravilli F, Giometto B, Chimelli L, Sinclair E (1991) Macrophages in human sensory ganglia: an immunohistochemical and ultrastructural study. J Neurocytol 20: 609–624
Scolding NJ, Houston AJ, Morgan BP, Campbell AK, Compston DAS (1989) Reversible injury of cultured rat oligodendrocytes by complement. Immunology 67: 441–445
Smith KJ, Hall SM, Schauf CL (1985) Vesicular demyelination induced by raised intracellular calcium. J Neurol Sci 71: 19–37
Stocker R, Peterhans E (1989) Antioxidant properties of conjugated bilirubin and biliverdin: biological relevant scavenging of hypochlorous acid. Free Radic Res Commun 6: 57–66
Stoll G, Hartung H-P (1992) The role of macrophages in degeneration and immune-mediated demyelination of the peripheral nervous system. Adv Neuroimmunol 2: 163–179
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Brück, W., Brück, Y., Diederich, U. et al. Dorsal root ganglia cocultured with macrophages: an in vitro model to study experimental demyelination. Acta Neuropathol 88, 459–464 (1994). https://doi.org/10.1007/BF00389499
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DOI: https://doi.org/10.1007/BF00389499