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Sulfatide synthesis: inhibition by experimental allergic encephalomyelitis serum.

The rate of (35)S incorporation into cerebroside sulfate in cultures of embryo mouse spinal cord shows a rapid acceleration at the time of myelin formation. Exposure of cultures to dilute serum from rabbits with experimental allergic encephalomyelitis results in almost complete inhibition of sulfatide synthesis. Within 24 hours after replacement of inhibiting medium with normal medium there is an increase in sulfatide synthesis followed by myelination.

Animals↗

Axons: isolation from mammalian central nervous system.

Centrifugation of a homogenate of white matter, in a solution of buffered sucrose containing salt, produces a floating layer of myelinated axons. When these are suspended in hypotonic buffer, the mnyelin swells and strips away from the axon. Axons are then separated from the myelin by centrifugation. The resulting preparation consists of a variable population of processes with lengths up to 200 micrometers and diameters between 0.3 and 5.0 micrometers. The axons contain neurofilaments and mitochondria, although no axolemma or neurotubules are evident. The preparation contains cerebroside and sulfatide, yet is essentially free of myelin.

Animals↗

Cholera toxin: interaction of subunits with ganglioside GM1.

Vibrio cholerae exotoxin is an aggregate of two different noninterconvertible subunits (molecular weights about 15,000 and about 25,000). Only the smaller subunit reacts with ganglioside G(M1), a possible biological receptor. The larger subunit, found only in active toxin molecules, can be eluted with 8 molar urea from insoluble complexes of toxin, ganglioside, and cerebroside.

Binding Sites↗

Myelination inhibiting and neuroelectric blocking factors in experimental allergic encephalomyelitis.

Sensitization of Lewis rats with whole central nervous system tissue or with purified myelin induced both experimental allergic encephalomyelitis (EAE) and a serum factor which inhibited myelin formation in vitro. Sensitization with the encephalitogenic factor, myelin basic protein, induced EAE, but not the myelination inhibition factor. Sensitization with cerebroside induced neither EAE nor myelination inhibition factor. The serums from control animals without EAE as well as from animals sensitized with all of the above antigens blocked evoked electrical responses in vitro.

Animals↗

Biochemical studies in Nevin-Jones syndrome.

The lipid profiles of the brain in four cases of Nevin-Jones syndrome are described. A loss of phosphatidyl ethanolamine was found in both white matter and cortex in two cases as well as a more general loss of lipids in the cortex in one patient; there was a normal cerebrosidesulphatide ratio in myelin. These findings indicate a general loss of myelin over a period of time. The distribution of fatty acids in cerebrosides and sulphatides show a slight reduction in nervonic acid (24:1). The findings support the view that a secondary demyelination is present.

Brain Diseases↗

Expression of glycolipids and myelin-associated glycoprotein during the differentiation of oligodendrocytes: comparison of the CG-4 glial cell line to primary cultures.

The expression of cerebrosides, sulfatides, gangliosides and the myelin-associated glycoprotein (MAG) during differentiation of the CG-4 line of oligodendrocyte progenitors [Louis et al.: J. Neurosci Res 31: 193, 1992] was compared with their expression in primary cultures of oligodendrocyte precursors [McCarthy and de Vellis: J Cell Biol 85: 890, 1980]. When the CG-4 cells differentiated from bipolar progenitors to oligodendrocytes, there was a decrease of glucosylcerebroside synthesis and an increase in galactosylcerebroside and sulfatide synthesis. However, even after differentiation, the incorporation of [3H]galactose into these glycolipids, the amounts of galactosylcerebroside and sulfatide, and the galactocerebroside/sulfatide ratio were all much less than in primary cultures of differentiating oligodendrocytes. The major gangliosides in differentiated primary oligodendrocyte cultures were GM3 and GD3, and GD3 was also a major ganglioside in the CG-4 line. However, unlike primary cultures of O-2A lineage cells in which GM3 synthesis increased dramatically during differentiation to oligodendrocytes, the CG-4 cells expressed very little GM3. Also, the CG-4 cells expressed larger amounts of more complex gangliosides, e.g. GD1b and GT1b, which were almost entirely restricted to the b-series. The amount of MAG expressed by the CG-4 cells increased substantially when they differentiated to oligodendrocytes, and it was almost all the large immature isoform. However, even after differentiation, the amount expressed was less than in differentiated primary oligodendrocyte cultures. Overall, the lower expression of myelin-related glycolipids and MAG by the CG-4 line suggests a lesser degree of differentiation in comparison to primary oligodendrocytes under the culture conditions of these experiments, but the larger amounts of cells available from the CG-4 line should be useful for investigating glycolipid and MAG function related to the early stages of myelinogenesis.

Animals↗

Incorporation of radioactivity from U-14C-glucose into oligodendrocytes and myelin of quaking mice and their littermate controls.

Oligodendrocytes and myelin were purified from the cerebra of quaking mice and their littermate controls (11-60 days of age) after injecting the animals intraperitoneally with U-14C-glucose. A peak of incorporation of radioactivity in the lipid extract of oligodendrocytes of both quaking and normal mice at 16-18 days of age was found, suggesting that the onset of myelination in the cerebra starts approximately at the same time for quaking mice and their littermate controls. Nevertheless the level of incorporation per cell was lower in the oligodendrocytes of quaking mice (50% of the control). The pattern of incorporation into myelin during development was similar between the two strains, but the specific activity as measured in dpm/mg protein was higher in the myelin of young quaking animals (up to 16 days). Peaks of incorporation were found in cerebrosides and sulfatides of oligodendrocytes and myelin in normal controls at 18 days. In the quaking mice these peaks were absent in oligodendrocytes and much delayed in the myelin of the mutant. The results would suggest that the defect in the quaking mutant in respect to myelination is in oligodendrocyte metabolism and thus in an early stage of the assembly of the myelin membrane.

Animals↗

Pelizaeus-Merzbacher disease: biochemical analysis of isolated myelin (electron-microscopy; protein, lipid and unsubstituted fatty acids analysis).

Analysis of myelin from a leukodystrophic brain was performed (Pelizaeus-Merzbacher disease, classical type). Myelin recovery was 7% of normal, when isolated by ultracentrifugation. Electron microscopy showed a great amount of loose lamellae, with less thick sheaths and periodicity close to normal. This myelin contains fewer lipids than normal, sphingolipids and plasmalogens being reduced. FAtty acids from phospholipids are essentially normal, however enols from plasmalogens are largely reduced. Purified sphingolipids (cerebrosides, sulfatides and sphingomyelin) present a considerable diminution in very long chain fatty acids; the ratio of very long chain fatty acids (over C18) on shorter chains is 1% of the normal value for saturated fatty acids and 2% for the monounsaturated homologues. Protein analysis showed that basic protein and proteolipids were reduced, Wolfgram proteins being relatively increased.

Brain Chemistry↗

Effect of undernutrition on lipid metabolism of brain. In vivo incorporation of labelled acetate and palmitate into lipids.

Albino rats were malnourished from neonatal to weanling stage by increasing the litter size. The in vivo incorporation of (1-(14)C)-acetate, (1-(14)C)-palmitate into the lipid classes was examined in the animals. A stimulated incorporation of the label was observed in total lipids, phospholipids, cholesterol, cerebrosides and sulfatides of the brain of malnourished animals compared to controls.

Acetates↗

Formation of 7-dehydrocholesterol-containing membrane rafts in vitro and in vivo, with relevance to the Smith-Lemli-Opitz syndrome.

Smith-Lemli-Opitz syndrome (SLOS) is a recessive disease typified by 7-dehydrocholesterol (7DHC) accumulation and depletion of cholesterol. Because cholesterol is a primary component of detergent-resistant membrane domains ("rafts"), we examined the compatibility of 7DHC with raft formation. Liposomes containing bovine brain phosphatidylcholine, sphingomyelin, cerebrosides, and either cholesterol, 7DHC, or coprostanol (the latter being incompatible with raft formation) were prepared. 7DHC was indistinguishable from cholesterol in its ability to become incorporated into membrane rafts, as judged by physical and chemical criteria, whereas coprostanol did not form rafts. The in vivo compatibility of 7DHC with raft formation was evaluated in brains of rats treated with trans-1,4-bis(2-dichlorobenzylamino-ethyl)cyclohexane dihydrochloride (AY9944), which mimics the SLOS biochemical defect. 7DHC/cholesterol ratios in rafts and whole brains from AY9944-treated rats were similar, indicating comparable efficiency of 7DHC and cholesterol incorporation into brain rafts. In contrast, dolichol (a nonsterol isoprenoid incompatible with raft formation) was greatly depleted in brain rafts relative to whole brain. Although brain raft fractions prepared from AY9944-treated and control rats yielded similar sterol-protein ratios, their gel electrophoresis profiles exhibited multiple differences, suggesting that altered raft sterol composition perturbs raft protein content. These results are discussed in the context of the SLOS phenotype, particularly with regard to the associated central nervous system defects.

Animals↗

Metachromatic leukodystrophy without arylsulfatase A deficiency.

Two siblings of consanguinous parents were noted to have a neurologic syndrome marked by developmental delay, regression of psychomotor performance, marked spasticity and progressive central nervous system degeneration. Markedly delayed nerve conduction times and a sural nerve biopsy which demonstrated changes typical of metachromatic leukodystrophy (MLD) were evident. Impairment of sulfated glycolipid metabolism was documented by analysis of glycospingolipid in urinary sediment. In spite of these findings, activities of arylsulfatase A and cerebroside sulfatidase in white blood cells and cultured skin fibroblasts were near normal. However, when intact growing fibroblasts were loaded with 35SO4-sulfatide a clear defect in sulfatide cleavage, comparable to that seen in MLD patients, was observed. Thus, these patients represent a new form of sulfatide storage disease -- MLD characterized by intact enzyme activity in cell homogenates but defective sulfolipid metabolism in vivo and in intact fibroblasts.

Adult↗

Fetal brain development in diabetic guinea pigs.

Fetal brain development was investigated near term in guinea pigs rendered diabetic with streptozotocin. The liver and the placenta were used as reference organs. Compared to controls, those fetuses from diabetic animals had normal cerebrum and cerebellum weights, but higher liver and placenta weights in relation to fetal weights. Although liver and placenta cell number (DNA content) was unchanged, it was significantly increased in the fetal cerebrum and cerebellum of diabetics. Although the tissue protein concentration was decreased in the liver and the placenta, it was unchanged and even increased in the cerebrum and cerebellum, respectively. The concentration of myelin (cerebroside-sulfatide) was unchanged in the cerebrum, but it was increased in the cerebellum of diabetic animals. These data suggest that diabetes has a growth-promoting effect on the fetal brain cell number. Furthermore, differences in the protein content between fetal organs may reflect abnormalities in protein metabolism which do not affect the brain during diabetic pregnancies.

Animals↗

Fruiting-inducing activity and antifungal properties of lipid components in members of Annelida.

Fruiting-inducing activity and antifungal properties of lipid components in the phylum Annelida were examined. Some amphoteric cerebrosides carrying a phosphocholine group showed fruiting-inducing activity on Schizophyllum commune, and one of them possessed activity comparable to that of Sch II, the most potent substance known. Furthermore, alkyl lysophosphatidylcholines were found to have an inhibitory effect on the growth of phytopathogenic fungi, Alternaria kikuchiana and Phomopsis mali. The relationship between structure and biological activities is discussed.

Alternaria↗

Influence of high-protein diets on brain phospholipid, ganglioside and monogalactosyl glycolipid content previously altered by long-term low-protein and ethanol administration.

A previous study was undertaken to determine the influence of the combined effects of ethanol consumption and a low-protein diet on brain membrane (phospholipids and gangliosides) and myelin (monogalactosyl glycolipids) lipids. Male Wistar rats chronically consuming ethanol were fed isoenergetic diets containing 22% protein (controls) or 6% protein (protein-deprived) from 60 to 240 days of age. Protein malnutrition initiated at 60 days of age affected phosphatidylglycerol, sphingophospholipid, monosialoganglioside GM1 and cerebroside containing nervonic fatty acid by 240 days of age. Among the effects of ethanol was an increase of the relative proportion of lipid galactose in monogalactosyl glycolipids and of N-acetylneuraminic acid in the trisialoganglioside GT1b, while there was decrease of the lipid phosphorus in the investigated phospholipids and of the disialoganglioside GD1a. The low-protein diet generally interfered with ethanol effects except for monosialo GM3, GM2 and GM1 gangliosides. The present study was carried out to investigate whether the ethanol-induced effect on these brain lipids could be elevated when we switched these animals to diets with 21 or 34% protein during the following 90 days. It was found that increasing protein intake and cessation of drinking ethanol returned the investigated brain phospholipids, monogalactosyl glycolipids and gangliosides to control values.

Animals↗

Effect of 3-acetylpyridine on the content of myelin in the rat brain.

Ten-week old rats were fed an AIN-76 purified die ad libitum and were given 3-acetylpyridine which is an antagonistic agent of nicotinic acid. The quantity and composition of myelin in the brain were analyzed after administration of 3-acetylpyridine for 40 days. Body weight, myelin yield, cerebroside level and specific activity of 2',3'-cyclic nucleotide-3'-phosphohydrolase in the brain decreased by administration of 3-acetylpyridine. Despite the decreased myelin yield, the proportion of protein and total lipid and the percentage composition of lipid in myelin did not change by administration of 3-acetylpyridine. Therefore, we have concluded that 3-acetylpyridine plays a significant role in the loss of myelin.

2',3'-Cyclic-Nucleotide Phosphodiesterases↗

Lipid composition in postnatal methylazoxymethanol-treated swiss albino mouse cerebellum.

Postnatal Swiss albino mice were treated with with methylazoxymethanol acetate (MAM) or saline and sacrificed at 25 days of age. Granule cell depletion resulted. Significant reduction in the cerebellar weight, protein, ganglioside sialic acid , cerebrosides, sulfatides, and phospholipids were documented. There was not, however, selective reduction of any component known to be associated with the synaptic structures. Specific association of cerebellar ganglioside with its granule cell population was not substantiated. Cerebroside/sulfatide ratios were not different in the two groups, indicating that significant alterations previously observed in spinal cord were not present in the cerebellum. It was concluded that the bulk of cerebellar granule cells and their synaptic connections could be deleted without affecting total ganglioside and phospholipid concentrations.

Animals↗

[Method of gradient chromatography for the isolation of purified lipid fractions from nerve tissue in studies using radioactive carbon, C14].

A method is developed for separation of lipids from nervous tissue by thin-layer chromatography in silica gel. Cholesterol and "minor" lipids were removed from the start by the first chromatography in the system ether-benzol-ethanol-acetic acid. Phospholipids and galactolipids, which remained at the start, were separated by the repeated chromatography in the system chloroform-methanol-NH4OH. The purified fractions of cholesterol, cerebrosides, sulfatides and the total fraction of phospholipids were obtained from 100-150 mg of nervous tissue in amounts, sufficient for estimation of their specific radioactivity in experiments with 14C.

Animals↗

3-O-acetyl-sphingosine-series myelin glycolipids: characterization of novel 3-O-acetyl-sphingosine galactosylceramide.

Several glycosphingolipids, less polar than galactosylceramide (GalCer), have been purified from rat brain and designated as fast migrating cerebrosides (FMCs). They co-appear with GalCer during myelinogenesis, reach a peak concentration at postnatal day 25-30 and are derivatives of GalCer. Extensive structural analysis of the partially methylated alditol acetates, mass-spectrometry, and (1)H- and (13)C-nuclear magnetic resonance (NMR) spectroscopy unequivocally established the structure of two of these FMCs as 3-O-acetyl-sphingosine GalCer with non-hydroxy and hydroxy fatty N-acylation respectively. That is, an acetyl group is linked at the C3-OH of the sphingosine base of GalCer. In addition, NMR spectroscopy of all of the purified FMCs indicates that they contain a 3-O-acetyl group linked with sphingosine and thus delineates a novel series. Several lines of evidence indicate that FMCs are myelin constituents. FMCs, enriched in both central nervous system (CNS) and peripheral nervous system (PNS) myelin, are concentrated in spinal cord and white matter that are composed of myelinated nerve fibers. There is N-acylation with alpha-hydroxy and C18 and C24 fatty acids, all characteristic of myelin components. They disappear along with GalCer in the murine genetic dysmyelinating disorders, jimpy and quaking, and in a knockout mutant which is devoid of GalCer. In addition, a decrease in FMC and GalCer concentration has been found in Krabbé's disease, a human genetic dysmyelinating disorder.

Animals↗