Quantitative chemical analysis of demyelination in cultured mouse spinal cord

Roth, German A.; Bornstein, Murray B.

doi:10.1016/0165-3806(84)90145-7

Cited by 9 publications

(9 citation statements)

References 26 publications

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“…For lipid quantitation, samples of the homogenates were treated with 20 volumes of chloroform-methanol (1 : 1 vol/vol) and then partitioned with water. Individual glycolipids and cholesterol in the lower phases were determined using a high-performance thin-layer chromatographic (HPTLC), densitometric method (Roth and Bornstein, 1984).…”

Section: Biochemical Analysismentioning

confidence: 99%

Experimental autoimmune encephalomyelitis: Antigen-induced inhibition of biochemical and immunohistological alterations

et al. 1996

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A comprehensive biochemical, immunological, and histological study was undertaken during suppression of experimental autoimmune encephalomyelitis (EAE) induced by antigen‐specific inhibition of the immune response. Pretreatment of Wistar rats by intraperitoneal administration of low doses of saline‐soluble bovine myelin or myelin basic protein (MBP) but not with ovalbumin suppresses the appearance of the clinical symptoms of EAE induced by sensitization with bovine myelin in complete Freund's adjuvant. Analysis of the central nervous system (CNS) of animals pretreated with MBP or whole myelin shows inhibition of the diminution of MBP and 2′,3′‐cyclic nucleotide 3′‐phosphohydrolase (CNPase) activity observed in the EAE animals or in rats pretreated with ovalbumin. With respect to the CNS lipid content, these suppressive treatments abolish the increase in esterified cholesterol and partially revert the diminution in the content of cerebrosides and total cholesterol characteristic of the acute stage of the disease. Concomitantly, meningeal and parenchymal infiltration with mononuclear cells and deposits of immunoglobulins in the infiltrated regions as well as in spinal cord motor neurons were reduced. Analysis of the humoral response to myelin antigens shows that all EAE as well as treated animals developed antibodies to MBP and other myelin proteins. However, a higher incidence and level of these antibodies was observed in nontreated EAE animals and MBP‐ and ovalbumin‐treated rats, while rats treated with total bovine myelin showed a highly reduced humoral response. The present results indicate that intraperitoneal treatment with soluble forms of myelin antigens, concomitant with the suppression of the clinical symptoms of the disease, markedly reduces the biochemical and histological alterations occurring in EAE animals and produces changes in the autoimmune humoral response. © 1996 Wiley‐Liss, Inc.

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Section: Biochemical Analysismentioning

confidence: 99%

Experimental autoimmune encephalomyelitis: Antigen-induced inhibition of biochemical and immunohistological alterations

et al. 1996

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“…Myelin sheaths developed fusiform swellings with progressive distortions and fragmentation. These effects increased with time of exposure but were less than those anticipated from previous experiments using heterologous cells and serum under similar conditions (Bornstein and Iwanami, 1971;Roth et al, 1983;Roth and Bornstein, 1984). Only about 50-60% of cultures exposed to samples from animals with EAE showed myelin alterations.…”

Section: Resultsmentioning

confidence: 56%

Correlation Between 2′,3′‐Cyclic Nucleotide 3′‐Phosphohydrolase Activity and Demyelination In Vitro Using a Syngeneic System

Roth

Bornstein

Röyttä

et al. 1985

Journal of Neurochemistry

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Cultures of myelinated SJL/J fetal mouse spinal cord were incubated with serum and lymphoid cells from syngeneic animals with experimental allergic encephalomyelitis (EAE) induced by syngeneic spinal cord homogenate (SSCH) in complete Freund's adjuvant or others injected with complete Freund's adjuvant alone. After 24 or 48 h of exposure, demyelination was determined by light microscopic examination and quantification of 2',3'-cyclic nucleotide 3'-phosphohydrolase activity. Cultures exposed to spleen or lymph node cells from SSCH-sensitized animals showed the greatest alterations in myelin and decreases in 2',3'-cyclic nucleotide 3'-phosphohydrolase activity whereas serum from these animals had less effect. Cells and serum from complete Freund's adjuvant-injected control animals also induced structural changes in myelin that were significantly less than changes induced by cells and serum from animals with EAE. These experiments show that lymphoid cells and serum obtained from SJL/J mice with acute EAE affected myelin biochemistry and morphology in syngeneic CNS cultures.

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“…For this reason various biochemical markers of demyelination have been studied. These include CNPase (Weissbarth et al, 1980;Roth et al, 1985), cerebrosides (Roth et al, 1983;Roth and Bornstein, 1984), and MBP (Sheffield and Kim, 1977;Shin et al, 1984;Nishimura et al, 1986). Due to the differing amounts of myelinated fibres in the cultures, control and experimental values show a relatively wide range of variability.…”

Section: Discussionmentioning

confidence: 99%

An In Vitro Micromethod for the Quantitative Assessment of Central Demyelination

Watson

Najbauer

Owen

et al. 1988

Journal of Neurochemistry

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We report the development of a simple and reliable method for the study of demyelination in vitro based on the measurement of 2':3'-cyclic nucleotide 3'-phosphodiesterase in isolated myelin. Using only small quantities of myelin (equivalent to 100 micrograms of myelin protein) the system was tested under conditions that are believed to approximate those found at the site of an inflammatory demyelinating lesion. Treatment with a combination of trypsin, phospholipase A2, and lysophosphatidylcholine was used to evaluate the method. This microsystem has the potential not only for testing the myelinotoxicity of soluble factors but also for investigating the involvement of inflammatory cells in the demyelinating process. Myelin degradation by elicited peritoneal macrophages could be demonstrated at relatively high densities of these cells. Nylon wool purified lymph node T cells from myelin basic protein-primed SJL/J mice, after selective expansion with antigen and interleukin 2, failed to induce any significant myelin breakdown unless a limited number of syngeneic activated macrophages were also present. T cells from mice that had been inoculated with keyhole limpet haemocyanin failed to show any effect. The advantages of this technique over other in vitro systems are that it enables the study of demyelination using syngeneic sources of myelin and defined cell populations.

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Quantitative chemical analysis of demyelination in cultured mouse spinal cord

Cited by 9 publications

References 26 publications

Experimental autoimmune encephalomyelitis: Antigen-induced inhibition of biochemical and immunohistological alterations

Experimental autoimmune encephalomyelitis: Antigen-induced inhibition of biochemical and immunohistological alterations

Correlation Between 2′,3′‐Cyclic Nucleotide 3′‐Phosphohydrolase Activity and Demyelination In Vitro Using a Syngeneic System

An In Vitro Micromethod for the Quantitative Assessment of Central Demyelination

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