Abnormal Ganglioside Research Articles

Tricyclic antidepressant desipramine induces stereospecific opiate binding and lipid modifications in rat glioma C6 cells

Incubation for 48 hours of C6 glioma cell cultures with 10 −4M tricyclic antidepressant desipramine gave rise to a quantitative increase of total lipids and to qualitative modifications of glycosphingolipids involving detection by thin-layer chromatography of spots migrating according to cerebroside and sulfatide and presence of an abnormal ganglioside pattern. These lipid modifications were associated with the appearance of stereospecific binding of opiates (dihydromorphine) with a dissociation constant of 30–60 nM. These results favor an important role of lipids in opioid receptor function.

Differential Cellular Enrichment of Gangliosides in the Mouse Cerebellum: Analysis Using Neurological Mutants

Abstract: The cellular distribution of gangliosides in the cerebellum was studied in a series of adult mouse mutants that lose specific populations of neurons. The weaver (wv) mutation destroys the vast majority of granule cells, whereas the Purkinje cell degeneration mutation (pcd) destroys the vast majority of Purkinje cells. The staggerer (sg) and lurcher (Lc) mutations, on the other hand, destroy the vast majority of both granule and Purkinje cells. A proliferation of reactive glial cells, which occurs as a consequence of neuronal loss, has been reported in the sg/sg and pcd/pcd mutants, but not in the wv/wv mutant. Compared with the normal (+/+) mice, the concentration (μg/100 mg dry weight) of GD1a was significantly reduced in those mutants that lost granule cells, but was not reduced in the pcd/pcd mutant. The concentration of GTIa, on the other hand, was significantly reduced in those mutants that lost Purkinje cells, but was not reduced in the wv/wv mutant. A significant elevation in the concentration of GD3, which may be related to the proliferation of reactive glial cells, was observed in the pcd/pcd, sglsg, and Lc/+ mutants, but was not observed in the wv/wv mutant. Because these ganglioside abnormalities were confined to the cerebellum, they cannot result from genetic defects in ganglioside metabolism. Instead, these abnormalities result from a differential enrichment of gangliosides in neural membranes. Our findings suggest that GDT1a is more heavily concentrated in granule cells than Purkinje cells, whereas the opposite appears true for GTla. It also appears that GD3 is enriched in reactive glial cells and may play an important role during the morphological transformation of neural membranes.

Hormonal effects on the development of rat brain gangliosides—I. Cortisol

Ganglioside N-acetylneuraminic acid (NANA) content and distribution and that of incorporated radionuclide from 1-[ 14C]glucosamine were investigated under conditions of normal development and after administration of 500 μg of cortisol at 1 day of age. Cortisol delayed significantly the rise in tissue concentration of NANA usually occurring between 2 and 20 days of age. Cerebellum was most severely affected and brainstem least. The incorporation of radionuclide was depressed during the developmental lag then increased beyond controls from 6 to 10 days, and was again below that of controls in cerebrum and brainstem at 20 days. Individual study of seven chromatographically distinct resorcinol-positive compounds indicated that cortisol treatment favored accumulation and labeling of the more polar gangliosides initially, but resulted at 20 days of age in an abnormal ganglioside pattern with less polar compounds present disproportionately.

Abnormal adenylate cyclase activity and altered membrane gangliosides in thyroid cells from patients with graves disease

Summary Cultured thyroid cells from patients with Graves' disease have elevated cyclic AMP levels when grown in the presence of thyrotropin, and lowered levels in the absence of thyrotropin when compared to thyroid cells from normal individuals or patients with adenomas or colloidonodular goiters. This difference is correlated with an abnormal ganglioside pattern in the membranes from Graves' disease thyroids. These membranes are, however, indistinguishable in their binding of 125 I-thyrotropin. These findings are discussed as they relate to current views concerning the structural components of the thyrotropin receptor and the role of these components in the transmission of the thyrotropin message to the cell.

Oligosaccharide storage in brains from patients with fucosidosis, GM1-gangliosidosis and GM2-gangliosidosis (Sandhoff's disease).

Abstract— Analysis of whole autopsy brain from a patient with fucosidosis (α‐fucosidase deficiency) revealed minor storage of H‐antigen glycolipid [Fuc (α, 1→2) Gal‐GlcNAc‐Gal‐Glc‐Ceramide] and a slightly abnormal ganglioside composition in the form of a two‐fold elevation of GM1 and the presence of a fucose‐containing glycolipid (a minor component) which co‐migrated with GD1a. The major storage materials in fucosidosis brain were an oligosaccharide (Fuc‐Gal‐GlcNAc‐Man[Fuc‐Gal‐GlcNAc‐Man]‐ManGlcNAc) and a disaccharide [Fuc(α, 1→6)‐GlcNAc] in the approximate ratio of 5:1. Lesser amounts of a related oligosaccharide (Gal‐GlcNAc‐Man[Gal‐GlcNAc‐Man]‐Man‐GlcNAc) were isolated from the brain of patients with GM1‐gangliosidosis (Types I and II) where the major storage material is known to be GM1‐ganglioside (Gal (β, 1→3)GalNAc(β, 1→4) [NeuNAcf(α, 2→3) Gal(β, 1→4)Glc‐Ceramide). Similarly, a related oligosaccharide (GlcNAc‐Man [GlcNAc‐Man]‐Man‐GlcNAc) was isolated from the brain of a patient with a total deficiency of N‐acetyl‐β‐d‐hexosaminidase (Sandhoff variant of GM2‐gangliosidosis) where the major storage products are known to be GM2‐ganglioside (GalNAc (β 1→4) [NeuNAc (α, 2→3)Gal(β, 1→4)Glc‐Ceramine) and its asialo derivative. These studies indicate that glycoproteins containing at least 2 mol of l‐fucose per oligosaccharide unit are normally catabolized in human brain. Further, it appears that such glycoproteins are initially catabolized by an endo‐N‐acetylglucosaminidase to release an oligosaccharide which is then degraded by the sequential action of exo‐glycosidases.

Absence of brain ganglioside abnormalities in shambling mutant mice

Absence of brain ganglioside abnormalities in shambling mutant mice

Abnormal ganglioside accumulation in cultured fibroblasts from patients with mucolipidosis IV

Extracts of cultured skin fibroblasts derived from patients with mucolipidosis IV showed a marked increase and altered distribution of GM 3 and GD 3 gangliosides. GD 3 is elevated 1.5–2 times that of normal whereas GM 3 is elevated to a lesser extent. No abnormalities were found in the neutral glycolipids. These two gangliosides apparently comprise most of the accumulated lipid-like material observed on ultrastructural analysis in this disease.

The brain ganglioside pattern in presenile and senile dementia.

Ganglioside lipid fractions in the cerebral gray matter were studied in 13 patients with various forms of dementia. In 2 patients with Alzheimer's disease and 1 with senile dementia an abnormal ganglioside pattern was observed, i.e., a decrease of G1 and G3 in association with an increase of G2. In the light of the total clinical and histologic data on the patients, it would seem that this disturbance in ganglioside metabolism may be a phenomenon accompanying extensive degradation of brain tissue rather than a factor in the etiology of dementia.

SYNTHESIS OF AN ABNORMAL GANGLIOSIDE BY SKIN FIBROBLASTS IN SANDHOFF'S DISEASE

In Sandhoff's disease, following a complete β-hexosaminidase deficiency, there is an accumulation of at least 3 different glycolipids. Upon electron microscopy, cultured fibroblasts from a patient with this condition contained a small number of lamellar inclusions. Major glycolipids in normal confluent cultured fibroblasts are ceramide trihexoside and hematoside. In 2 fibroblast lines from unrelated children with Sandhoff's disease, we detected a ganglioside with an Rf similar to GM2-ganglioside. Complete characterization has not yet been completed. Both 14C-mannosamine and 14C-glucosamine were incorporated into this lipid which thus contains sialic acid and hexosamine. On the basis of this composition, this glycolipid may represent GM2-ganglioside. This compound has never been detected in normal or pathological control fibroblasts, including infantile Tay-Sachs disease, or the other sphingolipidoses. (Supported by the Dept. Mental Hygiene, State of California and PHS Grant HD-04612.)

GENERALIZED N-ACETYLNEURAMINIC STORAGE WITH SIALURIA

This case has remained unique (Fontaine, et al, Helv. Paed. Acta:23, Suppl. 17, 1968). The patient presented with mental retardation, seizures, mildly dysmorphic features, and sialuria, reaching several grams per day. The biochemical defect has not been identified. We studied biopsies from liver, kidney and skin. Hematoside level was normal in all these specimens as well as in erythrocytes. No abnormal ganglioside was detected. Significant increase of free N-acetylneuraminic acid was observed in liver, kidney and cultured skin fibroblasts. The ultrastructure of skin fibroblasts revealed scarce inclusions containing lamellar tubular cores, but no mitochondrial abnormalities such as were previously found in the liver. Skin fibroblasts were submitted to a loading test by incubation in a medium containing from 100 to 400 μg neuraminic acid per ml. This caused an increase in membrane-bound inclusions. When incubated with 14C-mannosamine, the precursor was normally incorporated into hematoside and glycoproteins. This suggested that these cells were capable of synthesizing CMP-neuraminic acid. A deficiency of neuraminic aldolase could not be disproven owing to an insufficient amount of available tissue. (Supported in part by PHS Grant HD-04612.)

Subacute sclerosing leucoencephalitis with an unusual pattern of cortical gangliosides. Chemical and electron microscopical study of a brain biopsy.

Brain biopsy was performed on a 7-year-old girl showing a typical clinical course of SSLE. Chemical analysis on cerebral cortex indicated an abnormal pattern of gangliosides, showing elevation of the usually minor, normal components, at the expense of the main ganglioside fractions. The total amount of gangliosides was normal. Electron microscopic study showed ‘nucleiform’ and crystaloid bodies within neuronal nuclei. It was assumed that both abnormal ganglioside pattern and the neuronal inclusions are related to the viral infection.

G M1-gangliosidosis (Type II): Studies on a fibroblast cell strain

A fibroblast cell strain was developed from the skin of a patient with the Type II form of G M1-gangliosidosis. β-Galactosidase activity at pH 5 with two synthetic substrates was about 3% of normal. Two abnormal gangliosides were identified: One, as expected, migrated like the G M1-ganglioside of brain; the other behaved like the G M2-ganglioside of brain. The composition and source of this G M2-like ganglioside is postulated. The major neutral glycolipid, tentatively identified as a trihexosylceramide, was present in greater amounts, and incorporated more radioactivity from 14C-galactose, than normal. A glycosaminoglycan fraction labeled with 14C-galactose and not precipitated by CPC was found in great excess. It's elution from a molecular sieve was very similar to that of a non-CPC-precipitable, undersulfated keratan-sulfate-like polysaccharide previously isolated from liver of a patient with Type I G M1-gangliosidosis. Spherical cytoplasmic inclusions stained metachromatically with toluidine blue after fixation in 30% acetic acid. Metachromasia was largely abolished after fixation in absolute methanol. It is probable that the accumulation of specific gangliosides and glycosaminoglycans in the mutant cell strain is related directly to its deficient pH 5 β-galactosidase activity. Limited studies on a cell strain derived from a patient with Type I G M1-gangliosidosis revealed no differences from the Type II strain with respect to the gangliosides stored or the frequency, intensity, and quality of cytoplasmic toluidine blue metachromasia.

Abnormal gangliosides in Tay-Sachs disease, Niemann-Pick's disease, and gargoylism

The molar ratios of N-acetyl neuraminic acid, hexose, hexosamine, and sphingosine have been determined for the abnormal ganglioside in Tay-Sachs disease that was previously detected as a fast-moving band in thin-layer chromatography, and in two abnormal fast-moving bands of gangliosides from the cortex and white matter of the brain in cases of gargoylism and Niemann-Pick's disease. The fastest-moving ganglioside band in these two conditions contains neither hexosamine nor glucose.