Changes In Gangliosides Research Articles

Changes in ganglioside composition of photoreceptors during postnatal maturation of the rat retina.

To examine at which stage the unusual ganglioside composition observed in adult retinal photoreceptor cells was established, and to see whether ganglioside changes could be correlated to distinct maturational events, quantitative and qualitative variations in gangliosides within pure sheets of photoreceptors during postnatal differentiation and aging of retina were studied. Retinas were separated into their component layers, (particularly photoreceptor layers uncontaminated by other neuronal types) by exploiting a technique of mechanical separation by vibratome. We extracted lipids from the cell membranes and analyzed the ganglioside composition by high performance thin layer chromatography. The data show that from the earliest recordable postnatal age (6 days) until late in life (18 months), photoreceptors contain low quantities of lipid-bound N-acetyl neuraminic acid and a simplified ganglioside profile compared to inner retinal neurons. Specific ganglioside changes occur within photoreceptor cells during postnatal maturation and aging, with downregulation of a-pathway GM1 and overlapping upregulation of b-pathway GD1b taking place during the period corresponding to outer segment formation, correlating with the onset of retinal function.

Ganglioside changes in brain in chronic relapsing experimental allergic encephalomyelitis induced in the Lewis rat.

Chronic relapsing experimental allergic encephalomyelitis (CREAE) was induced in Lewis rats by inoculation with guinea-pig myelin and complete Freund's adjuvant followed by treatment with low-dose cyclosporin A. Rats were sacrificed at different phases of the disease (just before the onset of clinical signs, during the first clinical episode of CREAE and during the first recovery). Gangliosides were extracted from the brain, analysed after purification by HPTLC fractionation and quantified densitometrically. An increase of GM1, the main rat myelin ganglioside, and a decrease of GT1b, suggested to play a role in mediating the interactions between oligodendroglia and axons, were observed during the development of the CREAE. These findings indicating significant ganglioside changes in CREAE give further support to the concept concerning the involvement of gangliosides in autoimmune demyelination.

Ganglioside spinal cord changes in chronic relapsing experimental allergic encephalomyelitis induced in the Lewis rats.

Chronic relapsing experimental allergic encephalomyelitis (CREAE) was induced in Lewis rats by inoculation with guinea-pig myelin and complete Freund's adjuvant followed by treatment with low-dose cyclosporin A. Rats were sacrified at different phases of the disease (just before the onset of clinical signs, during the first clinical episode of CREAE and during the first recovery). Gangliosides were extracted from the spinal cord, analysed after purification by two-dimensional chromatography and quantified densitometrically. An increase of GM 1, the main rat myelin ganglioside, and a decrease of GT1b, suggested to play a role in mediating the interactions between oligodendroglia and axons, were observed during the development of the CREAE. These findings indicating significant ganglioside changes in CREAE give further support to the concept concerning the involvement of gangliosides in autoimmune demyelination.

Ganglioside Alterations in the Central and Peripheral Nervous Systems of Patients with Creutzfeldt-Jakob Disease

We have examined the distribution and composition of gangliosides in central and peripheral nervous tissues from two patients with Creutzfeldt-Jakob disease (CJD). There were marked decreases in total ganglioside levels in CJD, with reductions in the cerebral cortex and cerebellum in the order of 20–30% and 50% of control values, respectively, though in spinal cord and sciatic nerve total gangliosides were not significantly altered. The percentage distribution of individual gangliosides was characterized by marked increases in GD3 and GD2, contrasting with severe decreases in GD1a, GD1b, GT1b and GQ1b; such changes were found throughout the patients' nervous tissues. An abnormal long-chain base composition was detected with the d20:1 component being decreased to less than 50% of control values, in the cerebrum, cerebellum and spinal cord. Changes in gangliosides occurred even in those tissues not severely affected neuropathogically. These ganglioside abnormalities are discussed in relationship to the pathogenesis of CJD.

Changes of gangliosides and sialic acid in human milk during lactation

To elucidate the possible biological function of gangliosides in human milk, the content and composition of gangliosides in human milk during lactation were examined. Gangliosides were rich in human milk at the earlier stages of lactation. Themajor ganglioside was GD3. The content of GD3 decreased, while that of GM3 increased in the later stages of lactation. GDla, and poiysialogangliosides were rich in the milk at 6 days of postpartum. Levels of sialic acid and sialidase, a ganglioside-hydrolyzing enzyme, were also studied. The content of sialic acid and sialidase activity were decreased in the later stages of lactation. As the results, GD3, GDIa, polysialogangliosides, sialic acids (lipid-bound and protein-bound), and sialidase were rich in human colostrum and these components may play a role in immune system.

Interleukin-3-associated expression of gangliosides in mouse myelogenous leukemia NFS60 cells introduced with interleukin-3 gene: expression of ganglioside GD1a and key involvement of CMP-NeuAc:lactosylceramide alpha 2-->3-sialyltransferase in GD1a expression.

Murine interleukin-3 (IL-3)-associated expression of gangliosides has been investigated using a gene transfection technique. A murine IL-3 cDNA was introduced into the parental NFS60-17 cells that was exclusively dependent on IL-3. We analyzed the glycosphingolipids from the parental cells and the transfected cells by fast atom bombardment mass spectrometry analyses and/or immunostaining techniques using specific antibodies. Two major gangliosides, IV3NeuAc-GgOse4Cer (GM1b) and IV3-NeuAc,III6NeuAc-GgOse4Cer (GD1 alpha), were expressed, in the parental cells. By contrast, in the IL-3 gene-transfected cells, a ganglioside IV3NeuAc,II3NeuAc-GgOse4Cer (GD1a) was strikingly expressed, in addition to GM1b and GD1 alpha that were already present in the parental cells. In spite of various IL-3-secreting capabilities, all transfectants investigated have exhibited the same ganglioside patterns and expressed GD1a. Furthermore, the appearance of GD1a was a consequence of the up-regulation of a single glycosyltransferase, CMP-NeuAc:lactosylceramide alpha 2-->3-sialytransferase (GM3 synthase). Activities of the other downstream glycosyltransferases that were involved in GD1a synthesis were not significantly different between the parental and the transfected cells. According to these data, the progression of tumor stage by the acquisition of autonomous cell growth ability after IL-3 gene transfection resulted in dramatic changes in cell surface gangliosides and their biosynthetic pathways. GD1a could be considered as an IL-3-associated ganglioside and was expressed in a tight connection with a single glycosyltransferase (GM3 synthase) up-regulation and with IL-3 expression in murine myelogenous leukemia cells.

Glycoprotein and ganglioside changes in human trophoblasts after exposure to pulsed Doppler ultrasound

Changes in glycoprotein and ganglioside composition in human trophoblasts (eighth week of gestation) after in vitro exposure to pulsed Doppler ultrasound (pulse duration 1.22 μs; repetition frequency 11.1 kHz; center frequency 4 MHz; ISPPA = 175.5 W/cm 2; ISPTA = 0.59 W/cm 2) were investigated. Evacuated trophoblasts were divided in two halves and insonated for 10 min on top of a 6-cm layer of 5% gelatin in 50-mL tubes (Falcon) at 37°C. One half of each trophoblast was sham insonated and served as an internal control. After insonation trophoblasts were maintained at 37°C for 24 h. Glycoproteins were detected using α- d-mannose specific lectins from Galanthus nivalis and Narcissus pseudonarcissus. A decrease in the expression of mannose containing glycoprotein mgp47 and an increase in expression of mgp54 were observed. Ganglioside composition was also significantly altered. Concentrations of two gangliosides migrating similarly to GM2, and one similarly to GQ1, decreased by more than 75%. At the same time, concentrations of one ganglioside migrating similarly to GM3, and two other unidetified gangliosides increased two- to fourfold.

Facile Methods for Isolation and Determination of Gangliosides in a Small Scale: Age-Related Changes of Gangliosides in Mouse Brain Synaptic Plasma Membranes

A quantitative isolation method for gangliosides from small sizes of samples has been developed. Total lipids were separated into gangliosides and other lipids using Phenyl Sepharose column chromatography. Gangliosides were recovered in a yield of 99%. Determination of gangliosides was performed by gas chromatography-mass spectrometry using a selected ion-monitoring technique. These combined methods can provide quantitative isolation and determination of gangliosides from as little as 10 mg fresh brain tissues or 0.5 mg protein of membrane fractions. The present methods were successfully applied to the analysis of gangliosides from mouse brain synaptic plasma membranes to reveal that the ganglioside contents and composition remain constant from adult to senescence.

Rat brain gangliosides during hypoxia

The content and composition of rat brain gangliosides in various forms of hypoxia is studied. The content of lipid-bound sialic acid is found to be considerably elevated in the earliest stages of hypoxia combined with hypercapnia and cooling. The content of gangliosides remains elevated over the hypoxia period and returns to control values 48 hours after its discontinuation. Repeated hypoxia results in the same changes in rat brain gangliosides. Hemic sodium nitrite-induced hypoxia also leads to a reliable elevation of these lipids. Hypoxia also causes changes in the content of individual ganglioside fractions: a decreased content of mono- and an increased content of polysialogangliosides.

Glycosphingolipids in cultured lens epithelial cells from dog and rhesus monkey.

Vertebrate lens tissues contain several species of acidic and neutral glycosphingolipids in relatively high amounts. However, the epithelia with capsule from dog and rhesus monkey lenses had a simpler composition and lower content of glycosphingolipids than whole lenses. Gangliosides and neutral glycosphingolipids in monolayer cultures of lens epithelial cells were also different from those in whole lenses. Although alpha-galactosyl (Gal alpha 1-3Gal-R) or Lewis(x) (Gal beta 1-4[Fuc alpha 1-3]GlcNAc-R) epitopes were found in glycosphingolipids from whole lenses, they were not detected in those from monolayer cultures of dog and rhesus monkey lens cells. In addition, significant changes in ganglio-series gangliosides were induced in monolayer cultures of both cells, where GM3 and GD3 were predominant. Immunofluorescence study revealed a characteristic distribution of cell surface gangliosides in confluent monolayers. These findings suggest that glycosphingolipid synthesis in lens epithelia is intrinsically different from that in cortical and nuclear fibres, and that the expression of Lewis(x) and alpha-galactosyl epitopes in glycosphingolipids appears to be associated with the differentiation of epithelial cells to fibres.

Altered ganglioside expression by SH-SY5Y cells upon retinoic acid-induced neuronal differentiation.

SH-SY5Y Neuroblastoma cells were used to study the effect of retinoic acid (RA)-induced differentiation on the expression of gangliosides and neuronal markers. In the presence of 10 microM RA, more than 70% of the cells differentiate to a neuronal phenotype within 8 days. They extend long neuritic processes and show an enhanced immuno-expression of neurone-specific enolase (NSE), neurofilament protein (NF-M), and polysialic acid (PSA). SH-SY5Y cells were found to express at least 12 different gangliosides. RA-induced neuronal differentiation led to a decrease in the content of GM2, GD3, and GD2 and to a 3-7 fold increased concentration of the ganglio-tetraosyl gangliosides GM1, GD1a, GT1a, GD1b, and GT1b. Thus, RA-induced neuronal differentiation of SH-SY5Y cells is accompanied by ganglioside changes similar to those observed during embryonic neuronal differentiation.

Nephron segment and cell-type specific expression of gangliosides in the developing and adult kidney

Despite the increasing knowledge of the role of gangliosides in normal and diseased tissues, little is known of the presence, distribution and functions of these molecules in the kidney. In this study we analyzed the main gangliosides of isolated glomeruli and cortical, medullary and papillary fractions of the human, rat and bovine kidneys biochemically. In addition, we used immunohistochemistry to visualize the distribution of GM1/GM2, GD2, GD3 and O-acetyl GD3 gangliosides along the nephron. Furthermore, we explored the species specific expression of gangliosides by comparing those from the rat, bovine and human kidney, and studied the pattern of ganglioside expression during development. In glomeruli, cortical tubuli, medullae and papillae, a relatively simple pattern of main gangliosides was observed as revealed by thin layer chromatographic (TLC) analysis in each species studied. Furthermore, considerable changes in the glomerular gangliosides during maturation were observed, with a complex type of gangliosides predominating during the fetal age and with a preference to more simple precursors upon maturation. Interestingly, the immunohistochemical detection revealed a distinct pattern of ganglioside compartmentation to various nephron segments or cell types. These findings provide a basis for studying the role of segment- and cell type-specific gangliosides for local functions.

Role of Gangliosides in Behavioral and Biochemical Actions of Alcohol: Cell Membrane Structure and Function

Alcohol exerts its pharmacological effects in adult brain by altering the physicochemical properties of cellular plasma membranes. Although alcohol does induce changes in membrane lipid composition, studies to relate these alterations to the development of behavioral tolerance to alcohol and the withdrawal effects have been unsuccessful. Actions of alcohol on developing brain are even more complex. Some of the reported effects include inhibition of embryogenesis, cell migration, and differentiation, including synaptogenesis. Gangliosides have neuroprotective action against a variety of neural insults (e.g., mechanical injury, drug toxicity, or hypoxic insult). This review addresses the role and significance of gangliosides in the CNS pathophysiology of alcohol exposure, as well as the effect of changes in endogenous gangliosides on membrane structure and function. We also describe the role of exogenous gangliosides in prevention of alcohol (acute and/or chronic)-induced CNS (prenatal and postnatal) neurotoxicity through their action on cellular plasma membranes. We propose that ganglioside's neuroprotective effects against alcohol neurotoxicity involve protection and restoration of plasma membrane structure (proteins and lipids) and thereby its function (ionic homeostasis, neurotransmitter receptor-mediated signal transduction). Thus gangliosides may have potential therapeutic use in treatment of alcohol-related problems.

Clinical heterogeneity of probable Alzheimer's disease.

When the symptomatology in probable Alzheimer's disease (AD) was studied, two subgroups emerged: one with predominant cortical symptoms of parietal type (AD type I) and another with general cognitive symptoms but absence of or only mild cortical symptoms (AD type II). In AD type I, the age at onset was significantly lower, confusional symptoms and leukoariosis on computerized tomographic scan were less frequent, and the ganglioside GM1 concentration in cerebrospinal fluid was significantly higher than in AD type II. Although the significance of changes in cerebrospinal fluid gangliosides has not been fully elucidated, it is possible that the increase in cerebrospinal fluid GM1 reflects a more severe degeneration of neurons and synapses in AD type I than in AD type II. The relation between symptomatology and leukoariosis suggests that leukoariosis has clinical significance in AD patients. The total results suggest heterogeneity of probable AD. AD type I appears to constitute a classic AD subgroup with memory disturbances and marked cortical symptoms of parietal type, while age-related changes, vascular changes, and leukoariosis may be responsible for the more generalized symptomatology in AD type II.

Lack of cell density-dependent changes in gangliosides of rat primary culture neurons.

Cell density-dependent changes in neuronal gangliosides, primarily relating to neurite outgrowth under dense to sparse conditions, were examined at cell seeding densities over an 8-fold range. During the first 24 h of incubation, the dissociated fetal rat neurons showed characteristic protrusion of neurites as a function of cell density. Ganglioside and protein contents per the same cell numbers were higher in dense cultures than sparse ones. However, the ganglioside pattern was essentially unchanged from dense to sparse culture, showing a predominance of GD3 and GT1b. The biosynthetic activity of gangliosides, as estimated by the incorporation of 3H-labeled N-acetyl-D-mannosamine, a precursor of sialic acid, was similar at various cell densities, with the labeling of b-series gangliosides predominating. The expression of neuronal gangliosides was monitored by indirect immunofluorescence using anti-GM1 antibody, but was found to be poor. A2B5 antigen, which was mainly identified as GT1b, appeared to be readily expressed on cell surfaces in sparse cultures. In contrast, the highly polysialylated form of the neural cell adhesion molecule (NCAM-H) was fully expressed on both the neurites and cell soma at various cell densities. The results suggest that the polysialic acids in NCAM have more important roles in neurite outgrowth than gangliosides, since the composition and synthesis of gangliosides are not affected by cell seeding density.

Developmental changes in gangliosides in cultured cerebellar granule neurons.

The content and composition of gangliosides in cultures enriched in granule neurones and in astrocytes from rat cerebellum (P6-8) showed marked differences: astrocytes contained less than 10% of the amount of granule neurones and the profile was dominated by simple gangliosides with lactosyl ceramide backbone, while gangliosides of the 'b' series, which constitute about 40% in nerve cells, were virtually undetectable. Granule cell maturation was accompanied by a 16-fold increase in the ganglioside content during the initial 8 days in a serum-supplemented medium (S+), reaching a plateau much earlier and at a higher level than observed in the cerebellum in vivo. Developmental changes were characterized, as in vivo, by a pronounced decrease in the GD3 proportion and an increase in the 'b' series of gangliosides. Compared with S+, adhesion among cells and fibres is different in a serum-free medium (S-), in which the rise in cellular ganglioside content was less (30%), but the developmental changes in ganglioside profile were similar. However, in cultures in S- only, GM3 was not detectable, while the distribution of GM1 and GD3 indicated that maturation is retarded relative to cells in S+. Surface exposure of gangliosides (studied by the periodate/[3H]borohydride method) was similar under the two culture conditions. There was an initial delay, especially in S-, in the insertion of gangliosides into the plasma membrane, while the labelling of GD3 (the dominant ganglioside of immature granule cells) was very low compared with all the other species throughout the whole cultivation time.

Effect of colchicine on ganglioside composition of rat primary culture neurons.

Developmental changes in gangliosides in the course of neurite outgrowth were examined in dissociated fetal rat cerebral neurons in culture. About a 2-fold increase in ganglioside levels was seen with the progression of neurite formation for up to 24 h in predominantly neuronal cultures. Ganglioside patterns appeared to be unchanged during the first 24 h, subsequently consisted of higher amounts of GD3 and b-series gangliosides (such as GD1b, GT1b, and GQ1b), and lower amounts of a-series gangliosides (GM1 and GD1a). Although the addition of colchicine to the cell growth medium inhibited neurite outgrowth in developing neurons, little if any differences in ganglioside patterns were found between control and colchicine-treated cells. Ganglioside levels decreased slightly in colchicine-treated cells in agreement with the decrease in cell attachment to culture dishes. Although colchicine treatment 8 h after plating caused complete retraction of formed neurites, the ganglioside level of the cells continued to increase during the following 16-hour incubation. Thus, the data suggest that ganglioside synthesis in differentiating neurons does not primarily accompany the expansion in cell surfaces due to neurite formation, and raises the possibility that a large proportion of gangliosides is retained in intracellular compartments.

Chapter 28: Gangliosides and neuronal-astrocytic interactions

Publisher Summary This chapter describes: (1) the content and composition of gangliosides in astrocytes; (2) the developmental expression of gangliosides in the cells; (3) quantitative and qualitative changes of gangliosides in pathological conditions affecting astrocytes; and (4) possible functions of gangliosides in astrocyte. Gangliosides are a family of sialic acid-containing glycosphingo lipids found primarily in the plasma membrane of virtually all vertebrate tissues and are particularly abundant in the nervous system. They are localized mainly in the outer surface of the plasma membranes and constitute part of theglycocalyx network surrounding the cell surface. Thus, they are assumed to be crucial in determining the properties and functions of cells. The occurrence of these molecules is cell-specific and is known to undergo developmentally regulated changes. These changes correlate well with their putative functions in cell to cell recognition, interaction and adhesion. Hence, they serve as excellent markers for monitoring cellular events in normal and pathological conditions.

Role of Myelin‐Associated Neuraminidase in the Ganglioside Metabolism of Rat Brain Myelin

The role of myelin-associated neuraminidase in ganglioside metabolism was examined using rats of ages ranging from 17 to 97 days. The neuraminidase activity directed toward the ganglioside GM3 in the total myelin fraction was high during the period of active myelination and, thereafter, decreased rapidly to the adult level. The ganglioside composition became simpler during development with an increasing amount of GM1 and decreasing percentages of di- and polysialogangliosides. The decrease in the proportion of GD1a was most prominent, whereas relative amounts of GD1b and GT1b increased transiently before reducing to the adult levels. The heavy myelin subfraction contained higher percentages of di- and polysialo-species compared to the light myelin fraction at young and adult ages. The in vitro incubation of myelin of young rats under an optimal condition for neuraminidase action produced a profile of ganglioside changes similar to that observed in in vivo development. These results strongly suggest that myelin-associated neuraminidase may play a pivotal role in the developmental changes in the ganglioside composition of rat brain myelin.

血液細胞および神経系組織における糖脂質の多様な役割

General review on the recent progress of glycosphingolipids was described both in various kinds of blood cells. and in the nervous tissues. First, the cell membrane glycosphingolipids were summarized and discussed as important markers for the maturation and differentiation from immature blood cells to mature myeloid and lymphoid cells, erythrocytes and platelets, as specific ligands to some selectins for cell-cell recognition and as roles of immunological responses.Second, the biological significance of myelin lipids, particularly cerebroside and sulfatide, changes of neuronal gangliosides in the development of neuronal cells, abnormal gangliosides in the brain tumors and biological activities of gangliosides were mentioned.