Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism
BBA reportDifferent reactivities of monoclonal antibodies to ganglioside lactones
Abstract
Six murine monoclonal antibodies were found to react with ganglioside GD2 lactone as well as purified ganglioside GD2. However, the reactivities of these antibodies to various ganglioside lactones were found to differ from each other. Four antibodies only reacted with GD2 lactones, while the other two cross-reacted with lactones of other gangliosides such as GD1b and GT1b.
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Cited by (19)
The localization of gangliosides in neurons of the central nervous system: The use of anti-ganglioside antibodies
1996, Biochimica et Biophysica Acta - Reviews on BiomembranesDevelopmental changes of ganglioside expressions in postnatal rat cerebellar cortex
1995, Brain ResearchWe previously described the differential distribution of gangliosides in adult rat brain as detected by specific antibodies. We report here the distribution of gangliosides during the development of postnatal rat cerebellum by an immunofluorescence technique with mouse monoclonal antibodies (mAbs). Eleven mAbs that specifically recognize each ganglioside were used. Our study revealed that the expression of each ganglioside changed dramatically during the development. GD3 and O-Ac-GD3 were expressed intensely in the external granular layer at 1, 5, and 10 days, whereas GD2 was firstly detected in the internal granular layer at 5 days and GD1b was diffusely detected throughout all layers of the cerebellar cortex at early postnatal days. GD2 and GD1b were more intensely expressed in the granular layer at 20, 30, and 80 days, suggesting that premature granule cells express GD3 and its derivative, O-Ac-GD3, whereas mature granule cells express GD2 and GD1b intensely. On the other hand, GM1 was exclusively detected in the external granular layer and the molecular layer at 1 and 5 days. The staining sites spread gradually from these outer layers into the internal granular layer and the white matter after 10 days. The positive cells in the external granular layer and the molecular layer appeared to be Bergmann glial cells and their radially ascending cytoplasmic processes. The intensity of the staining in these specialized astroglial cells decreased gradually during postnatal days. In contrast, the expression of GQ1b was very faint at birth, but gradually increased during the development and was detected intensely in the internal granular layer, particularly in the cerebellar glomeruli in adulthood, suggesting that GQ1b expression may be associated with synapse-related structures. The developmental changes of the expression of other gangliosides were also recognized in the postnatal rat cerebellum. These results suggest that specific gangliosides may play an important role in regulating the early events responsible for the orderly formation of the cerebellar cortex.
Optimal conditions to radiolabel (<sup>3</sup>H or <sup>14</sup>C) aminosugar-containing glycosphingolipids by de-N-acetylation and re-N-acetylation
1995, BBA - General SubjectsThe optimal conditions were examined for selective re-N-acetylation with14C or3H acetic anhydride of de-N-acetylated aminosugar-containing glycosphingolipids. Re-N-acetylation, which is nearly quantitative within 10 minutes in methanol, occurs selectively up to a maximal 100% yield when using a molar ratio of 5 mol of acetic anhydride per mole of aminosugar present in the glycosphingolipid. Above this molar ratio, it was observed some O-acytylation of carbohydrates which could be removed by mild alkali treatment. The method allows the choice of14C- or3H-labeling of glycosphingolipids with a final specific radioactivity which depends solely on the one of acetic anhydride. The binding of specific antibodies to glycosphingolipids, which was abolished upon de-N-acetylation, was again detectable after re-N-acetylation with radioactive acetic anhydride, suggesting that the native structures were recovered. This procedure of radiolabeling offers safety, rapidity and broad applicability to alkali-stable aminosugar-containing glycosphingolipids.
The chemical constitution of gangliosides of the vertebrate nervous system
1995, Behavioural Brain ResearchGlycosphingolipids are uniquely distinguished amongst the glycoconjugates by the apparently systematic structuring of their ceramide-linked carbohydrate moieties. These often highly complex oligosaccharides provide a structural repertoire that may vary considerably according to cell types and animal species. However, as a possible reflection of their specific functional role in the central nervous system, the brain glycosphingolipids of all vertebrates follow the same principles of carbohydrate structuring with only minor variations: the anabolically early addition of sialic acid to lactosylceramide (Galβ4GlcβCer → NeuAcα3Galβ4GlcβCer) in central nervous tissue results in the preferential formation of ‘gangliosides’, i.e., sialic acid-containing glycosphingolipids. Higher gangliosides result from extensions of sialo-lactosylceramide by addition of nucleotide-activated monosaccharides. In consequence, gangliosides of the vertebrate central nervous system consist of ceramide-linked sialo-oligosaccharides of varying chain length with a ganglio-series core carbohydrate, i.e., GalNAcβ4Galβ3GalNacβ4Galβ4Glcβ < 0. Substitution by mono-, bis-, or tris-sialo-groups may variably be at the galactosides and N-acetylgalactosaminide residues in 3- and 6-positions of the ganglio-series oligosaccharides, respectively. Ganglioside, which is derived by sialylation of galactosylceramide, NeuAcα3GalβCer, is a characteristic constitutent of glial cells. In nerve tissue, ganglioside of the lacto-(Galβ(3GlcNAcβ3Galβ)n)4Glcβ<) and the neolacto-series (Galβ(4GlcNAcβ3Galβ)n4Glc<) are more characteristic of vertebrate peripheral nerves and neuroectoderm-derived tumours. Recent studies using monoclonal antibodies have revealed that various single ganglioside components are specifically distributed in nervous tissues. This finding adds a new dimension to the earlier notion that gangliosides are involved in membrane related phenomena including cell to cell interactions, as well as, the modulation of signalling mechanisms.
Antibody responses to ganglio-series gangliosides in different strains of inbred mice
1992, Molecular ImmunologyWe studied antibody responses after immunization with ganglio-series gangliosides against 10 strains of inbred mice, including Balb/c, C57BL/6, A/J, C3H/HeN, C3H/HeJ, CBA/N, AKR/N, NZB/N, DBA/2 and nu/nu Balb/c. Twelve gangliosides having NeuAc as their sialic acid moiety (GM4, GM3, GM2, GM1, GD3, O-Ac-GD3, GD2, GD1a, GD1b, GT1a, GT1b and GQ1b), four gangliosides having NeuGc (GM3, GM2, GM1 and GD3) and four asialo-gangliosides (GA4, GA3, GA2 and GA1) were injected intravenously adsorbed to Salmonella minnesota. The antibody titers of the mice sera were determined by an enzyme-linked immunosorbent assay and an immune adherence assay. Antibody responses were found to depend not only on the ganglioside used as an immunogen but also on the mouse strain. Gangliosides having a trisaccharide sequence (NeuAcα2 → 8NeuAcα2 → 3Gal-) such as GD3, GD2, GDlb, GT1a and GQ1b, in particular O-Ac-GD3, induced high-titer antibody responses, whereas those having a disaccharide sequence (NeuAcα2 → 3Gal-) such as GM4, GM3, GM2, GM1, GDla and GTl1 induced low-liter antibody responses. On the other hand, gangliosides with NeuGc developed minimum titers. In contrast, asialo-gangliosides induced much higher responses than the corresponding gangliosides. The differences in ceramide portions of these gangliosides did not appear to be involved in inducing antibody responses. Mice could be divided into three groups according to the magnitude of their antibody responses: Group 1, those that produce the highest antibody responses (C3H/HeN and A/J); Group 2, those that demonstrate moderate antibody titers (Balb/c, C57BL/6, DBA/2 and nu/nu Balb/c); and Group 3, those that make minimum responses (AKR/N, C3H/HeJ, CBA/N and NZB/N). The pattern of reactivity to the various gangliosides was similar in all the strains tested.
A cell-type specific ganglioside of glomerular podocytes in rat kidney: An O-acetylated GD3
1992, Kidney InternationalA cell-type specific ganglioside of glomerular podocytes in rat kidney: An O-acetylated GD3. We recently described a monoclonal antibody (clone 27A) that detected a membrane antigen specific for glomerular podocytes in adult rat kidney. After binding in vivo, the antibodies induced rapid changes in the foot processes. Here we show that in other rat tissues the antigen is detectable only in cells of adrenal medulla, in some cells of neural or neural crest origin, and in 1 to 5% of the cells of a rat pheochromocytoma cell line PC-12. Attempts to isolate the antigen revealed that it is an acidic, sialic acid containing lipid, as shown by thin layer chromatography and immuno-overlay techniques. Further characterization of the gangliosides extracted from rat glomeruli, bovine kidney, rat adrenal glands, or from PC-12 cells by ion exchange, thin layer, and gas liquid chromatography identified the antigenic lipid as a modified disialosyllactosylceramide (GD3). The results of mild alkaline treatment or periodate oxidation of the antigenic ganglioside, as well as chemical O-acetylation studies of standard gangliosides, showed that the modified ganglioside is O-acetylated, most probably at the 9-carbon of its terminal sialic acid residue. To our knowledge this is the first report of cell-type specific expression of gangliosides in the kidney.