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PubMed1991

Developmental profiles of arylsulfatases A and B in rat cerebral cortex and spinal cord.

Arylsulfatases A (EC 3.1.6.1) and B (EC 3.1.6.12) are lysosomal enzymes that can remove sulfate groups from sulfatides and sulfo-glycosaminoglycans, respectively. The activities of these enzymes in cerebral cortex and in spinal cord of developing rat pups were measured. The tissues were homogenized and the arylsulfatases A and B in the soluble fraction were separated from each other by anion exchange chromatography on DE-52 cellulose. Subsequently, the enzyme activities were assayed with p-nitrocatechol sulfate as substrate at 37 degrees C and pH 5.6. We observed a developmental profile of arylsulfatase A, similar to that previously reported for cerebroside sulfatase (EC 3.1.6.8; (Van der Pal et al. (1990) Biochim. Biophys. Acta 1043, 91-96]. The activity of arylsulfatase A increased gradually during development, whereas arylsulfatase B rose more steeply, peaked around day 15 and declined thereafter. As a consequence the ratio between B and A forms of arylsulfatase dropped from about 4 in 1-week-old pups to 2.2 (cortex) and 0.7 (cord) in 7-week-old rat pups.

Animals↗
PubMed1986

Involvement of sulfatide in activation of protein kinase C by tumor promoters.

Sulfatide (cerebroside sulfate) activated protein kinase C to the same extent as phosphatidylserine did with the tumor promoters, 12-O-tetradecanoylphorbol-13-acetate (TPA), teleocidin and debromoaplysiatoxin. Sulfatide and phosphatidylserine both induced specific binding of [3H]TPA to protein kinase C, although the ratios of specific to non-specific [3H]TPA binding to protein kinase C with the two were not the same. It is concluded that sulfatide is involved in activation of protein kinase C by tumor promoters in a slightly different way from phosphatidylserine.

Animals↗
PubMed1984

Microheterogeneity of arylsulfatase a: Treatment with hydrolytic enzymes.

Cerebroside sulfatase also known as arylsulfatase A from human liver displays six microheteromer bands upon narrow pH range isoelectric focusing. Sialic acid residues only partially account for this enzyme multiplicity since neuraminidase treatment reduces the number of bands to three. Uptake studies with cultured fibroblasts strongly suggest arylsulfatase A has covalently bound mannose 6-phosphate residues. However, treatment with alkaline phosphatase and a battery of glycohydrolases failed to reduce the number of enzyme charge forms below three. These results imply that the neuraminidase-resistant charge microheterogeneity is not due to structures associated with the carbohydrate moiety of arylsulfatase A.

Cells, Cultured↗
PubMed1986

Expression of myelin proteins characteristic of fish and tetrapods by Polypterus revitalizes long discredited phylogenetic links.

CNS myelin constituents were used as evolutionary markers to study the controversial relationships of Polypterus with bony fishes, lungfishes and amphibians. The occurrence in Polypterus CNS of myelin proteins similar to those observed in the sterlet confirms its close relationship to Chondrostei. However, the simultaneous presence of proteolipid protein (PLP) demonstrates the existence of a long discredited relationship between Polypterus and tetrapods, and also with the lungfish Protopterus, an ally of tetrapods. As in the lungfish Protopterus, Polypterus cerebrosides and sulfatides contained alpha-hydroxy fatty acids. In contrast to Protopterus CNS myelin which carries glycosylated PLP but lacks Po-like component and the 36 kDa protein, Polypterus possesses these two bony fish myelin components. Furthermore, the presence of aglycosylated PLP in Polypterus, as in higher vertebrates, differentiates it from Protopterus. The simultaneous presence in Polypterus of myelin constituents from fish and land vertebrates indicates that Polypterids occupy an unusual intermediate phylogenetic position. The near absence of 2',3'-cyclic nucleotide 3'-phosphodiesterase activity in Polypterus. Protopterus, and in other fishes, confirmed by the lack of Wolfgram protein, establishes this myelin enzyme as the only CNS myelin constituent specifically expressed by tetrapods.

2',3'-Cyclic-Nucleotide Phosphodiesterases↗
PubMed1986

Regulation of galactocerebroside and sulfatide synthesis by hormones in chick neural cultures.

Chick neural cultures were used to study effects of insulin, thyrotropin releasing hormone, growth hormone and glucagon on myelin lipid synthesis in vitro. The incorporation of [3H]galactose into myelin associated lipids such as cerebroside and sulfatide was used as an index for various hormonal effects on myelination. The data suggest that these hormones were effective on myelin lipid synthesis only in the central nervous cells, not in the peripheral nerve cells.

Animals↗
PubMed1977

The dispersion of cholesterol with phospholipids and glycolipids.

Of the polar lipids studied (phospholipids and glycolipids), only phosphatidylcholine and sphingomyelin can disperse in water with up to 2 mol cholesterol/mol polar lipid. However, mixtures of phosphatidylethanolamine with small amounts of phosphatidylcholine and mixed lipids from mitochondria and myelin will also form sterol-rich dispersions. Steroids in which the 3beta-OH group is replaced by an oxo function do not form such steroid-rich dispersions. Electron microscopy and optical rotatory dispersion (ORD) show that sterols disperse with cerebrosides and gangliosides to form cylindrical structures with the regions around C atoms 3 and 7 of the sterol in less polar environments than those they occupy in phospholipid liposomes. It is proposed that choline-containing phospholipids facilitate entry of sterol molecules into the outer leaflet of cell surface membranes but that the phospholipid composition itself will not give rise to an asymmetric distribution of sterol in membranes with a high cholesterol content.

Cerebrosides↗
PubMed1982

Compositional aspects of lipid hydration.

The effect of various lipids such as cerebrosides, gangliosides and dipalmitoyl lecithin (DPPC) on the lowering of the melting temperature of water, was determined by differential scanning calorimetry (DSC). The lowering of the melting temperature, and the number of water molecules per lipid molecule which apparently do not undergo melting, increase with the increase of the size and charge of the polar group and with the unsaturation of the hydrocarbon chains. Freezing curves show supercooling to about -20 degrees C. It was found that the number of apparently unfreezable water molecules is about four for glucocerebroside from Gaucher's spleen and about eight or nine for galactocerebroside from bovine brain. In gangliosides from bovine brain the following number of water molecules/lipid molecule are apparently unfreezable: 22-30 in GM1, 33-40 in GD1a + GD1b while a fraction of gangliosides containing 75% GQ1b and 25% GT1b affects up to 60 molecules of water/molecule of lipid. A zwitterionic DPPC molecule removes apparently six to seven water molecules from freezing. There is no indication that the apparently unfreezable water molecules are in a distinct state. It is suggested that they freeze at very low temperatures producing a flat tail preceding the transition peak which cannot be discerned from the base line.

Animals↗
PubMed1991

Arylsulfatases A and B in EBV-transformed lymphoid cell lines: studies on their molecular forms in cells from patients with inborn sulfatase deficiencies. Comparative diagnostic value of enzymatic assays.

The enzyme activity of arylsulfatase A and arylsulfatase B was studied in Epstein-Barr virus-transformed lymphoid cell lines established from control individuals and patients affected with metachromatic leukodystrophy, mucopolysaccharidosis type VI (or Maroteaux-Lamy syndrome) and multiple sulfatase deficiency. Lymphoid cells derived from patients with metachromatic leukodystrophy showed a severe deficiency in cerebroside sulfatase activity, as measured using radiolabelled sulfatide, but some residual activity of arylsulfatase A when measured with the chromogenic substrate, para-nitrocatechol sulfate. Lymphoid cells from mucopolysaccharidosis type VI had virtually no arylsulfatase B activity. In cells from patients with multiple sulfatase deficiency, the activities of lysosomal sulfatases as well as steroid sulfatase were deficient. Study of the molecular forms of arylsulfatases confirmed the complete deficiency of arylsulfatase A and arylsulfatase B activities in metachromatic leukodystrophy and mucopolysaccharidosis type VI lymphoid cells, respectively. The arylsulfatase A defect in metachromatic leukodys-lymphoid cells, respectively. The arylsulfatase A defect in metachromatic leukodystrophy cells could be demonstrated on focused fractions even using the artificial substrates, para-nitrocatechol sulfate and 4-methylumbelliferyl sulfate. To investigate the discrepancy of the arylsulfatase A activity data observed between whole cell homogenates and focused fractions when using the synthetic substrates, assays were tentatively performed for optimizing the determination of arylsulfatase A on crude homogenates of lymphoid cells. Although this work has indicated methodological limitations of the enzymatic assay of arylsulfatase A in lymphoid cells using methylumbelliferyl sulfate, it emphasizes the validity of lymphoid cell lines as an experimental model for the study of inborn deficiencies of arylsulfatases A and B.

B-Lymphocytes↗
PubMed1983

Growth properties and biochemical characterization of mouse Schwann cells cultured in vitro.

Purified secondary cultures of mouse Schwann cells (less than 5% fibroblast contamination) have been obtained by taking advantage of the differential adhesion of Schwann cells and fibroblasts during trypsinization. The growth properties of the purified subcultures changed with time in culture. Cells passaged after 5 days in vitro (DIV) divided rapidly (doubling time 22 h), whereas cells that had been in vitro for longer periods progressively decreased their growth rate, becoming quiescent after 20 or more days. Schwann cells lacked the Thy 1.2 surface antigen, but were positively stained with antigalactocerebroside antibodies after prefixation. Biochemical analyses showed Schwann cells to be enriched in the activities of enzymes characteristic of the myelin-forming cells: 2'3'-cyclic nucleotide 3'-phosphodiesterase (CNP), cerebroside sulfotransferase (CST) and UDP-galactose: ceramide galactosyltransferase (CGalT).

2',3'-Cyclic Nucleotide 3'-Phosphodiesterase↗
PubMed1980

A new assay for lytic anti-galactocerebroside (GC) antibodies employing 56Rubidium release from GC-labelled liposomes.

This assay is a quick and sensitive method for determination of antibodies to galactocerebroside (GC). The use of antibody-plus-complement radioisotope release from GC-bearing liposomes allows numerous samples to be handled in semi-automated fashion. By substituting alternative gangliosides or cerebrosides, the technique may be simply modified for assay of antibodies to other substances.

Animals↗
PubMed1984

A micromethod for absorption of specific antibody using an enzyme-linked immunosorbent assay (ELISA).

A procedure for absorbing specific antibodies using an enzyme-linked immunosorbent assay system (ELISA) has been developed. This is accomplished by serial absorption of the serum using less than 20 micrograms antigen. The sera can then be used in an ELISA system to test reactivity with other antigens. The system was tested by absorbing anti-myelin or anti-cerebroside antibodies and comparing these results with bulk absorption.

Absorption↗
PubMed1993

Antibodies specific for the lipid-bound form of myelin basic protein during experimental autoimmune encephalomyelitis.

On the hypothesis that myelin basic protein isolated with surrounding lipids may constitute an autoantigen in demyelinating diseases, we studied the antibody response to the lipid-free and lipid-bound form of myelin basic protein during the course of experimental autoimmune encephalomyelitis induced in rats with either form of protein. Immunization with the lipid-bound form of myelin basic protein induced high titres of antibodies directed to the protein, accompanied by no antibodies to cerebroside 30 days after immunization. Antibodies specifically directed to the lipid-bound form of myelin basic protein were revealed after removal of antibodies recognizing the delipidated myelin basic protein. Anti lipid-bound myelin basic protein antibodies could already be detected at day 10 post-immunization, reaching a maximum at day 20 post-immunization. Demonstrations of antibodies entirely specific for the lipid-bound form of myelin basic protein suggests that this molecule may present epitopes not to be found in its already extensively studied primary structure, possibly the result of conformational changes following lipid binding.

Animals↗
PubMed1989

A fluorometric study of the interaction of bradykinin with lipids.

The interaction of bradykinin (BK) with lipids has been followed by steady-state fluorescence measurements. Addition of either cerebroside sulfate (CS) or phosphatidylinositol (PI), solubilized with the nonionic surfactant C12E8, to BK or its analogue [Gly6]-BK enhances the relative fluorescence intensity of peptide emission at 288 nm. Fluorometric titration of the peptide with lipid has been used to quantitate the interactions in terms of stoichiometry and equilibrium constant. Job's method of continuous variation for the BK-CS interaction gave a stoichiometry of 1:2 for the complex. The value of the equilibrium constant, K, for the interaction of either BK or [Gly6]-BK with CS is 1.5.10(4) M-1. The BK-PI interaction is weaker; K = 5.0.10(3) M-1. Although electrostatic forces no doubt play a major role in these interactions, measurements on the model peptide Gly-Phe-Gly indicate that the phenylalanine residues of BK are disposed in the hydrophobic environment provided by the lipid-C12E8 mixed micelle. 13C-NMR measurements on [99% 13C alpha-Gly6]-BK show that there is no change in its cis/trans ratio upon interaction with CS. The increase in the relative fluorescence intensity of BK accompanying its cooperative interaction with sodium dodecyl sulfate (SDS) implicates the role of hydrophobic forces in this interaction as well. These results bear on the interpretation of the changes in circular dichroism (CD) of BK caused by SDS.

Amino Acid Sequence↗
PubMed1985

Strain differences and laminar localization of structural neurochemical changes in aging rat.

To obtain comparisons of age-related microchemical changes in cerebral cortex of two commonly employed rat strains (Fischer 344 and Sprague-Dawley), neurochemical assays of substances regarded as quantitative indices of structural entities in brain were performed. These included DNA as a marker for cells, lipid sialoganglioside as an index of neuronal membrane mass, and galactocerebroside as an index of myelin. Fischer 344 rats were studied at 3-4 months (young), 14-16 months (middle age) and 25-28 months (old). Sprague-Dawleys were examined at 3-6 months (young), 14-17 months (middle age) and 25-28 months (old). Significant differences in the time courses of changes occurred; Fischer rats increased their brain weight at each aging point, while Sprague-Dawley rats reached stable brain weights by 4 months of age. Neither strain had a significant change in cell packing density of somatosensory cortex as measured by DNA. However, total ganglioside sialic acid declined in both strains, occurring by middle age in the Fischer and not until senescence in the Sprague-Dawley cortex. Cerebroside galactose increased in the Fischer between young and middle age, and was not further elevated in the older group. The Sprague-Dawley had its major increase in this marker between the middle aged and senescent groups. Intralaminar assays of these same markers in young and old Fisher 344 rats again indicated that DNA did not change, and that sialoganglioside was lost from all layers of the cortex in equal amounts. However, the increase in galactocerebroside resulted entirely from increases in the lower lamina of somatosensory cortex (lamina IV and below), suggesting on-going myelination of afferent and efferent axons. The time course of lipid membrane alteration is strain-dependent and selective as to cortical laminar localization. The findings are discussed in reference to human aging change in the same neurochemical indices.

Aging↗
PubMed1980

Neuritogenic activity of peripheral nerve myelin proteins in Lewis rats.

The neuritogenic activity of human peripheral nerve myelin proteins in Lewis rats was examined. Experimental allergic neuritis (EAN) was induced by human myelin fraction in all Lewis rats examined. Purified human P2 protein induced mild but definite EAN. None of purified P0 protein, total myelin lipids, gangliosides and cerebrosides induced EAN. Addition of total myelin lipids or gangliosides to the P2 protein did not enhance the neuritogenicity of the P2 protein. Bovine P2 protein, which was inactive in rabbits and guinea pigs, showed considerable neuritogenicity in Lewis rats. We postulate that induction of EAN depends both on the species of host animal and the source of antigen.

Animals↗
PubMed1984

Sumithion induced neurotoxicity in pigeons: effect on lipid metabolism of spinal cord.

Sumithion, a well known organophosphorus pesticide, produces hind leg paralysis and ataxia in pigeons at a dose of 5 mg/kg for 5 days. Histochemical changes in the spinal cord showed demyelination in the anterior portion of the thoracic region. Decreases in cholesterol, cerebroside and sulphatide and increases in cholesterol ester suggest demyelination in pigeons as a consequence of the neurotoxic effect of sumithion.

Animals↗
PubMed1979

Lipids from nerve tissues of the horseshoe crab, Limulus polyphemus.

1. The predominant lipids of nerve cords, ganglion and brain from horseshoe crabs were cholesterol (11% of lipid) and phospholipid (81% of lipid). 2. Major phospholipids were phosphatidyl ethanolamine and phosphatidyl choline with lesser amounts of phosphatidyl serine and phosphatidyl inositol and sphingomyelin. 3. The phospholipid fraction was characterized by a high content of plasmalogen, i.e. alk-1-enyl acyl phosphatides, so that 42% of the ethanolamine phosphatides were the plasmalogen, phosphatidal ethanolamine. 4. Phosphatidyl choline and phosphatidyl ethanolamine were high in polyunsaturation with 20:4 and 20:5 major fatty acids. Sphingomyelin had predominantly long chain saturated fatty acids. 5. Cerebrosides and gangliosides, which are associated with vertebrate nerve tissues, were absent from nerves of horseshoe crabs.

Animals↗
PubMed1996

High-performance liquid chromatography method with light-scattering detection for measurements of lipid class composition: analysis of brains from alcoholics.

A high-performance liquid chromatographic method with evaporative light-scattering detection was developed for the analysis of intact lipid classes in nervous tissue. The method had the ability to resolve plasmalogen-phosphatidyl-ethanolamine and diacyl-phosphatidylethanolamine along with other major phospholipid classes in a single run. This technique was employed for the investigation of the effects of chronic alcohol consumption on the membrane lipid class composition of human brains (alcoholics, n = 13; controls, n = 11). Measurements were performed on cholesterol, cerebrosides, sulfatides, phospholipids and sphingolipids in total lipid extracts of white matter, gray matter and cerebellar regions of human brains. No significant differences in the lipid class composition between the groups were observed.

Alcoholism↗