g(m1)-ganglioside and Gangliosidoses

Topics: alpha-Galactosidase; Arylsulfatases; beta-Galactosidase; Cystine; Fabry Disease; G(M1) Ganglioside; G(M2) Ganglioside; Galactosylceramidase; Gangliosidoses; Genetic Carrier Screening; Glycoproteins; Heparitin Sulfate; Humans; Hydrolases; Isoelectric Focusing; Isoenzymes; Kinetics; Leukodystrophy, Globoid Cell; Leukodystrophy, Metachromatic; Lipid Metabolism, Inborn Errors; Lysosomes; Metabolism, Inborn Errors; Molecular Weight; Mucolipidoses; Niemann-Pick Diseases; Sphingolipidoses; Sphingomyelin Phosphodiesterase

Topics: Animals; Cattle; Enzyme Activation; G(M1) Ganglioside; G(M2) Ganglioside; Gangliosidoses; Glucosylceramidase; Glycolipids; Glycosphingolipids; Hexosaminidases; Humans; Leukodystrophy, Metachromatic; Lysosomes; Mice; Protein Binding; Proteins; Rats; Substrate Specificity; Sulfatases

Recent years have witnessed considerable change in the conceptualization of the pathophysiology of the cognitive impairments in dementing disorders, as a result of synaptic neurochemical analyses. Profound reductions in the forebrain cholinergic projections occur in Alzheimer's disease. In GM1 gangliosidosis, variable alterations in neurotransmitter related processes that are located in synaptic membranes have been described. Exploitation of animal models of human disorders resulting in dementia may further clarify the dynamic alterations in the biochemical processes required for effective neurotransmission in cortex.

Topics: Alzheimer Disease; Animals; Brain; Cats; Cerebral Cortex; Cholinergic Fibers; Dementia; Disease Models, Animal; Dopamine; G(M1) Ganglioside; gamma-Aminobutyric Acid; Gangliosidoses; Glutamates; Glutamic Acid; Humans; Neurotransmitter Agents; Norepinephrine; Rats; Serotonin; Synaptic Transmission

Topics: Aspartylglucosaminuria; Carbohydrate Metabolism, Inborn Errors; Chromatography, Paper; Fucose; G(M1) Ganglioside; Gangliosidoses; Glucosidases; Glycoproteins; Humans; Lactose Intolerance; Mannose; Mucolipidoses; Mucopolysaccharidoses; Neuraminidase; Oligosaccharides; Sandhoff Disease

Topics: Adolescent; Adult; Animals; beta-Galactosidase; Cats; Cerebrovascular Disorders; Child, Preschool; Coronary Disease; Fabry Disease; Female; G(M1) Ganglioside; G(M2) Ganglioside; Gangliosidoses; Gaucher Disease; Glycolipids; Hexosaminidases; Humans; Infant; Infant, Newborn; Leukodystrophy, Globoid Cell; Leukodystrophy, Metachromatic; Male; Sodium-Potassium-Exchanging ATPase; Vascular Resistance

Topics: Animals; beta-Galactosidase; Child; Child, Preschool; Female; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Humans; Infant; Infant, Newborn; Isoenzymes; Liver; Male; Pregnancy; Species Specificity; Tissue Distribution

Topics: Adolescent; beta-Galactosidase; Brain Diseases, Metabolic; Child; Child, Preschool; Clinical Enzyme Tests; Female; G(M1) Ganglioside; G(M2) Ganglioside; Gangliosidoses; Humans; Infant; Infant, Newborn; Isoenzymes; Mass Screening; Metabolism, Inborn Errors; Models, Genetic; Phenotype; Pregnancy; Prenatal Diagnosis; Sandhoff Disease; Tay-Sachs Disease

Topics: G(M1) Ganglioside; G(M2) Ganglioside; G(M3) Ganglioside; Gangliosidoses; Genetic Carrier Screening; Genetic Counseling; Glycoside Hydrolases; Humans; Leukodystrophy, Metachromatic; Lipidoses; Mucopolysaccharidoses; Mucopolysaccharidosis I; Mucopolysaccharidosis III; Mucopolysaccharidosis IV; Mucopolysaccharidosis VI; Phenotype; Polymorphism, Genetic

Topics: Brain; G(M1) Ganglioside; G(M2) Ganglioside; Gangliosides; Gangliosidoses; Humans; Lipidoses

This article describes a case which seek medical advice for 2 months due to retrogressive development, The discovery of the characteristic fundus of the macular cherry-red spot is a key clue for further genetic analysis, GLB compound heterozygous mutations is detected, and enzymology results show that the acid B-galactose glucoside enzyme significantly decrease, fundus inspection help diagnosis GM1 gangliosidoses.. 患儿男性，11个月。生后5个月出现发育倒退，随后出现四肢无力和对响声的异常惊吓反应，求医2个月诊断不明，转诊至首都医科大学附属北京儿童医院，发现患儿双眼眼底黄斑樱桃红斑，以此为线索进一步基因分析显示GLB基因出现复合杂合子突变，酶学检查结果显示酸性β-半乳糖苷酶明显低于正常值，确诊GM1型神经节苷脂沉积症。.

Topics: G(M1) Ganglioside; Galactose; Gangliosidoses; Glucosides; Humans; Mutation

GM1 and GM2 gangliosidoses are progressive neurodegenerative lysosomal storage diseases resulting from the excessive accumulation of GM1 and GM2 gangliosides in the lysosomes, respectively. The diagnosis of gangliosidosis is carried out based on comprehensive findings using various types of specimens for histological, ultrastructural, biochemical and genetic analyses. Therefore, the partial absence or lack of specimens might have resulted in many undiagnosed cases. The aim of the present study was to establish immunohistochemical and immunofluorescent techniques for the auxiliary diagnosis of canine and feline gangliosidoses, using paraffin-embedded brain specimens stored for a long period.. Using hematoxylin and eosin staining, cytoplasmic accumulation of pale to eosinophilic granular materials in swollen neurons was observed in animals previously diagnosed with GM1 or GM2 gangliosidosis. The immunohistochemical and immunofluorescent techniques developed in this study clearly demonstrated the accumulated material to be either GM1 or GM2 ganglioside.. Immunohistochemical and immunofluorescent techniques using stored paraffin-embedded brain specimens are useful for the retrospective diagnosis of GM1 and GM2 gangliosidoses in dogs and cats.

Topics: Animals; Brain; Cat Diseases; Cats; Dog Diseases; Dogs; Fluorescent Antibody Technique; G(M1) Ganglioside; G(M2) Ganglioside; Gangliosidoses; Immunohistochemistry; Paraffin Embedding; Reproducibility of Results; Retrospective Studies

The ganglioside-activator protein is an essential cofactor for the lysosomal degradation of ganglioside GM2 (GM2) by beta-hexosaminidase A. It mediates the interaction between the water-soluble exohydrolase and its membrane-embedded glycolipid substrate at the lipid-water interphase. Mutations in the gene encoding this glycoprotein result in a fatal neurological storage disorder, the AB variant of GM2-gangliosidosis. In order to efficiently and sensitively probe the glycolipid binding and membrane activity of this cofactor, we synthesized two new fluorescent glycosphingolipid (GSL) probes, 2-NBD-GM1 and 2-NBD-GM2. Both compounds were synthesized in a convergent and multistep synthesis starting from the respective gangliosides isolated from natural sources. The added functionality of 2-aminogangliosides allowed us to introduce the chromophore into the region between the polar head group and the hydrophobic anchor of the lipid. Both fluorescent glycolipids exhibited an extremely low off-rate in model membranes and displayed very efficient resonance energy transfer to rhodamine-dioleoyl phosphoglycerol ethanolamine (rhodamine-PE) as acceptor. The binding to GM2-activator protein (GM2AP) and the degrading enzyme was shown to be unaltered compared to their natural analogues. A novel fluorescence-resonance energy transfer (FRET) assay was developed to monitor in real time the protein-mediated intervesicular transfer of these lipids from donor to acceptor liposomes. The data obtained indicate that this rapid and robust system presented here should serve as a valuable tool to probe quantitatively and comprehensively the membrane activity of GM2AP and other sphingolipid activator proteins and facilitate further structure-function studies aimed at delineating independently the lipid- and the enzyme-binding mode of these essential cofactors.

Topics: Animals; beta-N-Acetylhexosaminidases; Brain; Carbohydrate Sequence; Catalysis; Cattle; Chromatography, Thin Layer; Fluorescence Resonance Energy Transfer; Fluorescent Dyes; G(M1) Ganglioside; G(M2) Activator Protein; G(M2) Ganglioside; Gangliosides; Gangliosidoses; Glycolipids; Glycoproteins; Humans; Lipids; Models, Chemical; Molecular Sequence Data; Mutation; Spectrometry, Fluorescence; Sphingolipid Activator Proteins; Sphingolipids; Structure-Activity Relationship; Tay-Sachs Disease; Time Factors

A novel fluorescent ganglioside, sulforhodamine-GM1 was administered into cells derived from carriers and patients with different subtypes of GM2 gangliosidosis, resulting from various mutations in the gene encoding the lysosomal enzyme hexosaminidase (Hex) A. The cells used were skin fibroblasts and white blood cells, i.e. lymphocytes, monocytes and macrophages. In the severe infantile form of the GM2 gangliosidosis, Tay-Sachs disease, the sulforhodamine-GM1 was hydrolyzed within the lysosomes to the corresponding sulforhodamine-GM2 which, because of lack of Hex A activity, was not further degraded. In comparison, in the cells derived from GM2 gangliosidoses carriers, as well as pseudodeficient and adult forms of GM2 gangliosidosis, the sulforhodamine-GM2 was further processed and sequentially degraded by the lysosomal glycosidases to sulforhodamine-ceramide. The latter was converted to sulforhodamine-sphingomyelin, which was secreted into the culture medium. The fluorescence of the sulforhodamine ceramide in cell extracts and/or sulforhodamine-sphingomyelin in the culture medium was quantified and related to parallel data obtained using cells of normal individuals. This permitted distinguishing between the various GM2 gangliosidoses subtypes and relating the intracellular hydrolysis of sulforhodamine-GM1 to the genotypes of the respective GM2 gangliosidoses variants.

Topics: Cell Line; Fibroblasts; Fluorescence; G(M1) Ganglioside; Gangliosidoses; Humans; Leukocytes; Mutation; Rhodamines; Tay-Sachs Disease

GM1- and GM2-gangliosides were isolated from brain and radiolabelled. The labelled moieties were localized by hydrolysis with lysosomal enzymes, followed by thin-layer chromatography of the products. High-resolution loading tests with labelled gangliosides were developed and found to differentiate infantile and juvenile forms of GM1- and GM2-gangliosidoses as well as the identification of B, O and AB types of GM2-gangliosidosis.

Topics: Animals; Brain Chemistry; Chromatography, Thin Layer; Diagnosis, Differential; Fibroblasts; G(M1) Ganglioside; G(M2) Ganglioside; Gangliosidoses; Gangliosidosis, GM1; Genetic Variation; Humans; Kinetics; Mice; Sandhoff Disease; Tay-Sachs Disease

The clinical, morphologic, histochemical, and biochemical features of GM1-gangliosidosis in two canine models, English Springer Spaniel (ESS) and Portuguese Water Dog (PWD), have been compared. The disease onset, its clinical course, and survival period of the affected dogs were similar in both models. Skeletal dysplasia was noted radiographically at 2 months of age, whereas at 4 1/2 months of age there was progressive neurologic impairment. However, dwarfism and coarse facial features were seen only in ESS. Both models had similar deficiency in activity of lysosomal beta-galactosidase, but possessed a normal protein activator for GM1-beta-galactosidase. Both models stored GM1-ganglioside, asialo-GM1, and oligosaccharides in brain. Furthermore, only the PWD stored glycoproteins containing polylactosaminoglycans in visceral organs, and neither model stored them in the brain. Morphologically, both models demonstrated similar storage material in multiple tissues and cell types. The ultrastructure of the storage material was cell-type specific and identical in both models. However, some differences in the lectin staining pattern were noted. Our clinical, biochemical, and histochemical findings indicate that PWD and ESS may represent two different mutations of the beta-galactosidase gene. Moreover, the authors conclude that it is difficult, and inappropriate, to apply the human classification of GM1-gangliosidosis (i.e. infantile, juvenile, and adult forms) to these canine models.

Topics: Amniotic Fluid; Animals; Carbohydrate Metabolism; Disease Models, Animal; Dogs; G(M1) Ganglioside; Gangliosidoses; Lipid Metabolism; Microscopy, Electron; Placenta; Umbilical Cord

Phosphoinositide-specific phospholipase C and adenylyl cyclase were studied in brain cortical membranes from cats with GM1 and GM2 gangliosidosis. In contrast to brain cortical membranes from unaffected control cats, phospholipase C acting against exogenously supplied phosphoinositide substrates did not respond to stimulation by GTP gamma S, carbachol or fluoroaluminate in cortical membranes of cats with gangliosidosis. However, the enzyme was activated by calcium in membranes from affected cats to the same extent as in membranes from control cats. Basal adenylyl cyclase activity was increased 3-fold in cortical membranes of cats with GM1 and GM2 gangliosidosis, compared with unaffected sibling controls. Fluoroaluminate was equally effective in stimulating adenylyl cyclase in controls and in membranes of affected and normal cats. In addition, GppNHp was able to inhibit the forskolin-activated enzyme both in membranes from cats with gangliosidosis and sibling controls. These data suggest that the activation of phosphoinositide-specific phospholipase C in brain membranes by guanine nucleotide binding proteins is markedly impaired in GM1 and GM2 gangliosidoses.

Topics: Adenylyl Cyclases; Aluminum; Aluminum Chloride; Aluminum Compounds; Animals; Calcium; Carbachol; Cat Diseases; Cats; Cell Membrane; Cerebral Cortex; Chlorides; Colforsin; G(M1) Ganglioside; Gangliosidoses; Gangliosidosis, GM1; Guanosine 5'-O-(3-Thiotriphosphate); Guanylyl Imidodiphosphate; Kinetics; Phosphatidylinositol 4,5-Diphosphate; Phosphatidylinositol Diacylglycerol-Lyase; Phosphatidylinositols; Phosphoric Diester Hydrolases; Reference Values; Sodium Fluoride

Two new oligosaccharides were isolated from the urine of a patient with GM1 gangliosidosis. Final purification of the oligosaccharides was accomplished by capillary supercritical fluid chromatography. Structural analysis was by chemical analysis, chemical-ionization mass spectrometry and 400-MHz 1H-NMR spectroscopy, leading to two primary structures. The first is derived from a classical triantennary N-acetyllactosamine-type glycan: Gal beta 1-4GlcNAc beta 1-4(Gal beta 1-4GlcNAc beta 1-2)Man alpha 1-3Man beta 1-4GlcNAc. The second is unusual with a terminal disaccharide Gal beta 1-6Gal, which had not yet been described for glycans of the N-acetyllactosamine type: Gal beta 1-6Gal beta 1-4GlcNAc beta 1-2Man alpha 1-6Man beta 1-4GlcNAc.

Topics: Carbohydrate Conformation; Carbohydrate Sequence; Chromatography, Paper; G(M1) Ganglioside; Gangliosidoses; Glycosylation; Humans; Magnetic Resonance Spectroscopy; Mass Spectrometry; Molecular Sequence Data; Oligosaccharides; Proteoglycans

The pathogenesis of neuronal dysfunction in the gangliosidoses is poorly understood. Studies of the feline gangliosidoses and in vitro experiments implicate ganglioside inhibition of protein kinase C (PKC) in the pathogenesis of these neurological diseases. Therefore, in the present study, the binding of [3H]phorbol-12, 13 dibutyrate was measured to determine the levels of PKC in cerebral cortex of cats with GM1 gangliosidosis (mutant) and age matched normal siblings. This binding of ([3H]PDB) to cerebral cortex homogenates in both normal and mutant cats was highly specific. The specificity of receptors was ascertained also from displacement studies using nonradioactive phorbol ester analogues to displace [3H]PDB bound to its receptors. In both mutant and normal cat brain, phorbol 12,13-dibutyrate (PDB), 4-beta-phorbol 12,13-didecanoate (beta-PDD) and 4-beta-phorbol 12,13-dibenzoate (beta-PDBz) were highly potent (approximately to same degree) and effective in displacing [3H]PDB. On the other hand, 4-beta phorbol 12,13-diacetate (beta-PDA) was a weak displacer and 4-alpha-phorbol did not displace the bound [3H]PDB in either normal or mutant brain. Scatchard analysis of the binding data indicated a homogenous single class of binding sites in normal and mutant brain (Normal: Kd = 1.42 x 10(-7) M, Bmax = 8.40 pmoles/mg protein. Mutant: Kd = 1.60 x 10(-7) M, Bmax = 10.00 pmoles/mg protein). Sphingosine inhibited the binding to approximately the same extent in normal and mutant cortex. These studies demonstrate the presence of highly specific, homogenous, single type phorbol ester receptors in cerebral cortex of cats with GM1 gangliosidosis which are qualitatively and quantitatively similar to normal cat brain.

Topics: Animals; Binding, Competitive; Caenorhabditis elegans Proteins; Carrier Proteins; Cats; Cerebral Cortex; G(M1) Ganglioside; Gangliosidoses; Phorbol 12,13-Dibutyrate; Phorbol Esters; Protein Kinase C; Receptors, Drug; Sphingosine

Molecular analysis of the human beta-galactosidase gene revealed six different mutations in 10 of 11 Japanese GM1-gangliosidosis patients. They were the only abnormalities in each allele examined in this study. A 165-nucleotide duplication (positions 1103-1267) was found in two infantile patients, producing an abnormally large mRNA; one patient was probably a homozygote, and the other was a heterozygote of this mutation. The other two infantile patients had different mutations; a 123 Gly(GGG)----Arg(AGG) mutation in one patient and a 316 Tyr(TAT)----Cys(TGT) mutation in the other. A 201 Arg(CGC)----Cys(TGC) mutation, eliminating a BspMI site, was detected in a late-infantile/juvenile patient; the restriction-site analysis of amplified genomic DNA confirmed his heterozygosity for this mutation. A 51 Ile(ATC)----Thr(ACC) mutation was found in all five adult/chronic patients examined in this study. It created a SauI site, and restriction-site analysis confirmed that four patients were homozygous mutants. The other was a compound heterozygote for this mutation and another 457 Arg(CGA)----Gln(CAA) mutation. These mutant genes expressed markedly decreased or completely deficient enzyme activities in beta-galactosidase-deficient human fibroblasts transformed by adenovirus-SV40 recombinants. We conclude that gene mutations are heterogeneous in GM1-gangliosidosis but that the 51 Ile(ATC)----Thr(ACC) mutation is common among the Japanese adult/chronic cases. Genotype-phenotype correlations in GM1-gangliosidosis are briefly discussed.

Topics: Adolescent; Adult; Alleles; Base Sequence; beta-Galactosidase; Cells, Cultured; Child; Chronic Disease; Cloning, Molecular; G(M1) Ganglioside; Gangliosidoses; Genes; Humans; Infant; Japan; Molecular Sequence Data; Mutation; Oligonucleotide Probes; Polymerase Chain Reaction; RNA, Messenger

Sheep affected with ovine GM1 gangliosidosis are normal at birth and develop clinical signs, initially ataxia, commencing at approximately 5 months of age, which progresses rapidly to recumbency. Superovulation and embryo transfer techniques were applied to a flock of carrier sheep of ovine GM1 gangliosidosis to increase the numbers of carrier and affected animals. A recipient ewe with 3 at-risk fetuses died at 4 months of gestation (normal ovine gestation is 5 months), and spectrofluorimetric assay of cerebral lysosomal beta-galactosidase of the fetuses showed that 2 were carriers and one was an affected fetus. The affected fetus had marked cytoplasmic enlargement and vacuolization of central and peripheral nervous system neuronal soma and of hepatocytes and renal epithelial cells. Lectin histochemistry indicated abnormal storage of complex carbohydrates, with terminal saccharide moieties consisting of beta-galactose, N-acetylneuraminic acid, and N-acetylgalactosamine. This case underlines the need for prenatal initiation of therapy and also demonstrates that vacuolization alone is not the cause of clinical signs in this lysosomal storage disease in that clinical signs do not commence until at least 5 months after vacuolization is histologically apparent.

Topics: Acetylgalactosamine; Animals; beta-Galactosidase; Embryo Transfer; Female; Fetus; G(M1) Ganglioside; Galactose; Gangliosidoses; Heterozygote; Lysosomes; N-Acetylneuraminic Acid; Pregnancy; Prenatal Diagnosis; Sheep; Sheep Diseases; Sialic Acids

GM1-gangliosidosis is a genetic neurological disorder caused by mutations in the lysosomal acid beta-galactosidase gene. While its phenotypic expression is complex, it is usually classified as being of infantile, juvenile, or adult form, on the basis of age at onset, the rate of symptomatic progression, and severity of central nervous system involvement. We have analyzed the acid beta-galactosidase gene in 12 Japanese patients from nine families. The aim was to identify mutations in individual patients and then to examine possible correlation between the mutations and the clinical phenotypes. Northern blotting studies with a full-length human beta-galactosidase cDNA showed that the mRNA ranged from undetectable to substantially decreased in the infantile patients but was normal in quantity and size in all juvenile and adult patients. Four distinct missense mutations have been identified, each limited to the respective clinical forms within our small-size samples. In the infantile patient with decreased but detectable mRNA, a point mutation was found resulting in Arg49----Cys. In the infantile patient with nearly undetectable mRNA, mutation Arg457----Ter was identified. The mutation Arg201----Cys was found in all four of the juvenile patients, while all six adult patients were homozygous for the point mutation Ile51----Thr. The mutations found in the juvenile and adult patients alter restriction sites in the normal gene and thus are amendable to quick screening. The prediction that these mutations are responsible for the clinical disease was confirmed by no expression of the catalytic activity of the mutant proteins in the COS-I cell expression system.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Base Sequence; beta-Galactosidase; Blotting, Northern; Cell Line; Child, Preschool; Cloning, Molecular; Female; G(M1) Ganglioside; Gangliosidoses; Humans; Infant; Japan; Male; Molecular Sequence Data; Mutation; Polymerase Chain Reaction; Transfection

Asparagine-linked sugar chains of sphingolipid activator protein 1 (SAP-1) purified from normal human liver and GM1 gangliosidosis (type 1) liver were comparatively investigated. Oligosaccharides released from the two SAP-1 samples by hydrazinolysis were fractionated by paper electrophoresis and by Aleuria aurantia lectin-Sepharose and Bio-Gel P-4 (under 400 mesh) column chromatography. Structures of oligosaccharides in each fraction were estimated from data on their effective molecular sizes, behavior on immobilized lectin columns with different carbohydrate-binding specificities, results of sequential digestion by exoglycosidases with different aglycon specificities, and methylation analysis. Sugar chains of SAP-1 purified from normal human liver and from GM1 gangliosidosis (type 1) liver were different from each other, although both of them were derived from complex-type sugar chains. The sugar chains of the former were the following eight degradation products from complex-type sugar chains by exoglycosidases in lysosomes: Man alpha 1----6(Man alpha 1----3)Man beta 1----4GlcNAc beta 1----4GlcNAcOT, Man alpha 1----6(Man alpha 1----3)Man beta 1----4GlcNAc beta 1----4(Fuc alpha 1----6)GlcNAcOT, Man alpha 1----6Man beta 1----4GlcNAc beta 1----4GlcNAcOT, Man alpha 1----6Man beta 1----4GlcNAc beta 1----4(Fuc alpha 1----6)GlcNAcOT, Man beta 1----4GlcNAc beta 1----4GlcNAcOT, Man beta 1----4GlcNAc beta 1----4(Fuc alpha 1----6)GlcNAcOT, GlcNAc beta 1----4GlcNAcOT, and GlcNAcOT. In contrast to these, the sugar chains of the latter were sialylated and nonsialylated mono- to tetraantennary complex-type sugar chains that were not fully degraded due to a metabolic defect in acid beta-galactosidase activity.

Topics: Asparagine; Carbohydrate Conformation; Carbohydrate Sequence; Chromatography, Affinity; G(M1) Ganglioside; Gangliosidoses; Glycoproteins; Glycoside Hydrolases; Humans; Liver; Molecular Sequence Data; Neuraminidase; Oligosaccharides; Reference Values; Saposins; Sphingolipid Activator Proteins

A female infant with early-onset GM1-gangliosidosis type I was investigated. The lymphocytes, transformed lymphocytes and cultured skin fibroblasts of the patient were demonstrated to have severe beta-D-galactosidase deficiency. The beta-D-galactosidase activities of these cells from the patient's father and mother were at the lower limit of the normal range. The oligosaccharide accumulation in urine of the patient showed the typical type I GM1-gangliosidosis pattern, but no GM1 ganglioside was detected in the patient's urine or transformed lymphocytes. The clinical features were compatible with infantile GM1-gangliosidosis. The mixture of homogenates from the cultured fibroblasts or transformed lymphocytes of the patient and controls showed no complementation of beta-D-galactosidase activity against the controls.

Topics: beta-Galactosidase; Cells, Cultured; Chromatography, Thin Layer; Female; Fibroblasts; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Glycosphingolipids; Hexosaminidases; Humans; Infant; Lymphocyte Activation; Lymphocytes; Skin

Allogeneic bone marrow transplantation was carried out in an 81-day-old Portuguese water dog with GM1 gangliosidosis using a DLA identical sibling as donor. Engraftment was complete and beta-galactosidase activity in leukocytes of the transplanted dog were similar to those in the donor. Over the next 2.5 months neurological deterioration in the transplanted dog was similar to that in untreated dogs with GM1 gangliosidosis. Cerebral ganglioside GM1 concentrations were not diminished by bone marrow transplantation and cerebral beta-galactosidase activity was negligible. We conclude that allogeneic bone marrow transplantation early in life is ineffective in canine GM1 gangliosidosis.

Topics: Animals; beta-Galactosidase; Bone Marrow Transplantation; Brain; Chromatography, Thin Layer; Dogs; Female; G(M1) Ganglioside; Gangliosidoses; Genetic Carrier Screening; Neurologic Examination

Clinical and biochemical studies are reported on a 32-year-old man with GM1 gangliosidosis who presented with a slowly progressive dystonia that began when he was aged 7 years and eventually became almost totally incapacitating at the age of 35. There was only mild intellectual deterioration, but myoclonus, seizures and macular cherry-red spots were never observed. Proton-density and T2-weighted MRI scans showed symmetrical hyperintense lesions of both putamina. No increase of GM1 ganglioside was found in plasma or cerebrospinal fluid, and the metabolism of GM1 ganglioside in cultured skin fibroblasts from the patient was also almost normal, although the residual activity of GM1 ganglioside beta-galactosidase activity was only 10% of normal. These findings suggest that impaired GM1 ganglioside metabolism is not present systemically as it is in the infantile and juvenile types of the disorder, but is mainly confined to the central nervous system in chronic GM1 gangliosidosis.

Topics: Adult; Chronic Disease; Dystonia; G(M1) Ganglioside; Gangliosidoses; Humans; Male

Topics: Alkaline Phosphatase; Bone Marrow; Bone Marrow Cells; Bone Marrow Examination; Foam Cells; G(M1) Ganglioside; Gangliosidoses; Humans; Infant; Infant, Newborn; Male; Osteoblasts; Osteoclasts

Urinary oligosaccharides of GM1 gangliosidosis patients (type 2A, 4 cases; type 2B, 2 cases) were investigated using the Bio Gel system. The levels of urinary oligosaccharide excreted (nmol/mg creatinine) by the type 2A patients were 4.1 times the levels of the type 2B patients. Patients of type 2A excreted high molecular weight oligosaccharides which were not detected in the urine of type 2B, and excreted oligosaccharides with long linkages of repeating structures. Thus, type 2A apparently has biochemically different characteristics from type 2B related to urinary oligosaccharide. Differentiation of type 2A from type 2B can thus be made biochemically. The structures of 5 different kinds of oligosaccharides not reported previously were confirmed.

Topics: Adolescent; Adult; Carbohydrate Sequence; Child; Child, Preschool; G(M1) Ganglioside; Gangliosidoses; Gas Chromatography-Mass Spectrometry; Humans; Molecular Sequence Data; Oligosaccharides

In order to delineate clinical subtypes of GM1 gangliosidosis enzymologically, we prepared galactosyl oligosaccharides from the urine of patients, as substrates, and established the method of the galactosyl oligosaccharide beta-galactosidase assay. Galactosyl oligosaccharides beta-galactosidase activities (nmol/mg protein/20 h) in vitro, using substrates without repeating structures were; type 1, 1.0 +/- 0.5 (n = 6), type 2A, 2.1, type 2B, 3.4 +/- 0.7 (n = 5), type 3, 4.9 +/- 0.2 (n = 2). The activities in vitro using substrates with repeating structures were: type 1, 0.3 +/- 0.2 (n = 5), type 2A, 1.2, type 2B, 2.2 +/- 0.5 (n = 4), type 3, 4.2 +/- 0.3 (n = 2). The activities using substrates with and without repeating structures were affected in the fibroblasts of patients, and the residual activities in each subtype were reduced progressively with the increasing severity of the clinical features. The ratio between activities using substrates without repeating structures and activities using substrates with repeating structures indicated that beta-galactosidase activities toward Gal beta 1- of repeating structures were reduced progressively with the increasing severity of the clinical features. The activities in vivo (pmol/mg protein per 24 h) were: type 1, 11.8 +/- 1.8 (n = 2), type 2A, 24.8, type 2B, 40.0 +/- 9.7 (n = 2), type 3, 63.2. The activities in vivo were affected in the fibroblasts of patients and the residual activities were reduced in proportion to the severity of the clinical features. These differences of residual activities among each subtype make it possible to delineate clinical subtypes enzymologically.

Topics: beta-Galactosidase; Child; Fibroblasts; G(M1) Ganglioside; Gangliosidoses; Humans; Oligosaccharides; Skin

We report a case of a 10-month-old male infant with GM1 type 1 gangliosidosis who also had hyperpigmented macules and patches. Light and electron microscopic findings correlated with previously published reports on findings in skin biopsy specimens of patients with lipid storage disorders. The hyperpigmented macules are most likely mongolian spots. A differential diagnosis of these lesions is discussed.

Topics: G(M1) Ganglioside; Gangliosidoses; Humans; Infant; Male; Microscopy, Electron; Pigmentation Disorders; Skin

A six-month-old female gypsy child, the daughter of second degree cousins, born after a full-term pregnancy and normal delivery, is described. There was generalized neonatal edema. Abnormalities included psychomotor retardation from birth and progressive appearance of facial dysmorphism, organ enlargement, axial hypotonia, hypertonia in limbs, myoclonic jerks, optic atrophy and bilateral cherry-red spots. The diagnosis of GM1 type 1 gangliosidosis was confirmed by biochemical, enzymatic and ultrastructural findings.

Topics: Biopsy; Bone Diseases; Edema; Female; G(M1) Ganglioside; Gangliosidoses; Humans; Infant; Maxillofacial Development; Psychomotor Disorders; Skin

A patient with severe deficiency of beta-galactosidase, who developed skin lesions of angiokeratoma corporis diffusum between the 3rd and 10th month of life, is described. The activity of other lysosomal enzymes, including alpha-neuraminidase, was normal. The first signs of the disease were noticed during the first month of life. By 3 months coarseness of the face and psychomotor retardation were present. In addition to angiokeratoma, he had large mongolian spots and several scattered slate-blue spots of pigmentation over his body. With the exception of the skin lesions, the other clinical signs and the course of the psychomotor deterioration were within the clinical picture of GM1 gangliosidosis, Type 1. Angiokeratoma, a manifestation of several lysosomal disorders, may appear in GM1 gangliosidosis during the first year of life.

Topics: beta-Galactosidase; Fabry Disease; G(M1) Ganglioside; Gangliosidoses; Humans; Infant; Male; Skin

A case of beta galactosidase deficiency is described in a 20-month-old boy. The child was hospitalized at 4 months of age for malabsorption syndrome. Biopsies of the small intestine and liver were performed and electron microscopy of the liver specimens strongly suggested a gangliosidosis. The cytoplasm of macrophages, Kupffer cells and hepatocytes contained membrane-bound lysosomes with a granular, fibrillar appearance and tubular structures interpreted as ganglioside deposits. Enzymatic deficiency was confirmed by biochemical investigation of leukocytes from both the patient and members of his immediate family. Although visceromegaly is typical of Landing disease, symptoms of malabsorption and hypertension have not been reported in its course.

Topics: Edema; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Humans; Hypoproteinemia; Infant; Intestine, Small; Liver; Malabsorption Syndromes; Male; Microscopy, Electron; Mucous Membrane; Radiography

alpha-Galactosidase and beta-galactosidase activities have been determined in leukocyte preparations from 100 randomly selected Chinese adults. For alpha-galactosidase, two groups with low activities were identified: group I consisted of 3 females having activities below 40% of normal, and group II consisted of 5 males and 1 female with activities about 60% of normal. Family studies suggested that these low alpha-galactosidase activities are genetically determined. Only 1 individual was found to have about 50% of normal beta-galactosidase activity; presumably he is a carrier for beta-galactosidase deficiency (GM1 gangliosidosis).

Topics: Adult; alpha-Galactosidase; beta-Galactosidase; China; Fabry Disease; Female; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Genetic Carrier Screening; Humans; Leukocytes; Lysosomes; Male; Pedigree; United States

beta-Galactosidases were purified to homogeneity from livers of a normal control and a patient with the adult form of GM1 gangliosidosis. The purification was achieved by chromatography on DEAE-Sepharose fast flow, Con A-Sepharose, p-aminophenyl-1-thio-beta-D-galactopyranoside-Sepharose, and QAE-Mono Q. The normal and mutant enzymes were purified about 5000-fold with a yield of 10% and 1800-fold with a yield of 34%, respectively, and could hydrolyze 4-methylumbelliferyl-beta-D-galactoside, GM1 ganglioside, and asialofetuin. The purified normal enzyme was eluted from a TSK gel G-4000SW column as three symmetrical peaks of protein which were coincident with the three peaks of enzyme activity. The enzyme in these three peaks had apparent molecular weights of 800,000 (polymer), 140,000 (dimer), and 65,000 (monomer), whereas the mutant enzyme was eluted as two symmetrical peaks of protein and enzyme activity. The apparent molecular weight of a major monomeric form of the enzyme (beta-galactosidase A) was 60,000, and no dimeric form of the enzyme existed. Normal and mutant purified enzyme preparations migrated as a single major protein band with apparent molecular weights of 65,000 or 60,000, respectively, by SDS-polyacrylamide gel electrophoresis after treatment with mercaptoethanol. On isoelectric focussing, the mutant enzyme migrated more anodally than the normal enzyme. The mutant enzyme also had altered enzyme properties, such as pH optimum, Km values, substrate specificity and heat-stability. These data on the characteristics of the purified enzyme preparations provide the first direct evidence that patients with the adult form of GM1 gangliosidosis have a structurally altered beta-galactosidase.

Topics: Adult; beta-Galactosidase; Enzyme Stability; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Humans; Kinetics; Liver; Mutation; Reference Values; Substrate Specificity

GM1 gangliosidosis in the infantile form is a rapidly fatal storage disease produced by deficiency of acid beta-galactosidase. Ultrastructural studies of the eyes from two fetuses affected with GM1 gangliosidosis were performed in an effort to assess tissue-specific distribution of storage inclusions in the different ocular components derived from neuroectoderm, surface ectoderm, and mesoderm. Two major configurations of inclusions were observed: electronlucent vacuoles and pleiomorphic osmiophilic membranes. Although the latter changes mainly affected the retinal neurons, they were occasionally found in cells of epithelial and mesenchymal origin. The findings indicate that the lysosomal storage process in GM1 gangliosidosis, type 1, has a wide morphologic spectrum that is already present in the early period of fetal life.

Topics: Diagnosis, Differential; Eye; Female; Fetal Diseases; G(M1) Ganglioside; Gangliosidoses; Humans; Microscopy, Electron; Pregnancy; Pregnancy Trimester, Second; Tay-Sachs Disease

Three sisters (ages 27, 24, and 17 years) presented with slowly progressing dystonic dementia and spastic tetraparesis with infantile onset. CSF, bone marrow, and conjunctival cells showed storage vacuoles. Biochemical analysis revealed increased urinary oligosaccharide excretion and decreased activity of acid beta-D-galactosidase and beta-D-fucosidase in serum, leukocytes, and cultured fibroblasts. The parents' enzyme values were in the heterozygous range. This is the only case in the literature of severe dementia associated with the clinical symptoms of type 3 GM1 gangliosidosis. The clinical heterogeneity of GM1 gangliosidosis and the significance of the combination of beta-D-galactosidase and beta-D-fucosidase defects in this syndrome are discussed.

Topics: Adolescent; Adult; alpha-L-Fucosidase; beta-Galactosidase; Bone Marrow; Cells, Cultured; Conjunctiva; Dementia; Female; Fibroblasts; G(M1) Ganglioside; Gangliosidoses; Humans; Leukocytes; Lysosomes; Microscopy, Electron; Oligosaccharides; Vacuoles

Three Portuguese water dog siblings, all females aged 5 to 7 months, were killed following a brief period of neurologic disease. Tissues were processed for light and electron microscopy and for biochemical analyses. All pups had membranous cytoplasmic inclusions in neurons throughout the brain and spinal cord. Cytoplasmic vacuoles were present in cells of many organs outside the nervous system. GM1 ganglioside in brain was markedly elevated in all three dogs, and beta-galactosidase activity was less than 10% of control values. These findings are similar to those in GM1 gangliosidosis of man and animals although the number of organs and tissues containing vacuolated cells is greater.

Topics: Animals; Brain; Dog Diseases; Dogs; Female; G(M1) Ganglioside; Gangliosidoses

Lectin histochemical studies were performed on paraffin-embedded brain, spinal cord and eye sections of 16 animals from four different species affected with GM1- and GM2-gangliosidosis to identify specific carbohydrate residues in the perikaryon of neurons. We examined tissues from cats, cattle and dogs with GM1-gangliosidosis and from cats, dogs, and swine with GM2-gangliosidosis and compared them to corresponding normal animals. In all but two cases, the neurons stained intensely with Concanavalia ensiformis agglutinin (Con A); in 12 cases they stained with Dolichos biflorus agglutinin; in 10 cases with Ulex europaeus agglutinin-I; in 9 cases with Griffonia simplicifolia-I; and 8 with soybean agglutinin. Neurons from control tissues stained weakly with Con A, but not with any of the other lectins used. Similar staining patterns of neurons were noted in animals affected with the same disorder originating from the same mutant line. These findings highlight the fact that in gangliosidosis, the lectin staining patterns of neurons may be influenced by the deficiency in enzyme activity and by additional unknown but inherited factors.

Topics: Animals; Brain; Cats; Cattle; Concanavalin A; Dogs; Endothelium; G(M1) Ganglioside; G(M2) Ganglioside; Gangliosides; Gangliosidoses; Lectins; Plant Lectins; Species Specificity; Swine; Wheat Germ Agglutinins

Topics: Adult; Animals; beta-Galactosidase; Child; Child, Preschool; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Humans; Lysosomes

This study was designed to establish an in vitro model with biochemical and morphological similarities to the human neurodegenerative disease GM1 gangliosidosis. Utilizing a specific inactivator of the lysosomal enzyme GM1-ganglioside beta-galactosidase (beta-D-galactopyranosylmethyl-p-nitrophenyltriazene [beta-GalMNT]) and neuroblastoma X glioma hybrid cells (NG108-15), we suppressed beta-galactosidase activity for up to 72 hours. Coincidental with suppression of this enzyme to levels less than 1% of control, we found up to a nine-fold accumulation of its substrate, the GM1-ganglioside, and the ultrastructural appearance of membranous cytoplasmic bodies. beta-GalMNT treatment suppressed growth but had little effect on the specific activity of choline acetyltransferase, lactate dehydrogenase, or other lysosomal enzymes including galactosylceramidase. This model should permit studies of the neurophysiological effects of increased ganglioside accumulation and their reversibility.

Topics: beta-Galactosidase; Brain; Brain Diseases, Metabolic; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Humans; Inclusion Bodies; Lysosomes; Triazenes

Confluent cultures of human skin fibroblasts were maintained for 10 days with sphingosine labeled [3H]GM2. Labeled medium was then replaced with normal medium and the cells maintained for 42 days with weekly medium changes. Cells were harvested at regular intervals and cells, medium, and trypsin digest supernatant analyzed for [3H]GM2 and its metabolic products. The ganglioside can be membrane associated and removed by trypsin, or membrane incorporated and trypsin insensitive. The membrane incorporated material is apparently transported to the lysosomes slowly by membrane flow, where 80% of the cellular GM2 can be metabolized by day 42. [3H]GM2 as well as its metabolic products in control cells is continuously released into the medium, during which it can also become associated with the cell surface membrane. There is no detectable metabolism of the [3H]GM2 in GM2 gangliosidosis cell lines over the extended post-labeling period, indicating that there is no residual enzyme activity in these cells. Undegraded GM2 is continuously released into the medium and remains associated with the cell surface membrane as well.

Topics: Fibroblasts; G(M1) Ganglioside; Gangliosidoses; Glycine; Humans; Kinetics; Protein Biosynthesis; Reference Values; Skin; Tritium

The observation of generalized GM1 gangliosidosis type 1 (Norman-Landing disease) is reported. The case is typical, featuring all the main clinical and biological signs of the disease. Diagnosis was established by the demonstration of a severe deficit in beta-galactosidase activity in leucocytes, by the demonstration of oligosaccharides in the urine, and by the histological examination after the fatal outcome before the age of two with severe respiratory distress.

Topics: Cerebral Cortex; G(M1) Ganglioside; Gangliosidoses; Humans; Infant; Male; Nerve Degeneration; Oligosaccharides

Systematic Golgi studies have been performed on major subcortical, diencephalic, brain stem and spinal cord regions from cats with the inherited neuronal storage disease, GM1 gangliosidosis. Resulting data have been compared with other Golgi studies of neuronal storage disorders in man and animals, including an earlier, more limited examination of this same disease model. These previous studies have shown that in human and feline gangliosidoses cortical pyramidal neurons undergo remarkable changes in soma-dendritic geometry. The latter include the formation of conspicuous cellular enlargements between somata and axonal initial segments (meganeurites) and the sprouting of secondary neuritic processes from this same region of the cell. Further, ultrastructural studies have revealed normal appearing synapses on the surface of this ectopically placed dendritic-like membrane. Results of the present study indicate that the distribution of meganeurites, secondary neurites and other geometrical distortions of neurons in GM1 gangliosidosis varies with cell type and brain region. This cell type-specific response to the metabolic error and subsequent storage could be categorized in three ways. Firstly, certain types of cells (e.g. multipolar neurons of the amygdala and claustrum) exhibited changes similar to those reported for cortical pyramidal neurons. That is, cells of these regions either displayed spine or neurite-bearing meganeurites, or enlarged axon hillocks which were covered with similar processes. Other types of neurons did not demonstrate ectopic neurite growth or spine-covered meganeurites, but did display prominent aspiny meganeurites (e.g. neurons of the superior colliculus, periaqueductal gray, hypothalamus and basal forebrain nuclei). A third category of neurons did not possess meganeurites or neurite growth but instead demonstrated massive somatic expansion which exceeded that observed in meganeurite-bearing cell types (e.g. certain brain stem and spinal cord neurons). These data have been compared with the more limited Golgi studies of other types of neuronal storage disorders and the same types of neurons appeared to respond in similar fashion across this spectrum of diseases. The data presented and discussed in this paper demonstrate three significant morphological events which occur in neurons as a result of lysosomal hydrolase deficiency. These are storage, which occurs in all neurons but manifests as meganeurite formation or somatic enlargeme

Topics: Animals; Cats; Central Nervous System; Dendrites; G(M1) Ganglioside; Gangliosidoses

By using cholera toxin B subunit and its antibody, the deposition of GM1-ganglioside in the cerebral cortex and peripheral nerves including Meissner and Auerbach's plexuses in the intestine and other visceral nerves of generalized GM1-gangliosidosis was demonstrated. The GM1-ganglioside was found in the swollen neurons of cerebral cortex and ganglion cells of the peripheral nerves. Electron microscopically, parts of membranous cytoplasmic bodies, and amorphous substances among them, revealed a positive reaction for the cholera toxin staining.

Topics: Cerebral Cortex; Cholera Toxin; Female; G(M1) Ganglioside; Gangliosidoses; Humans; Microscopy, Electron; Myenteric Plexus; Pancreas; Peptide Fragments; Peripheral Nerves; Submucous Plexus

Ca2+ transport was studied in synaptosomes prepared from normal cats and cats with GM1 gangliosidosis. The influx of Ca2+ was found to be a biphasic process in synaptosomes from both GM1 mutant and normal cats. Both the fast and slow phases of voltage-dependent Ca2+ uptake were significantly reduced in cats with the lysosomal storage disease, however the inhibitory mechanisms differed. The fast phase of Ca2+ uptake was inhibited uncompetitively, whereas the slow phase was inhibited competitively. In addition, Na+-dependent Ca2+ efflux was reduced significantly in cats with GM1 gangliosidosis. Since it is well established that maintenance of Ca2+ homeostasis is essential for normal neuronal function, a ganglioside-induced disruption of Ca2+ transport across synaptic membranes may be responsible, in part, for the neuronal dysfunction characteristic of GM1 gangliosidosis.

Topics: Animals; Calcium; Calcium Radioisotopes; Cats; Choline O-Acetyltransferase; G(M1) Ganglioside; Gangliosidoses; Kinetics; Motor Cortex; Mutation; Synaptosomes

We report a case of GMI gangliosidosis type I with cherry-red spot, optic atrophy, and corneal cloudings. The diagnosis was confirmed by the deficiency of beta-galactosidase enzyme in leucocytes and in urine.

Topics: beta-Galactosidase; Cornea; Corneal Opacity; G(M1) Ganglioside; Gangliosidoses; Humans; Infant; Macula Lutea; Male; Optic Atrophy

Topics: Adaptation, Psychological; Family; Family Therapy; G(M1) Ganglioside; Gangliosidoses; Genetic Counseling; Humans; Pedigree; Risk Factors

Ferric ion-ferrocyanide staining and safranin-0-counterstaining of neocortical tissue from cats with GM1 gangliosidosis have established that pyramidal neuron meganeurites occur proximal to axonal initial segments and that they are distinct from axonal spheroids. The latter, which were found to be widely distributed throughout cerebral cortex, were located distal to axonal initial segments and could be differentiated from meganeurites at both light and electron microscopic levels. This report confirms an earlier electron microscopic study which suggested that meganeurites are of axon hillock origin, and illustrates the striking distinction between abnormalities in the soma-dendritic and axonal domains of neurons in a lysosomal storage disease.

Topics: Animals; Axons; Cats; Cerebral Cortex; Chlorides; Dendrites; Ferric Compounds; Ferrocyanides; G(M1) Ganglioside; Gangliosidoses; Microscopy, Electron; Neurons; Phenazines; Staining and Labeling

The uptake and degradation of GM1 ganglioside (GM1) and asialoGM1 ganglioside (GA1) were studied in cultured fibroblasts from normal individuals and patients with beta-galactosidase deficiency, using the lipid-loading test. The glycolipids were incorporated from the media into the fibroblasts and the terminal galactose was hydrolyzed in normal cells. The hydrolysis rates of GA1 were 80-86% of normal on the 3rd day after loading, while GM1 was hydrolyzed slowly; 35-54% on the 14th day. In infantile GM1 gangliosidosis and I-cell disease, little GM1 and GA1 was hydrolyzed on any day of culture, while fibroblasts from patients with adult GM1 gangliosidosis, Morquio disease type B and galactosialidosis hydrolyzed the lipids at nearly normal rates. The intracellular accumulation of the glycolipids, on the basis of protein content, was abnormally high in the case of infantile GM1 gangliosidosis and I-cell disease, but normal in the other disorders examined. These observations indicate that the in situ metabolism of GM1 and GA1 is probably normal in fibroblasts from patients with adult GM1 gangliosidosis, Morquio disease type B and galactosialidosis, although in vitro beta-galactosidase activities in these disorders are very low. The results are compatible with findings that GM1 and GA1 do not accumulate in the somatic organs of patients with adult GM1 gangliosidosis and galactosialidosis. In I-cell disease, however, the results of the loading test did not agree with the finding that there is little accumulation of glycolipids in postmortem tissues.

Topics: Adult; Animals; beta-Galactosidase; Brain; Cattle; Cells, Cultured; Fibroblasts; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Glycosphingolipids; Humans; Infant; Kinetics; Reference Values; Tritium

Sphingolipid activator proteins (SAP) are relatively low-molecular-mass proteins that stimulate the hydrolysis of specific sphingolipids by the required lysosomal enzymes. SAP-1 or sulfatide/GM1 ganglioside activator protein has previously been demonstrated to stimulate the enzymatic hydrolysis of sulfatide, GM1 ganglioside and globotriaosylceramide. Using monospecific rabbit antibodies against human liver sulfatide/GM1 activator, the biosynthesis and processing of this activator were studied in cultured skin fibroblasts from controls and patients with GM1 gangliosidosis and a variant form of metachromatic leukodystrophy. When [35S]methionine was presented in the medium to control human fibroblasts for 4 h, the majority of the immunoprecipitable radiolabeling was confined to bands within three regions of apparent molecular mass 65-70, 35-52 and 8-13 kDa. The only immunoprecipitable radiolabeled species excreted into the medium when NH4Cl was present had an apparent molecular mass of 70 kDa. When the excretion products were given to fresh cells followed by incubation for up to 24 h there was production of the mature species. Treatment of the 70 kDa form with endoglycosidase F resulted in production of a 53 kDa molecular mass form. Pulse-chase experiments indicated that the initial immunoprecipitable translation product was 65 kDa which increased to 70 kDa over the next hour. The 65 kDa species must result from co-translational glycosylation of the polypeptide chain. Apparently, intralysosomal processing converts the 13 kDa form to the 8-11 kDa species. The cells from the patient with GM1 gangliosidosis could not process to the smallest species found in controls due to the deficiency of acid beta-galactosidase. Patients who have a variant form of metachromatic leukodystrophy do not make any immunoprecipitable radiolabeled products in the cells or in the media. This indicates a severe mutation in the gene coding for this activator protein. The production of such small mature species from a relatively large precursor form may regulate the production of this interesting protein.

Topics: Cells, Cultured; Electrophoresis, Polyacrylamide Gel; Endocytosis; Fibroblasts; G(M1) Ganglioside; Gangliosidoses; Glycoproteins; Glycoside Hydrolases; Humans; Immunochemistry; Leukodystrophy, Metachromatic; Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; Methionine; Mucolipidoses; Mutation; Photofluorography; Protein Biosynthesis; Proteins; Saposins; Sphingolipid Activator Proteins

Cholinergic processes were measured in motor cortex, hippocampus, and striatum of cats in the terminal stages of GM1 gangliosidosis and compared to those of control cats. The greatest difference observed was elevation in the rate of K+-stimulated release of acetylcholine (ACh) from brain slices prepared from affected cats. The K+-stimulated release of endogenous ACh was increased by 31-43% and of newly synthesized ACh by 19-80% in brain slices from different brain regions. All regions that were examined were affected but the greatest effects occurred in cortex. The rate of synthesis of ACh was elevated in cortical and hippocampal slices. Choline acetyltransferase activity in brain regions of cats with GM1 gangliosidosis was not significantly different from that in controls, whereas high-affinity choline transport in cortical synaptosomes was elevated. Muscarinic receptor binding sites were reduced in the cortex, hippocampus, and striatum of GM1 mutant cats, whereas the apparent affinity was not altered. These results indicate that there are major alterations of cholinergic function in the brains of cats with GM1 gangliosidosis.

Topics: Acetylcholine; Animals; Biological Transport; Brain; Cats; Choline; Choline O-Acetyltransferase; Corpus Striatum; Disease Models, Animal; G(M1) Ganglioside; Gangliosidoses; Hippocampus; Motor Cortex; Potassium; Receptors, Muscarinic

Immunoelectron microscopy was performed to study the biosynthesis of lysosomal beta-galactosidase (beta-gal) in normal and mutant human fibroblasts. Using polyclonal and monoclonal antibodies we show in normal cells precursor forms of beta-gal in the rough endoplasmic reticulum (RER) and in the Golgi apparatus throughout the stack of cisternae. In the lysosomes virtually all beta-gal exists as a high molecular weight multimer of mature enzyme. In the autosomal recessive disease GM1-gangliosidosis caused by a beta-gal deficiency and in galactosialidosis, associated with a combined deficiency of lysosomal neuraminidase and beta-gal, precursor forms of the latter enzyme are found in RER, Golgi and some labeling is present at the cell surface. The lysosomes remain unlabeled, indicative for the absence of enzyme molecules in this organelle. In galactosialidosis fibroblasts also no mature beta-gal is found in the lysosomes but in these cells the presence of the monomeric form can be increased by leupeptin (inhibition of proteolysis) whereas addition of a partly purified 32 kDa "protective protein" results in the restoration of high molecular weight beta-gal multimers in the lysosomes.

Topics: Antibodies; Antibodies, Monoclonal; beta-Galactosidase; Fibroblasts; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Histocytochemistry; Humans; Immunochemistry; Leupeptins; Lysosomes; Metabolism, Inborn Errors; Microscopy, Electron; Neuraminidase; Subcellular Fractions

A highly sensitive microassay method and a microscale purification system were developed to isolate the residual acid beta-galactosidase in GM1-gangliosidosis fibroblasts. The sensitivity of the microassay system, composed of a 96-well microplate and a microplate fluorometer, was 100-fold higher than that of the conventional system and the response was linear in the pmole range. Acid beta-galactosidase was characterized as a thiol enzyme which was inactivated by a mercuric compound. This enzyme was completely adsorbed on an Hg-agarose column and was easily eluted from the column by 10 mM 2-mercaptoethanol. The microscale purification system using Con A-Sepharose, PAT-Sepharose, and Hg-agarose column chromatography achieved 565- and 7,970-fold purifications of acid beta-galactosidase with an overall yields of 44% and 45% from normal and GM1-gangliosidosis fibroblasts, respectively. The purified enzyme fractions did not contain any other lysosomal enzyme activities except for a small amount of beta-N-acetylhexosaminidase activity.

Topics: beta-Galactosidase; Chromatography, Agarose; Fibroblasts; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Humans; Mercury; Microchemistry

Cultured fibroblasts from different variants of GM1-gangliosidosis synthesize normal amounts of 88-kDa beta-galactosidase precursor. Yet the amount of the mature 64-kDa form is reduced to 5-15% of normal values. In this communication it is shown that the mutation in the infantile and adult form of GM1-gangliosidosis interferes with the phosphorylation of precursor beta-galactosidase. As a result the precursor is secreted instead of being compartmentalized into the lysosomes and further processed. The impaired phosphorylation might be due to conformational changes of the precursor molecule.

Topics: beta-Galactosidase; Cells, Cultured; Enzyme Precursors; Fibroblasts; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Humans; Molecular Weight; Phosphates; Phosphorus Radioisotopes; Phosphorylation; Protein Processing, Post-Translational; Skin

The uptake and catabolism of [3H-ceramide]-GM1 was followed in living fibroblasts from patient with different forms of beta-galactosidase deficiency. Gangliosides are identified according to the nomenclature of Svennerholm (1963). A total inability to metabolize the ingested substrate was found in infantile GM1-gangliosidosis whereas cells from an adult GM1-gangliosidosis variant showed a slower rate of degradation, compared with controls. Morquio B fibroblasts had a comparable catabolism of GM1 as controls. Fibroblasts from different types of galactosialidosis, a recessive disease associated with a coexistent beta-galactosidase/neuraminidase deficiency all showed degradation of ingested GM1. In view of the molecular defect in this disease, this catabolism must be due to the 10-20% of monomeric beta-galactosidase molecules present in the lysosomes. Unexpectedly, in these cells an impaired metabolism of GM3 was found. The same finding was observed when cells with an isolated neuraminidase deficiency (mucolipidosis I) were loaded with GM1. A hypothesis is presented to explain these results.

Topics: Adult; beta-Galactosidase; Cells, Cultured; Fibroblasts; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Humans; Infant; Mucolipidoses; Mucopolysaccharidosis IV

Cerebral lipids of patients with GM1-gangliosidoses, infantile, juvenile, and chronic type which are caused by deficiency of beta-galactosidase, were examined and compared to each other. The infantile type demonstrated abnormal accumulation of GM1 and asialo-GM1 in contrast with marked decrease in such major cerebral lipids as cholesterol, phospholipids, cerebroside, and sulfatide. It was also noted that significant amounts of such unusual lipids as free fatty acids, GlcCer, LacCer, GbOse3Cer, and GbOse-4Cer plus nLcOse4Cer were found in the brain. These findings pointed out that this infantile type might accompany a severe cerebral dysgenesis with poor myelination. The juvenile type also showed marked increase in GM1 and asialo-GM1, but the decrease in cholesterol, phospholipids, cerebroside, and sulfatide was not so much as the infantile type. These findings along with the occurrence of cholesterol ester suggested that the brain caused progressive demyelination after the immature myelin appeared. An autopsized brain tissue of a male patient who was eventually diagnosed as a case of GM1-gangliosidosis chronic type after his death, showed some accumulation of GM1 and asialo-GM1 particularly in the caudate nucleus and putamen, whereas it showed moderate amounts of GM1 in apparently normal gray and white matters. It seemed that there are no abnormal cerebral lipids except for gangliosides and some neutral glycosphingolipids in the chronic type.

Topics: Adult; Age Factors; Brain; Child, Preschool; Chronic Disease; Female; G(M1) Ganglioside; Gangliosidoses; Glycosphingolipids; Humans; Infant; Lipid Metabolism; Male; Phospholipids

A 24-week fetus with GM1 gangliosidosis (type 1) was studied using biochemical and histopathologic methods. Foam cells in viscera and placenta demonstrated widespread accumulation of a lipidlike material. By microscopy, central nervous system storage appeared confined to the retina and dorsal root ganglia, but the brain ganglioside content was measurably elevated compared with that of age-matched controls. These data, along with those of others, imply that, if the observed pathologic findings are irreversible, any attempts at intrauterine therapy must commence prior to the middle of the second trimester.

Topics: Abortion, Induced; Adult; Amniocentesis; beta-Galactosidase; Brain; Female; Fetal Diseases; G(M1) Ganglioside; Gangliosidoses; Humans; Kidney; Pregnancy

Among lipids, gangliosides can be selectively stained with Schiff's reagent if the oxidizing agent (sodium metaperiodate) is sufficiently dilute to exclude all but the readily oxidized sialic acid sugars. A borohydride-periodate-Schiff (BhPS) sequence is recommended as a reliable method, convenient to perform, for the detection of the intraneuronal lipid accumulations in the ganglioside storage disorders.

Topics: Borohydrides; Brain Chemistry; G(M1) Ganglioside; Gangliosides; Gangliosidoses; Histocytochemistry; Humans; Oxidation-Reduction; Periodic Acid-Schiff Reaction; Spinal Cord; Tay-Sachs Disease

Topics: beta-Galactosidase; Child, Preschool; G(M1) Ganglioside; Gangliosidoses; Humans; Male

To study the enzymatic properties of beta-galactosidase from the patients with a beta-galactosidase deficiency such as GM1 gangliosidosis, determination of enzymatic activity with naturally occurring substrates, asialofetuin in addition to another natural substrate, GM1 ganglioside, is essentially required. With a previously reported, simple and sensitive fluorometric assay for GM1 ganglioside beta-galactosidase using high performance liquid chromatography (HPLC), optimal reaction conditions were determined for the assay of acid beta-galactosidase activity toward asialofetuin in skin fibroblast homogenates. Under these conditions, reduced enzymatic activities could be detected in cultured skin fibroblasts from patients with type 1 and 3 GM1 gangliosidoses and mucopolysaccharidosis IV-B (Morquio B syndrome). This method was applicable to study of the enzymatic properties of the mutant beta-galactosidase and provided an alternative to assays employing radioactive or artificial substrates.

Topics: alpha-Fetoproteins; Asialoglycoproteins; beta-Galactosidase; Chromatography, High Pressure Liquid; Fetuins; Fibroblasts; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Humans; Hydrogen-Ion Concentration; Kinetics; Microchemistry; Skin; Spectrometry, Fluorescence

Acid hydrolases were isolated from the lysosome fraction of beta-galactosidase-deficient human fibroblasts and from the mannose 6-phosphate containing medium in which they were grown. Nearly half of the total beta-hexosaminidase and beta-glucuronidase from both sources bound to Ricin specifically. Lysosomal beta-hexosaminidase, metabolically labelled with [35S]-methionine, was also fractionated on Ricin-agarose. SDS-PAGE of immunoprecipitates from Ricin-binding and non-binding fractions revealed approximately equivalent amounts of cross-reacting material at the appropriate MW. We interpret these results to mean that acid hydrolases which are segregated to lysosomes are exposed to trans-Golgi processing enzymes to about the same extent as enzymes which are secreted, and that segregation by the Man 6-P receptor occurs after transit through the trans-Golgi compartment.

Topics: beta-N-Acetylhexosaminidases; Binding Sites; Cells, Cultured; Centrifugation, Density Gradient; Fibroblasts; G(M1) Ganglioside; Gangliosidoses; Glucuronidase; Golgi Apparatus; Hexosaminidases; Humans; Hydrolases; Immunochemistry; Lysosomes; Protein Binding; Protein Processing, Post-Translational; Ricin; Transferases

Neurochemical studies were performed on synaptosomal membranes from cats with GM1 or GM2 gangliosidosis to examine possible mechanisms of neuronal dysfunction in these disorders. The basic hypothesis tested was that deficient ganglioside catabolism causes increased ganglioside content of synaptosomal plasma membrane which in turn disrupts normal function. Fluidity characteristics of synaptosomal membranes were examined using fluorescence polarization. Results showed markedly reduced membrane fluidity in both GM1 and GM2 gangliosidosis. These results were supported by a second study which revealed that isolated synaptosomal membranes of GM1 gangliosidosis cats had a 24-fold increase in total ganglioside content caused predominantly by excess GM1, a 2.3-fold increased cholesterol content, and a 1.4-fold increased phospholipid content. Finally, kinetic analysis of synaptosomal plasma membrane Na+,K+-ATPase from cats with GM1 gangliosidosis showed negligible differences in kinetic parameters compared with controls. Thus, the enzyme appeared protected from the global membrane changes in fluidity and composition. These observations provide evidence for a pathogenetic mechanism of neuronal dysfunction in the gangliosidoses while demonstrating protection of certain vital functional components, such as Na+,K+-ATPase.

Topics: Animals; Brain; Cats; Cholesterol; Fluorescence Polarization; G(M1) Ganglioside; G(M2) Ganglioside; Gangliosidoses; Humans; Lipids; Membrane Fluidity; Phospholipids; Sodium-Potassium-Exchanging ATPase; Synaptic Membranes; Temperature

Postsynaptic potentials evoked by ventrolateral thalamic stimulation were recorded intracellularly from neurons in the precruciate cortex of GM1 mutants with HRP- or LY-loaded microelectrodes. Ganglioside-laden pyramidal neurons exhibiting somal distention and/or meganeurite formation were found to respond to thalamic stimulation with short duration IPSPs. Evoked EPSPs were recorded from two morphologically characterized large basket intrinsic neurons which deployed extensive intracortical axonal arborizations. These findings point to the preservation of intracortical inhibitory networks in the feline model of GM1 gangliosidosis, and to the possibility of abnormal integration of somadendritic inputs in ganglioside-laden pyramidal neurons.

Topics: Animals; Cats; Evoked Potentials; G(M1) Ganglioside; Gangliosidoses; Humans; Motor Cortex; Neural Inhibition; Synapses; Synaptic Transmission; Thalamic Nuclei

Topics: beta-Galactosidase; Cardiomyopathies; Female; Follow-Up Studies; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Heart Failure; Humans; Infant; Infant, Newborn; Male; Mitral Valve Insufficiency; Neuraminidase

Brains of two patients with GM1 gangliosidosis type 1 and type 2, together with the age-matched control brains, were analyzed for glycosphingolipids. Six species of neutral glycolipids, eight species of gangliosides, and sulfatide were isolated from the diseased brains and identified. In addition to GM1 ganglioside and its asialo derivative, the diseased brains accumulated considerable amounts of gangliotriaosylceramide and glycolipids belonging to the globo series, the accumulation of which cannot be explained by deficient beta-galactosidase activity in this disease. GM4 ganglioside was detected in the type 2 brain, but not in type 1. As to fatty acid composition of monohexosylceramides and sulfatide in the two diseased brains, stearic acid was more predominant in the type 1 brain than in the type 2 brain. In light of our previous observations on a Tay-Sachs brain and present results, it appears that metabolism of the globo series glycolipids, which is active in normal brain at early infancy but inactive thereafter, remains in brains with GM1 gangliosidosis (types 1 and 2) and Tay-Sachs disease, reflecting a disturbance in development of the brain.

Topics: beta-Galactosidase; Brain; Carbohydrates; Child; Child, Preschool; Fatty Acids; G(M1) Ganglioside; Gangliosides; Gangliosidoses; Glycolipids; Glycosphingolipids; Humans; Male; Sulfoglycosphingolipids

We studied a family with adult GM1-gangliosidosis. The proband, aged 38, had slowly progressive extrapyramidal signs with prominent dystonia, starting at about age 19. Two other patients, aged 45 and 43, had occasional slight dystonia, but led normal social lives because of mildness of their symptoms. Rectal biopsy of the proband showed histiocytic infiltration and membranous cytoplasmic bodies in the autonomic neurons. This family shows the clinical heterogeneity in adult GM1-gangliosidosis.

Topics: Adult; Biopsy; Brain; Chromatography, Thin Layer; Dystonia; Female; G(M1) Ganglioside; Gangliosides; Gangliosidoses; Histiocytes; Humans; Male; Microscopy, Electron; Middle Aged; Oligosaccharides; Pedigree; Rectum; Submucous Plexus; Tomography, X-Ray Computed

Beta-galactosidase-deficient siblings in two litters of English springer spaniel puppies showed a progressive neurological impairment, dwarfism, orbital hypertelorism, and dysostosis multiplex. An excess of GM1-ganglioside was found in the brain. Three abnormal oligosaccharides were present in samples of urine, brain, liver, and cartilage. Light microscopy of selected tissue specimens revealed cytoplasmic vacuoles in neurons, circulating blood cells, macrophages, and chondrocytes. Ultrastructural studies demonstrated that these membrane-bound vacuoles were of two types--one containing lamellated membranes and the other, finely granular material. These clinical and pathological findings are similar to those observed in human patients affected by the infantile form of GM1-gangliosidosis.

Topics: Animals; Bone Diseases, Metabolic; Dog Diseases; Dogs; Female; G(M1) Ganglioside; Gangliosidoses; Humans; Lactose Intolerance; Male; Neurons; Oligosaccharides; Pedigree; Vacuoles

The metabolism of galactosylceramide and lactosylceramide in cultured fibroblasts was studied using the lipid-loading test. These compounds were incorporated into the fibroblasts yet only small amounts of the incorporated lipids were hydrolyzed unless additional phospholipid was mixed with the glycolipid before loading. Among phospholipids, phosphatidylserine was the most effective for incorporation and hydrolysis of the glycolipids, while phosphatidylcholine inhibited the incorporation of the glycolipids. Using filtration techniques, light scattering analyses and subcellular fractionation, the particle size of glycolipid in the culture medium was found to be critically important for the incorporation of the lipids into the cells and their transportation to the lysosomes. The particle sizes of the glycolipids were decreased by mixing with phosphatidylserine. Furthermore, the negative charge in phosphatidylserine may be necessary for the glycolipid transportation into the lysosomes. In fibroblasts from patients with globoid cell leukodystrophy, 40-50% of galactosylceramide was hydrolyzed on the 4th day of culture, a time when the control fibroblasts had hydrolyzed it about 80%. This finding is in contrast with observations made on fibroblasts with other sphingolipidoses which showed near-zero degradation in corresponding substrate-loading tests. In fibroblasts from patients with either globoid cell leukodystrophy of GM1-gangliosidosis, hydrolysis of lactosylceramide was fairly normal yet somewhat lower than control values on any day of culture, thereby indicating that, in the loading tests, lactosylceramide seems to be hydrolyzed with similar levels of enzyme activities by two distinct beta-galactosidases, galactosylceramidase and GM1-ganglioside beta-galactosidase.

Topics: beta-Galactosidase; Cell Line; Cell Membrane; Cerebrosides; Fibroblasts; G(M1) Ganglioside; Galactosylceramides; Gangliosidoses; Glycosphingolipids; Humans; Hydrolysis; Lactosylceramides; Leukodystrophy, Globoid Cell; Lysosomes; Particle Size; Phosphatidylserines

Topics: Adult; beta-Galactosidase; Female; G(M1) Ganglioside; Galactosidases; Galactosylceramidase; Gangliosidoses; Genetic Variation; Humans; Male; Middle Aged; Myoclonus; Phenotype

On routine smears of blood and bone marrow of four patients with GM1 gangliosidosis type I, eosinophil granulocytes were unusually pale and contained faintly stained, unevenly spaced granules some of which were larger than normal and had abnormal ultrastructural appearance. The anomaly may represent a hitherto overlooked but easily obtainable diagnostic marker.

Topics: Eosinophils; Female; G(M1) Ganglioside; Gangliosidoses; Granulocytes; Humans; Infant

GM1 gangliosidosis is usually a pediatric disease caused by hereditary acid beta-galactosidase deficiency. There have been a few cases in adults. We saw a 51-year-old Japanese man with type 3 GM1 gangliosidosis who was manifesting dementia, dysarthria, gait disturbance, and limb rigidity. Radiologic studies showed platyspondylia, acetabular hypoplasia, and flattened femoral heads. Biochemical analysis revealed generalized acid beta-galactosidase deficiency.

Topics: beta-Galactosidase; G(M1) Ganglioside; Gangliosidoses; Humans; Male; Middle Aged

Acetylcholine metabolism was studied in cats with Gm1 gangliosidosis. Marked increases in acetylcholine synthesis (130% of controls) and K+-stimulated release of ACh (142-165% of controls) were observed in cortical and hippocampal brain slices of diseased cats. Cortical synaptosomes prepared from affected cats had significantly elevated rates of high affinity choline transport (131% of controls). These results indicate that in Gm1 gangliosidosis there is an unique disease-induced activation of cholinergic activity in the nervous system. These changes may result from disease-induced proliferation of functional cholinergic nerve endings and/or altered regulation of acetylcholine metabolism.

Topics: Acetylcholine; Animals; Brain; Cats; Cerebral Cortex; G(M1) Ganglioside; Gangliosidoses; Hippocampus; In Vitro Techniques

Two genetically distinct acid beta-galactosidases are apparently involved in the hydrolysis of galactosylceramide in fibroblasts. These beta-galactosidases were activated by different bile salts. The classical galactosylceramidase (galactosylceramidase I, EC 3.2.1.46) was activated by sodium taurocholate, while the other galactosylceramidase (galactosylceramidase II) was activated by sodium cholate. The former was genetically lacking in globoid cell leukodystrophy (GLD) and the latter in GM1 gangliosidosis. Galactosylceramidase II cross-reacted with antibody raised against purified GM1 ganglioside beta-galactosidase (EC 3.2.1.23) from the human placenta. The purified beta-galactosidase had galactosylceramidase II activity, which was competitively inhibited by GM1 ganglioside. Thus, galactosylceramidase II seems to be identical to GM1 ganglioside beta-galactosidase and lactosylceramidase II. Galactosylceramidase II had a very low affinity for galactosylsphingosine. In the galactosylceramide-loading tests using fibroblasts from patients with GLD and GM1 gangliosidosis, both cell lines hydrolyzed the incorporated galactosylceramide, with lower rates than control fibroblasts but higher than the fibroblasts from patients with I-cell disease, in which both galactosylceramidase I and II were deficient. These results indicate that galactosylceramide is hydrolyzed by two genetically distinct beta-galactosidases and explain well that galactosylsphingosine but not galactosylceramide accumulates in the brain of patients with GLD.

Topics: beta-Galactosidase; Cell Line; Cerebrosides; Chemical Precipitation; Cholic Acid; Cholic Acids; Cross Reactions; Enzyme Activation; Female; Fibroblasts; G(M1) Ganglioside; Galactosidases; Galactosylceramidase; Galactosylceramides; Gangliosidoses; Humans; Hydrogen-Ion Concentration; Immunologic Techniques; Leukodystrophy, Globoid Cell; Placenta; Pregnancy; Psychosine; Substrate Specificity; Taurocholic Acid

A new fluorogenic compound--6-hexadecanoylamino-4-methyl-umbelliferyl-beta-D-gala cto pyranoside (HMGal), a substrate for human galactocerebroside beta-D-galactosidase (HG), has been synthesized. A method for determining the HG activity based on the use of HMGal as a fluorogenic substrate has been developed. The specificity of HMGal hydrolysis by HG has been demonstrated in experiments with enzyme preparations from human skin fibroblasts and leukocytes in normally and in hereditary glycolipidosis (GM1-gangliosidosis and Krabbe's disease). The use of HMGal permits to markedly increase the sensitivity of the method used for determining the HG activity.

Topics: Chemical Phenomena; Chemistry; Chromogenic Compounds; Clinical Enzyme Tests; G(M1) Ganglioside; Galactosidases; Galactosylceramidase; Gangliosidoses; Humans; Leukodystrophy, Globoid Cell

Type III GM1-gangliosidosis is a rare hereditary storage disease caused by lack of lysosomal beta-galactosidase and characterized by a slowly progressive course, and extrapyramidal signs, but without prominent skeletal changes or visceromegaly. The storage substance was reported to be located only in the basal ganglia. There has been no detailed report on visceral lesions in type III GM1-gangliosidosis. In this report we describe a case of type III GM1-gangliosidosis, and the histochemical and ultrastructural findings from biopsied rectum. The patient was a 22-year-old female who exhibited dysarthria, gait disturbance, and generalized dystonia with rigidity. Beta-galactosidase activity in leukocytes was absent and sialidase activity in cultured fibroblasts was normal. Many histiocytes were found in biopsied rectal mucosa. Histochemical studies showed that the granules of histiocytes contained acidic glycoconjugates, beta-galactose, beta-N-acetylgalactosamine and sialic acid. Ultrastructural investigations revealed that ganglion cells of Meissner's plexus had many osmiophilic lamellar inclusions, similar to "membranous cytoplasmic bodies". These findings are crucial for the clinical diagnosis of type III GM1-gangliosidosis.

Topics: Adult; beta-Galactosidase; Biopsy; Cytoplasmic Granules; Female; G(M1) Ganglioside; Gangliosidoses; Histiocytes; Horseradish Peroxidase; Humans; Intestinal Mucosa; Lectins; Neuraminidase; Periodic Acid-Schiff Reaction; Rectum

This communication reports the clinical and biochemical results in six patients: four with mucopolysaccharidosis, one with GM1 gangliosidosis (Morquio B) and one with I-cell disease, who were treated by amniotic tissue transplantation. The sole evident clinical result was the diminishing of corneal clouding in three cases. A slight increase of beta-galactosidase activity in one patient's plasma was observed. The time of improvement was about 2 months after the transplantation and was transitory.

Topics: Amnion; Child; Child, Preschool; Female; G(M1) Ganglioside; Gangliosidoses; Humans; Male; Mucolipidoses; Mucopolysaccharidoses; Mucopolysaccharidosis I; Mucopolysaccharidosis VI

An improved method, which combined a number of published techniques, is described for the polyethylene-glycol-induced fusion of mononuclear human skin fibroblasts in the presence of phytohaemagglutinin-P and for the subsequent isolation of polynuclear cells by Ficoll gradient sedimentation. Enriched cultures contain between 60 and 75% multinucleated cells and may be maintained in culture without fetal calf serum for up to 14 days without significant overgrowth by the few contaminating mononuclear parental cells. Complementation appears not to occur between GM1 gangliosidosis and mucopolysaccharidosis, type VI B (Morquio) cell strains; this experimental observation provides support for the earlier hypothesis that the mutations for these conditions are allelic. Earlier observations that complementation does not occur between selected phenotypic variants (viz., neuronopathic forms and those without neurological involvement) of sphingomyelin storage (Niemann-Pick) disease or Gaucher's disease are confirmed.

Topics: Cell Fusion; Cells, Cultured; Enzymes; Fibroblasts; G(M1) Ganglioside; Gangliosidoses; Gaucher Disease; Genetic Complementation Test; Humans; Lysosomes; Mucopolysaccharidosis IV; Niemann-Pick Diseases

Gangliosidoses are very rare neurological diseases based on specific enzyme defects. They constitute models for the disruption of specific metabolic pathways and cellular functions with the ultimate consequence of manifest clinical symptoms. The investigation of the various steps involved in the generation of a given syndrome can therefore lead to a more profound understanding of the cell biology of the nervous system. In the present synopsis we try to briefly summarize some aspects of the present knowledge of pathophysiological mechanisms in GM1- and GM2-gangliosidoses.

Topics: beta-Galactosidase; beta-N-Acetylhexosaminidases; Brain; Child; G(M1) Ganglioside; G(M2) Activator Protein; G(M2) Ganglioside; Gangliosides; Gangliosidoses; Glycoproteins; Hexosaminidases; Humans; Lysosomes; Nerve Degeneration; Protein Deficiency; Proteins; Saposins; Sphingolipid Activator Proteins; Synaptic Transmission

Neurophysiological studies (EEG, ERG, VEP and BAEP) have been carried out on a total of fifty-four patients (fourty-five GM2 and nine GM1 gangliosidosis) at various stages of the disease process. In infantile GM2 gangliosidosis, the EEG was midly abnormal from an early age but by the age of one year there was a rapid and progressive deterioration. EEG changes in late onset GM2 gangliosidosis were very variable and unrelated to age or enzyme defect. In both Type 1 and Type 2 GM1 gangliosidosis there was a progressive deterioration of the EEG. Paroxysmal features were not prominent in any of the gangliosidoses, despite the occurrence of seizures. The ERG remained normal in both GM2 and GM1 patients. In the infantile GM2 patients there was progressive loss of the VEP between nine and fifteen months of age but the timing of VEP changes were more variable in all the other groups. Evidence of brainstem dysfunction was found in one of the two TSD patients tested. The combined neurophysiological features appear to be characteristic for each group of gangliosidosis and differ from other neurometabolic disorders of childhood.

Topics: Arousal; Brain; Child; Child, Preschool; Electroencephalography; Electroretinography; Evoked Potentials; G(M1) Ganglioside; G(M2) Ganglioside; Gangliosides; Gangliosidoses; Humans; Infant; Sandhoff Disease; Synaptic Transmission; Tay-Sachs Disease

Topics: Axons; Cell Line; Child; G(M1) Ganglioside; G(M2) Ganglioside; Gangliosides; Gangliosidoses; Humans; Motor Endplate; Motor Neurons; Muscles; Nerve Degeneration; Nerve Growth Factors; Nerve Regeneration; Neuromuscular Junction; Neurons; Peripheral Nerves; Synaptic Transmission

GM1 and GM2 gangliosidoses are progressive neurodegenerative diseases which accumulate intralysosomal gangliosides--and to a lesser extent oligosaccharides--chiefly in the central and peripheral nervous system owing to deficiencies of beta-galactosidase and hexosaminidases A or/and B, respectively. This intralysosomal "storage" in neuronal pericarya and their processes, and subsequent loss of such nerve cells provide the background for clinical symptoms of the central nervous system and the retina, while involvement of the peripheral nervous system and the visceral organs largely remains free of clinical findings. The morphological involvement of the latter organs is widespread though varying, thus allowing morphological investigations of lymphocytes, skin, or rectum for morphological diagnosis and as a screening procedure.

Topics: Astrocytes; Brain; Child; Dendrites; G(M1) Ganglioside; G(M2) Ganglioside; Gangliosides; Gangliosidoses; Humans; Inclusion Bodies; Lysosomes; Microscopy, Electron; Nerve Degeneration; Neurons; Peripheral Nerves; Sandhoff Disease; Spinal Cord; Synaptic Membranes; Tay-Sachs Disease; Vacuoles

An increasing focus on the mechanism of synaptic neurochemistry in pediatric neurology, may lead to a better understanding of the pathophysiology of many disorders and result in a more rational approach to their pharmacotherapy. With the burgeoning list of putative neurotransmitters in brain, and the growing evidence of co-localization of many of these neurotransmitters, chemical neurotransmission likely involves a higher degree of complexity than appreciated heretofore. The potential role of neurotransmitter dysfunction in the pathophysiology of neurologic and behavior disorders of children, should not be considered as restricted to those disorders that involve selective neuronal loss, but may encompass a much wider spectrum of syndromes due to metabolic abnormalities, as well as disturbances of the finer features of chemical neurotransmission.

Topics: Afferent Pathways; Animals; Brain Damage, Chronic; Cats; Central Nervous System; Cerebral Cortex; Chemical Phenomena; Chemistry; Child; Child, Preschool; G(M1) Ganglioside; gamma-Aminobutyric Acid; Gangliosidoses; Humans; Mental Disorders; Methylazoxymethanol Acetate; Nerve Tissue; Nervous System Diseases; Neurons; Neurotransmitter Agents; Rats; Sympathetic Nervous System; Synapses; Tourette Syndrome

Topics: beta-Galactosidase; beta-N-Acetylhexosaminidases; Brain; Child; Fibroblasts; G(M1) Ganglioside; G(M2) Activator Protein; G(M2) Ganglioside; Gangliosides; Gangliosidoses; Glycoproteins; Hexosaminidases; Humans; Lysosomes; Mutation; Protein Deficiency; Proteins; Sandhoff Disease; Saposins; Sphingolipid Activator Proteins; Tay-Sachs Disease

Abnormal membranous cytoplasmic inclusions were found in the retinal ganglion cells of two fetuses with gangliosidosis. One was a documented case of incipient Tay-Sachs disease (Gm2) and the other a case of generalized gangliosidosis (Gm1). Both specimens were obtained iatrogenically in the 20th to 21st week of gestation after amniocentesis had indicated the enzyme deficiency.

Topics: Amniocentesis; Female; G(M1) Ganglioside; Gangliosides; Gangliosidoses; Gestational Age; Humans; Inclusion Bodies; Microscopy, Electron; Pregnancy; Retina; Retinal Ganglion Cells; Tay-Sachs Disease

The nature of the molecular defect resulting in the beta-galactosidase deficiency in different forms of GM1-gangliosidosis and mucopolysaccharidosis IV B (Morquio B syndrome) was investigated. Normal and mutant cultured skin fibroblasts were labeled in vivo with [3H]leucine and immunoprecipitation studies with human anti-beta-galactosidase antiserum were performed, followed by polyacrylamide gel electrophoresis and fluorography. In Morquio B syndrome, the mutation does not interfere with the normal processing and intralysosomal aggregation of beta-galactosidase. In cells from infantile and adult GM1-gangliosidosis, 85-kDa precursor beta-galactosidase was found to be synthesized normally but more than 90% of the enzyme was subsequently degraded at one of the early steps in posttranslational processing. The residual 5-10% beta-galactosidase activity in adult GM1-gangliosidosis is 64-kDa mature lysosomal enzyme with normal catalytic properties but with a reduced ability of the monomeric form to aggregate into high molecular weight multimers. Knowledge of the exact nature of the molecular defect underlying beta-galactosidase deficiency in man may lead to a better understanding of the clinical and pathological heterogeneity among patients with different types of GM1-gangliosidosis and Morquio B syndrome.

Topics: beta-Galactosidase; Fibroblasts; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Humans; Lysosomes; Mucopolysaccharidosis IV; Mutation; Skin

A case of type 1 GM1 gangliosidosis, also called Norman-Landing disease is reported. The patient' was a nine months old boy who presented psychomotor retardation since birth, coarse facies, hepatomegaly and macular cherry red spot. Roentgenographic findings were those of dysostosis multiplex. Bone marrow smear showed type 1 Gasser's cells, as it occurs in the storage diseases. The infant presented a severe B-galactosidase deficiency and died at the age of ten months. Recent advances in pathogenesis, diagnosis and future therapy are discussed.

Topics: beta-Galactosidase; G(M1) Ganglioside; Gangliosidoses; Genetic Carrier Screening; Humans; Infant; Isoenzymes; Male; Prenatal Diagnosis

The results of the diagnostic application of first trimester trophoblast sampling in 100 pregnancies are reported in detail. Further improvement of the method for routine, direct chromosome analysis resulted in a technique which proved to be fast, simple, and efficient. We found that short-term incubation of villi permits the application of many experimental methods, such as visualization of sister chromatid exchanges and bromodeoxyuridine (BrdU) incorporation. Fetal karyotyping was successful in each of the 96 pregnancies in which fetal material was obtained from a total of 98 fetuses. There were 42 males and 56 females, and an abnormal chromosome constitution was found in 12 cases. Two trisomic fetuses were found among the eight pregnancies at risk for Duchenne muscular dystrophy, and this indicates that fetal sexing (which is achieved with our method in two hours) should not be performed without chromosome visualization. The results indicate a risk of 8% of an abnormal fetus for mothers aged 35 years or more, while the risk of failure of sampling and of spontaneous abortion after villi sampling were 4 and 6%, respectively. Enzyme determinations were performed in three pregnancies at risk for gangliosidosis GM1, Niemann-Pick disease, and Hurler syndrome. In this last case inconsistency between the results of the assay of iduronidase on chorionic villi and amniotic fluid cells was found. This unexplained error indicates the need for extensive characterisation in chorionic villi of the series of enzymes involved in metabolic diseases.

Topics: Adult; Chorionic Villi; Chromosome Aberrations; Chromosomes, Human, 13-15; Chromosomes, Human, 16-18; Clinical Enzyme Tests; Female; G(M1) Ganglioside; Gangliosidoses; Humans; Karyotyping; Male; Maternal Age; Niemann-Pick Diseases; Placenta; Pregnancy; Pregnancy Trimester, First; Pregnancy, High-Risk; Prenatal Diagnosis; Trophoblasts

Biochemical analyses of the samples of GM1-gangliosidosis Type 1, Type 2, and Chronic type at autopsy showed that GM1 and asialo-GM1 are markedly increased in the whole cerebral tissues of patients with Type 1 and Type 2, but mainly in the basal ganglia including caudate nucleus and putamen in the Chronic Type as already reported by Kobayashi and Suzuki in 1981. On the other hand, the finding that the Type 1 visceral organs contained unusual glycolipids rather than gangliosides was contrasted to those of Type 2 which seemed less abnormal. The unusual glycolipids included particularly fucolipid I and II consisting of glucose:galactose:N-acetylglucosamine:fucose (1:2:1:1) and (1:3:2:2), respectively. The chemical structure of oligosaccharide moiety of fucolipid I may be identical with Lacto-N-fucopentaose II or Lacto-N-fucopentaose III. The fucolipid II may be proposed to be Lacto-N-difucooctanose.

Topics: Brain Chemistry; Cerebrosides; Child, Preschool; Chromatography, Thin Layer; Female; G(M1) Ganglioside; Gangliosides; Gangliosidoses; Glycolipids; Humans; Kidney; Lipids; Liver; Spleen

Previous studies using the lectin RCA-I from Ricinus communis have indicated that several lysosomal enzymes in the fibroblasts of patients deficient in beta-galactosidase carry excess terminal galactose. Electrophoretic studies have shown that the same enzymes and the non-lysosomal adenosine deaminase also show excess terminal sialic acid in patients deficient in sialidase. In this paper we confirm, using Jack-bean beta-galactosidase, that the binding to RCA-I of the purified N-acetyl-beta-D-hexosaminidase from a patient with GM1 gangliosidosis depends on a terminal beta-linked galactose. We provide evidence, using bacterial sialidase and measuring the binding to RCA-I, for excess subterminal galactose on the enzymes of patients deficient in sialidase. We also show that adenosine deaminase from the fibroblasts of patients deficient in beta-galactosidase has increased binding to RCA-I. These observations suggest that in healthy individuals the carbohydrate structure of the precursors of lysosomal enzymes and possibly some other glycoproteins also includes extended carbohydrate side chains with terminal sialic acid and subterminal galactose, and that the mature enzyme extracted from tissues is the product of degradation.

Topics: Acetylglucosaminidase; Adenosine Deaminase; Adolescent; Adult; beta-Galactosidase; Carbohydrate Metabolism; Cell Line; Child; Chromatography, Affinity; Female; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Glycoproteins; Humans; Lysosomes; Male; Middle Aged; Neuraminidase

The enzymatic diagnosis of GM1 gangliosidosis, including the diagnosis of heterozygosity, requires a microassay of GM1 ganglioside beta-galactosidase activity in lymphocytes and cultured skin fibroblasts. We have adopted high-performance liquid chromatography (HPLC) to the assay of this enzyme and can measure the activity in crude samples fluorometrically. Reaction conditions were examined to determine those optimal for the assay of GM1 ganglioside beta-galactosidase activity in lymphocyte and skin fibroblast homogenates. Under these optimal conditions, reduced enzymatic activities could be detected in lymphocytes and cultured skin fibroblasts from three patients with GM1 gangliosidosis. Thus, this assay can be used for the diagnosis, rather than the usual assays employing radioactive or artificial substrates.

Topics: Adult; beta-Galactosidase; Chromatography, High Pressure Liquid; Detergents; Fibroblasts; G(M1) Ganglioside; Galactosidases; Gangliosides; Gangliosidoses; Humans; Hydrogen-Ion Concentration; Lymphocytes; NAD

A patient with combined deficiency of sialidase and beta-galactosidase is described. This now 39-year-old man, who is of Japanese origin, showed gradually progressive clinical features from the age of six years. Many of these features are commonly found in sialidosis type 2 or in GM1-gangliosidosis. Both sialidase and beta-galactosidase activities were deficient in leucocytes and cultured fibroblasts. Leucocytes of his mother showed activities of both enzymes in the lower limit of the control range. Morphologically, the pattern of storage products in a skin biopsy resembled in many respects that seen in GM1-gangliosidosis. Moreover, storage products which could be typical of sialidosis were also observed. Since the patient showed angiokeratomata, the morphological findings were compared with those specific to Fabry's disease, but no similarities were found. An enzymological diagnosis of the disease is most reliable on cultured fibroblasts, discriminating it from sialidosis type 2 and GM1-gangliosidosis. In view of recent findings, leucocytes seem to be less suitable for the establishment of the diagnosis galactosialidosis.

Topics: Adult; Axons; Fibroblasts; G(M1) Ganglioside; Gangliosidoses; Humans; Lactose Intolerance; Leukocytes; Male; Neuraminidase; Skin; Vacuoles

Authors present the case of a child, daughter of non related parents with neurologic progressive affectation, retina and visceral implication with certain pseudogargolic clinical aspect without near familiar antecedents suggestive of this disease. Biochemistry and histologic studies revealed a B-galactosidase enzyme deficiency and lipidic intracellular increase in different viscera. Biochemistry, hystological and clinical aspects exposed are fundamentally differential of another causes of pseudo-Hurler syndrome.

Topics: beta-Galactosidase; beta-Glucosidase; Female; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Humans; Infant, Newborn; Sphingolipidoses; Sphingomyelin Phosphodiesterase

Sphingolipid activator protein-1 (SAP-1) is a glycoprotein found in human tissue extracts that stimulates the enzymatic hydrolysis of at least two glycosphingolipids, including GM1 ganglioside and sulfatide. The ability of purified SAP-1 to stimulate GM1 ganglioside hydrolysis by extracts of cultured fibroblasts from patients with beta-galactosidase deficiency was examined, and all patients had a pronounced deficiency (under 10% of control). Using monospecific antibodies against SAP-1, the concentration was determined in cultured fibroblasts by rocket immunoelectrophoresis. Extracts from 15 control cell lines were found to have 0.72 +/- 0.24 micrograms cross-reactive material/mg protein, while cell extracts from 8 patients with GM1 gangliosidosis involving mental retardation were found to have 1.08 +/- 0.17, which is significantly elevated. When the fibroblast extracts were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by electroblotting, multiple bands were observed. Controls were found to have two major bands with estimated molecular weights of 9000 and 9500, and a minor band at 7800. Extracts from patients with GM1 gangliosidosis were found to have multiple bands ranging upward to 13,000. Extracts from patients with the most severe clinical types of GM1 gangliosidosis had almost exclusively high-molecular-weight forms (molecular weights above 10,000). Treatment of SAP-1 from control liver with endoglycosidase D caused a decrease in the Mr 9500 band and increased in the Mr 7800 band. When SAP-1 from GM1 gangliosidosis liver was treated sequentially with neuraminidase, beta-galactosidase, and endoglycosidase D, almost all of it was converted to the forms found in control human liver.

Topics: Cells, Cultured; Electrophoresis, Polyacrylamide Gel; Fibroblasts; G(M1) Ganglioside; Gangliosidoses; Glycoproteins; Humans; Hydrolysis; Immunoelectrophoresis; Lactose Intolerance; Metabolism, Inborn Errors; Molecular Weight; Proteins; Saposins; Sphingolipid Activator Proteins

Topics: Female; G(M1) Ganglioside; Gangliosides; Gangliosidoses; Humans; Infant; Tomography, X-Ray Computed

Topics: Adult; beta-Galactosidase; Female; Fibroblasts; G(M1) Ganglioside; Gangliosides; Gangliosidoses; Humans; Infant; Lactose Intolerance; Placenta; Pregnancy; Reference Values

The clinical and pathological manifestations of a case of juvenile GM1 gangliosidosis are presented and the pathological findings compared with those previously reported for GM1 gangliosidosis in man and in animal models. The most striking finding in the present case was the marked degeneration of the retinal ganglion cell and nerve fiber layers. Although such extensive ganglion cell loss was not observed in any of the other cases reviewed, the presence of multimembranous inclusion bodies in retinal ganglion cells strongly suggests that the pathological process was similar in all cases. Much remains to be learned about the function of gangliosides in the healthy retina and about the pathophysiological consequences of deranged ganglioside metabolism. The many parallels, including those observed in pathological studies, between the human and animal forms of GM1 gangliosidosis allow an optimistic appraisal of the value of further research using the animal models.

Topics: Animals; Atrophy; Cats; Cattle; Conjunctiva; Cornea; Disease Models, Animal; Dogs; Epithelium; Eye; Eye Diseases; Female; G(M1) Ganglioside; Gangliosidoses; Humans; Infant; Retina; Retinal Ganglion Cells

Glycosphingolipids from the liver, kidney, and spleen of a patient with type 1 II3-N-acetylneuraminosylgangliotetraosylceramide (GM1)-gangliosidosis were quantitatively analyzed. It was noted that large amounts of unusual glycosphingolipids other than GM1 ganglioside or gangliotetrasylceramide accumulated in the liver of the patient. Particularly, the prominent accumulation of III3-alpha-fucosylneolactotetraosylceramide, galactosylceramide I3-sulfate and cholesterol sulfate was observed in addition to a small but significant increase of galabiosylceramide and neolacto-or lactotetraosylceramide. None of these lipids except cholesterol sulfate can be detected in normal liver. None of the lipids accumulated in the liver can be the direct substrates for acid beta-galactosidase which is deficient in the patient. Thus, it was suggested that secondary effects due to the defect in acid beta-galactosidase might cause the abnormal accumulation of various lipids in the liver.

Topics: Cholesterol; Chromatography, Thin Layer; Fatty Acids; Female; G(M1) Ganglioside; Gangliosides; Gangliosidoses; Glycosphingolipids; Humans; Kidney; Liver; Spectrophotometry, Infrared; Spleen; Tissue Distribution

The GM1 gangliosidoses are clinically characterized by the combination of a degenerative process in the brain and of storage phenomena in extra-neural tissues, particularly in bones and visceral organs. Phenotypic variability is pronounced. "Classical" types, according to the age at onset, are infantile ("generalized"), juvenile, and adult forms. In rare variants, the degenerative process may be restricted to the basal ganglia and cause dystonia musculorum deformans, or it may cause infantile cardiomyopathy. Much of this variability may be explained by variable residual activities of the deficient beta-galactosidase towards various substrates.

Topics: Adolescent; Adult; beta-Galactosidase; Brain; Child; Child, Preschool; G(M1) Ganglioside; Gangliosidoses; Humans; Infant; Infant, Newborn; Neurons; Phenotype

Sphingomyelinase activity of cultivated skin fibroblast extracts from normal individuals was resolved by chromatofocusing in the pH range 8-5 into three major components with pI's of 7.3, 6.3 and 5.9, respectively. Chromatofocusing proved a more efficient and reproducible separation technique than preparative flat-bed isoelectric focusing and it gave a constant profile even when detergent concentration varied. In skin fibroblasts from five patients with Niemann-Pick disease type C, a varying degree of reduction in the proportion of the 7.3 peak was observed. In a patient with clinical features of Niemann-Pick disease type C, the finding of such a profile would thus be a good argument for the diagnosis, but it is not pathognomonic as we found similar changes in two cases with GM1-gangliosidosis, while some cases of Niemann-Pick disease type C have borderline normal profiles. These results challenge the concept of a specific sphingomyelinase isoenzyme deficiency as the basic defect in Niemann-Pick disease type C.

Topics: Fibroblasts; G(M1) Ganglioside; Gangliosidoses; Humans; Hydrogen-Ion Concentration; Isoelectric Focusing; Isoelectric Point; Niemann-Pick Diseases; Phosphoric Diester Hydrolases; Skin; Sphingomyelin Phosphodiesterase

An improved, rapid, and sensitive method for the biochemical diagnosis of GM1 gangliosidosis based on the detection and quantification of urinary galactosyl-oligosaccharides with high performance liquid chromatography was developed. The oligosaccharides, in 50-100 microliters of urine, were converted to radioactively labeled oligosaccharide-alditols with NaB3H4 and fractionated on commercial silica-amine bonded, high performance liquid chromatography columns. Delineation between infantile, juvenile, and adult onset subtypes of GM1 gangliosidosis was possible by analysis of the levels of the excreted oligosaccharides and their characteristic elution profile. Infantile and juvenile patients contain identical numbers of oligosaccharide fractions (13 resolved components) but can be distinguished by 3-10-fold lower levels of oligosaccharides in juvenile patients and, in some cases by a disproportionately lower concentration of high molecular weight compounds. Adult onset patients were distinguished by substantially lower concentrations of urinary oligosaccharides, 130-180-fold below those in infantile patients, and the apparent absence of high molecular weight oligosaccharides.

Topics: Adult; Age Factors; Animals; Child; Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Dogs; G(M1) Ganglioside; Gangliosidoses; Humans; Infant; Liver; Male; Oligosaccharides

The morphopathologic abnormalities characterizing the gangliosidoses, e.g., meganeurites and aberrant secondary neurites with associated dendritic spines and synapses, show pronounced regional variation. Because gangliosides are thought to be the causative agents, we undertook to detect possible variation in concentration or composition that would correlate with those morphologic findings. A gradient in total ganglioside and GM1 concentration was found corresponding to cerebral cortex greater than caudate = thalamus greater than cerebellum, similar to the morphologic gradient. In addition, the fatty acid compositions were variable, the proportion of docosanoate (22:0) following the same gradient. The possibility that these variations in ganglioside content and composition might influence the growth of aberrant neurites and related structures is discussed.

Topics: Animals; Brain; Brain Chemistry; Cats; G(M1) Ganglioside; Gangliosides; Gangliosidoses; Humans

Prenatal diagnosis of infantile GM1 gangliosidosis was accomplished by analyzing the galactosyl-oligosaccharides accumulating in amniotic fluid with high-performance liquid chromatography (HPLC) at 14 weeks gestation. The pattern of amniotic oligosaccharides was nearly identical with that in neonatal urine in GM1 gangliosidosis but the concentrations were about one-fiftieth of that in urine, necessitating a highly sensitive assay.

Topics: Amniotic Fluid; Chromatography, High Pressure Liquid; Female; G(M1) Ganglioside; Gangliosidoses; Humans; Oligosaccharides; Pregnancy; Prenatal Diagnosis

Gangliosides were previously reported to induce neuritogenesis in primary neuronal cultures and in some neurally derived cell lines. Because isolated gangliosides usually contain variable quantities of peptides, we investigated the possibility that neurite-stimulating activity could be caused by these contaminants. Ganglioside preparations from bovine brain and other sources were subjected to a three-step purification procedure that eliminated at least 95% of the contaminating peptides. These purified preparations retained their capacity to induce extensive neurite growth in neuro-2A murine neuroblastoma. Proteolytic digestion and a number of additional procedures were used to reduce residual contamination further without loss of activity. Several crude ganglioside samples had negative effects on neurite development until freed of their inhibitory factors, which were derived from the tissue and/or introduced during laboratory operations. This was particularly evident for bovine white matter gangliosides whose activity increased in proportion to peptide removal. When carefully purified, virtually all of 11 different gangliosides tested were highly active, with the possible exception of GM4, which demonstrated only moderate activity in a limited number of tests. All of the neutral glycolipids tested, as well as sulfatides and free sialic acid, were inactive.

Topics: Animals; Cats; Cattle; Cell Division; G(M1) Ganglioside; Gangliosides; Gangliosidoses; Glycolipids; Humans; Neurons

The nervous system of a 22-year-old fetus with GM1-gangliosidosis type 1 was studied by electron microscopy. The tissues thus examined were the cerebral cortex at the parietal region, the cerebellum, the thoracic spinal cord, the Auerbach's myenteric plexus in the large intestine and the radial nerve fibers. In the cerebral cortex, membrane-bound vacuoles, which occasionally contained stacks of fine fibrils, were observed in the large young neurons in the deeper part of the cortical plate. The neurons in the other part of the cerebral cortex carried no storage materials. In the cerebellum, the membrane-bound vacuoles with stacks of fine fibrils were seen only in the Purkinje cells. The neurons in the spinal cord also contained several zebra-like bodies and the above membrane-bound vacuoles. As for the peripheral nervous system (PNS), neurons in the Auerbach's myenteric plexus carried membranous cytoplasmic bodies and zebra-like bodies. Some of the axons in the radial nerve fibers also contained a lot of pleomorphic electron-dense bodies and a few membranous cytoplasmic ones. These results show that the accumulation of storage materials is started in the large neurons which are produced in the early stage of neurogenesis in the central nervous system (CNS). Additionally, the observed membrane-bound vacuoles are considered to be structures which occur before the membranous cytoplasmic bodies and/or the zebra-like bodies. It is also elucidated that the PNS is affected earlier than the cerebral and cerebellar cortices and thoracic spinal cord.

Topics: Cerebellum; Cerebral Cortex; Fetus; G(M1) Ganglioside; Gangliosidoses; Humans; Microscopy, Electron; Myenteric Plexus; Nervous System; Parietal Lobe; Radial Nerve; Spinal Cord; Vacuoles

Urine samples from patients with different types of glycoprotein storage disease were chromatographed by gel filtration and the fractions analysed for sialic acid. Patients with mucolipidoses I and II excreted the largest amounts of bound sialic acid. One patient with GM1 gangliosidosis showed an abnormal level of sialyloligosaccharide excretion. Other patients showed normal results. With the present method mucolipidoses I and II, together with GM1 gangliosidosis, are readily distinguished from other possible oligosaccharidurias.

Topics: G(M1) Ganglioside; Gangliosidoses; Humans; Lactose Intolerance; Mucolipidoses; Mucopolysaccharidoses; N-Acetylneuraminic Acid; Neuraminidase; Oligosaccharides; Sialic Acids

Assays for synaptosomal high-affinity uptake activity (glutamate, gamma-aminobutyric acid, norepinephrine), neurotransmitter synthesizing enzymes (choline acetyltransferase, glutamate decarboxylase, tyrosine hydroxylase), and endogenous neurotransmitters were performed in cats with advanced inherited GM1 gangliosidosis. A significant reduction in uptake activity, ranging from 24 to 77% of control, was demonstrated in motor, occipital, and cerebellar brain regions. This reduction was unassociated with comparable alterations in neurotransmitter levels or synthesizing enzyme activity. We hypothesize that the defect of neurotransmitter inactivation is part of an overall abnormality of synaptic membrane function that could contribute to the neurological symptoms seen in the hereditary gangliosidoses.

Topics: Animals; Brain; Cats; Cerebellum; Disease Models, Animal; Frontal Lobe; G(M1) Ganglioside; gamma-Aminobutyric Acid; Gangliosidoses; Glutamates; Glutamic Acid; Humans; Neurotransmitter Agents; Norepinephrine; Occipital Lobe; Synaptosomes

Topics: beta-Galactosidase; Chromatography, Paper; Electrophoresis; Fibroblasts; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Glycosaminoglycans; Humans; Keratan Sulfate; Sulfates; Trisaccharides

Topics: Animals; Cats; Cerebellum; Cerebral Cortex; Freeze Fracturing; G(M1) Ganglioside; Gangliosidoses; Hippocampus; Humans; Inclusion Bodies; Microscopy, Electron; Neurons; Synaptic Membranes

A homologous series of structurally related, high molecular weight oligosaccharides have been isolated and purified from the livers of a mixed breed of Beagle dogs affected with GM1 gangliosidosis. Five individual oligosaccharide fractions were purified by charcoal chromatography, preparative silicic acid thin layer chromatography, and gel filtration chromatography. Molecular size determinations revealed that these oligosaccharides contained 6, 9, 11, and 13 sugar residues, respectively. Detailed structure analysis was carried out on the most abundant fractions, oligosaccharides 1,2 and 3 (OS 1,2 and OS 3) using permethylation analysis and 360-MHz proton magnetic resonance spectroscopy coupled with sequential exoglycosidase degradation. OS 1,2 was a mixture of two linear isomeric hexasaccharides and OS 3 was a nonasaccharide containing a bianntenary branched mannosyl core. The proposed structures are: (formula see text) These compounds are nearly identical with the oligosaccharides stored in human GM1 gangliosidosis liver but they differ from the human compounds uniquely since they contain 2 GlcNAc residues at the reducing terminus instead of 1, suggesting that there may be significant differences in glycoprotein metabolism or structure between mammalian species.

Topics: Animals; Carbohydrate Conformation; Carbohydrate Sequence; Dog Diseases; Dogs; G(M1) Ganglioside; Gangliosidoses; Humans; Liver; Magnetic Resonance Spectroscopy; Molecular Weight; Oligosaccharides

Topics: Animals; beta-Galactosidase; Cattle; Electrophoresis, Polyacrylamide Gel; Endocytosis; Fibroblasts; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Haplorhini; Humans; Male; Neuraminidase; Testis; Time Factors

Topics: Ascites; Female; G(M1) Ganglioside; Gangliosidoses; Humans; Infant, Newborn; Intestinal Mucosa; Liver; Male; Rectum

An 11-year-old boy with type 2 GM1-gangliosidosis was presented, providing further evidence for the clinical and biochemical heterogeneity of the disease. The patient had several characteristics of type 2 GM1-gangliosidosis, but was different from so-called typical type 2 GM1-gangliosidosis from the point of view of survival and the degree of GM1-ganglioside accumulation. GM1-gangliosidosis was diagnosed by absence of beta-galactosidase activity in leukocytes and the parents had the enzyme levels of heterozygotes. However, the amount of the brain GM1-ganglioside was accumulated to a less degree in comparison with that of typical type 2 GM1-gangliosidosis, though the activity of GM1-beta-galactosidase in the brain was deficient to the same degree as in the typical case.

Topics: beta-Galactosidase; Child; G(M1) Ganglioside; Gangliosidoses; Humans; Lactose Intolerance; Leukocytes; Male; Prognosis

Topics: Enzyme Activation; G(M1) Ganglioside; Galactosidases; Galactosylceramidase; Gangliosidoses; Glycoproteins; Glycosphingolipids; Humans; Hydrolysis; In Vitro Techniques; Liver; Molecular Weight; Proteins; Saposins; Sphingolipid Activator Proteins

Kinetic studies of 4-methylumbelliferyl neuraminidase activity were carried out in cultured skin fibroblasts from patients with various disorders of neuraminidase deficiency. Cell extracts from two patients with dysmorphic type sialidosis of infantile onset, with isolated deficiency of neuraminidase activity, and three patients with dysmorphic type sialidosis of juvenile onset, with combined deficiency of neuraminidase and beta-galactosidase activities, demonstrated 7-12 times higher apparent Km values than those of normal controls (1.0-1.5 mmol/l as compared with 0.12-0.15 mmol/l). The apparent Ki values for N-acetylneuraminic acid and colominic acid were also increased in the dysmorphic type (7-15 and 7-11 times the normal values, respectively). In contrast, in the normomorphic type, normal apparent Km and Ki values were found for 4-methylumbelliferyl neuraminidase activity in fibroblasts from one patient with isolated neuraminidase deficiency and two patients with combined deficiency of neuraminidase and beta-galactosidase. The altered kinetics in the dysmorphic cases indicates a primary defect in neuraminidase with a secondary deficiency of beta-galactosidase in patients with combined deficiency. It is not clear if the primary defect in the normomorphic cases involves a defect in neuraminidase other than a Km defect or if neuraminidase or both neuraminidase and beta-galactosidase deficiencies are secondary to another defect as yet undetermined.

Topics: Cell Line; Fibroblasts; G(M1) Ganglioside; Gangliosidoses; Humans; Hymecromone; Kinetics; Lactose Intolerance; Mucolipidoses; Neuraminidase

Topics: beta-Galactosidase; beta-Glucosidase; Cell Membrane; Chromatography, Gel; G(M1) Ganglioside; Gangliosidoses; Gaucher Disease; Glucosidases; Glucosylceramidase; Hot Temperature; Humans; Isoelectric Focusing; Spleen; Substrate Specificity

The ultrastructural and biochemical features of canine GM1 gangliosidosis were studied. beta-Galactosidase activity assayed using both skin fibroblast tissue culture strains and fresh skin revealed enzyme activities in three groups (normals, heterozygotes, and homozygotes) corresponding to an autosomal recessive inheritance. The concentration of ganglioside GM1 was greatly increased in cerebral gray matter and kidney. A striking elevation of tissue oligosaccharides was found in liver, kidney, and spleen. Most neurons in the cerebral cortex and deep gray matter were filled by spherical lamellated inclusions. Hepatocytes contained vacuoles with an amorphous granular material which may correspond to the accumulation of galactose-oligosaccharides determined chemically. The disease in dogs has features similar to both the infantile and juvenile form of human GM1 gangliosidosis.

Topics: Animals; Brain; Brain Chemistry; Disease Models, Animal; Dog Diseases; Dogs; Female; G(M1) Ganglioside; Gangliosidoses; Heterozygote; Homozygote; Humans; Liver; Male

Topics: Female; G(M1) Ganglioside; Gangliosidoses; Humans; Infant; Lactose Intolerance; Male; Microscopy, Electron; Pedigree; Skin; Tomography, X-Ray Computed

An infant with GM1 gangliosidosis was found to have an eruption at birth consisting of extensive and unusual slate blue macules resembling mongolian spots. All areas of skin were involved except face, scalp, palms, and soles. A biopsy of a macule obtained at 5 months of age demonstrated melanocytic cells in the dermis consistent with monogolian spot but also a perivascular histiocytic infiltrate. At 8 months of age, absence of beta-galactosidase activity was documented in both peripheral leukocytes and skin fibroblasts confirming the diagnosis of GM1 gangliosidosis. The dermal histiocytic cells noted on skin biopsy were interpreted as a manifestation of this storage disease. The coexistence of the hyperpigmented lesions and the heritable enzyme defect was believed to be coincidental.

Topics: beta-Galactosidase; G(M1) Ganglioside; Gangliosidoses; Humans; Infant; Lactose Intolerance; Nevus, Pigmented; Skin; Skin Neoplasms

Infantile, juvenile, and adult forms of GM1 gangliosidosis have been well characterized. Certain genetic and biochemical studies have suggested that the phenotypic variation found in GM1 gangliosidosis results from different allelic mutations affecting the GM1 ganglioside beta-galactosidase locus and that different combinations of these mutations accounts for the clinical heterogeneity of this illness. A family in which both the infantile and juvenile forms of GM1 gangliosidosis occurred, the children sharing a common mutation of their acid beta-galactosidase activity, supports the allelic nature of these different clinical forms of the disease. From the observations made in this unique family, additional phenotypes of GM1 gangliosidosis might be anticipated.

Topics: Alleles; beta-Galactosidase; Female; G(M1) Ganglioside; Gangliosides; Gangliosidoses; Genetic Variation; Humans; Male; Pedigree; Phenotype

Acid beta-galactosidase activity can be separated into multiple molecular forms by isoelectric focusing on cellulose acetate membranes. The residual acid beta-galactosidase in the juvenile form of GM1 gangliosidosis has three bands of enzyme activity with an apparent isoelectric pH (pI) range from 4.9 to 5.2, whereas that in the infantile form has a single band with an apparent pI of 5.2. Separation of residual acid beta-galactosidase into multiple molecular forms by analytical isoelectric focusing demonstrates enzymatic differences that can be correlated with the allelic mutations that affect the GM1 ganglioside beta-galactosidase locus.

Topics: beta-Galactosidase; Culture Techniques; Electrophoresis, Cellulose Acetate; Female; Fibroblasts; G(M1) Ganglioside; Galactosidases; Gangliosides; Gangliosidoses; Genetic Variation; Humans; Male; Protein Biosynthesis; Skin

Oligosaccharide patterns obtained by gel filtration of the urine of GM1-gangliosidosis Type 1 patients are quite different from those of GM1-gangliosidosis Type 2. By studies of oligosaccharides in the four major peaks obtained from the Type 1 subgroup using sequential exoglycosidase digestion, methylation analysis, and periodate oxidation, the structures of 15 oligosaccharides: Gal beta 1 leads to 4GlcNAc beta 1 leads to 2Man alpha 1 leads to 3Man beta 1 leads to 4GlcNAc, Gal beta 1 leads to 4GlcNAc beta 1 leads to 2Man alpha 1 leads to 6Man beta 1 leads to 4GlcNAc, Man alpha 1 leads to 6(Gal beta 1 leads to 4GlcNAc beta 1 leads to 2Man alpha 1 leads to 3)Man beta 1 leads to 4GlcNAc, Gal beta 1 leads to 4GlcNAc beta 1 leads to 2Man alpha 1 leads to 6(Gal beta 1 leads to 4GlcNAc beta 1 leads to 2Man alpha 1 leads to 3)Man beta 1 leads to 4GlcNAc, Gal beta 1 leads to 4GlcNAc beta 1 leads to 2Man alpha 1 leads to 6[Gal beta 1 leads to 4GlcNAc beta 1 leads to 4(Gal beta 1 leads to 4GlcNAc beta 1 leads to 2)Man alpha 1 leads to 3]Man beta 1 leads to 4GlcNAc, Gal beta 1 leads to 4GlcNAc beta 1 leads to 6(Gal beta 1 leads to 4GlcNAc beta 1 leads to 2)Man alpha 1 leads to 6(Gal beta 1 leads to 4Glc NAc beta 1 leads to 2Man alpha 1 leads to 3)Man beta 1 leads to 4GlcNAc, Gal beta 1 leads to 4GlcNAc beta 1 leads to 6(Gal beta 1 leads to 4GlcNAc beta 1 leads to 2)Man alpha 1 leads to 6[Gal beta 1 leads to 4GlcNAc beta 1 leads to 4(Gal beta 1 leads to 4GlcNAc beta 1 leads to 2)Man alpha 1 leads to 3]Man beta 1 leads to 4GlcNAc, Gal beta 1 leads to 4GlcNAc beta 1 leads to 2Man alpha 1 leads to 6, and 3(Gal beta 1 leads to 4GlcNAc beta 1 leads to 3Gal beta 1 leads to 4GlcNAc beta 1 leads to 2Man alpha 1 leads to 3 and 6)Man beta 1 leads to 4GlcNAc, (formula see text) were elucidated. The amounts of total oligosaccharides excreted in the urine of the Type 2 subgroup were approximately one-tenth of those of Type 1. Moreover, the last eight oligosaccharides shown above, which have a Gal beta 1 leads to 4GlcNAc beta 1 leads to 3Gal beta 1 leads to 4GlcNAc beta 1 leads to outer chain, were completely missing in the urine of Type 2.

Topics: Carbohydrate Conformation; Carbohydrate Sequence; Chemical Phenomena; Chemistry; Child, Preschool; Female; G(M1) Ganglioside; Gangliosides; Gangliosidoses; Glycoside Hydrolases; Humans; Infant; Infant, Newborn; Male; Methods; Oligosaccharides

Data on the hydrolysis of fucosides, galactosides and arabinosides by different types of fucosidase are presented. The comparative study of the splitting of alpha-L-fucoside and beta-D-arabinoside with a similar structure at C1--C4 of the pyranose ring and the preparation of the enzyme by affinity chromatography showed that both substrates were hydrolysed by the same enzyme, alpha-L-fucoside(beta-D-arabinoside)hydrolase. The analogous investigation of the hydrolysis of beta-D-fucoside, beta-D-galactoside and alpha-L-arabinoside with a similar structure at C1--C4 of the pyranose ring demonstrated that these glycosides were split by the same enzyne, beta-D-fucoside(beta-D-galactoside, alpha-L-arabinoside)hydrolase, the activity of which is decreased dramatically in GM1-gangliosidosis. The data obtained support the assumption that the specific action of different types of fucosidase is due to recognition by these enzymes of C1--C4 of the pyranose ring in the corresponding substrates. The problems of differential diagnosis of some glycosidoses (focosidosis, GM1-gangliosidosis and Fabry disease) are discussed on the basis of the data obtained.

Topics: Adult; alpha-L-Fucosidase; Binding Sites; Child; Female; G(M1) Ganglioside; Galactosidases; Galactosides; Gangliosidoses; Glycoside Hydrolases; Humans; Kidney; Leukocytes; Male; Structure-Activity Relationship; Substrate Specificity

Among the seven oligosaccharide fractions obtained by Bio-Gel P-4 column chromatography of urine of GM1-gangliosidosis Type 1 patients, three fractions (peaks V, VI, and VII) were completely missing in the urine of GM1-gangliosidosis Type 2 patients. Structural study of these oligosaccharide fractions by sequential exoglycosidase digestion in combination with methylation analysis and periodate oxidation has shown that peaks V, VI, and VII are mixtures of 16, 30, and 49 isomeric oligosaccharides. All these 95 oligosaccharides contain Gal beta 1 leads to 4GlcNAc beta 1 lead to 3 repeating structures in their outer chain moieties, indicating that the tissues of GM1-gangliosidosis Type 2 patients do contain beta-galactosidase activity which releases readily galactose residue from such repeating sugar chains.

Topics: Carbohydrate Conformation; Carbohydrate Sequence; G(M1) Ganglioside; Gangliosides; Gangliosidoses; Glycoside Hydrolases; Humans; Methylation; Oligosaccharides

Clinical and pathological studies are reported from investigation of a 27-year-old man with GM1 gangliosidosis who experienced a slowly progressive dystonia that began about age 4, primarily affected the face and limbs, and eventually became almost totally incapacitating. There was only mild intellectual deterioration; myoclonus, seizures, and macular cherry-red spots were never observed. Postmortem examination revealed intraneuronal storage, localized predominantly to the basal ganglia, in which neurons contained round, multilamellated inclusions. Golgi studies revealed meganeurites arising from medium spiny neurons. Other areas of the central nervous system appeared relatively unaffected, although small basilar dilatations were observed in scattered cortical pyramidal neurons and Purkinje cell dendrites showed focal swellings. Vacuolated cells of the reticuloendothelial system were observed, including Kupffer cells and histiocytes in the spleen, marrow, and intestinal tract. Biochemical analysis revealed a generalized beta-galactosidase deficiency with specific accumulation of GM1 ganglioside in the basal ganglia.

Topics: Adult; Astrocytes; Brain; G(M1) Ganglioside; Gangliosides; Gangliosidoses; Humans; Kupffer Cells; Male; Microscopy, Electron; Neurons; Purkinje Cells

A patient with chronic GM1 gangliosidosis was studied enzymatically and biochemically. Leukocyte acid beta-galactosidase activity was severely deficient. In brain and liver, the 4-methylumbelliferyl beta-galactosidase with acidic pH optimum and lactosylceramidase II were deficient while other hydrolases were present in normal amounts, including sialidase determined with N-acetylneuramin-lactose and fetuin as substrates. Neutral beta-galactosidase in liver was increased up to fourfold over the control. Corresponding to the pathological findings, GM1 ganglioside sialic acid was increased in the basal ganglia to 57% of the total (normal, 12 to 16%), accounting for the rise in total ganglioside to 180% of normal in this origin. Only slight to moderate elevations in the proportion of GM1 ganglioside were noted in the cerebral cortex and white matter, without major increase in total ganglioside. Elevated asialo GM1 ganglioside was also confined to the basal ganglia. There was no increase in hepatic glycoproteins or in keratan sulfate-like materials. This is the only known patient with chronic GM1 gangliosidosis in whom abnormal accumulation of GM1 ganglioside has been demonstrated in affected tissue and sialidase deficiency has been excluded as the primary genetic defect.

Topics: Adult; beta-Galactosidase; Brain Chemistry; Chronic Disease; G(M1) Ganglioside; Gangliosides; Gangliosidoses; Humans; Lactose Intolerance; Leukocytes; Liver; Lysosomes; Male

Topics: alpha-L-Fucosidase; Arabinose; beta-Galactosidase; Clinical Enzyme Tests; G(M1) Ganglioside; Gangliosidoses; Glycoside Hydrolases; Humans; Lactose Intolerance; Leukocytes

Two cases of biochemically verified juvenile GM1-gangliosidosis are reported and the radiographic skeletal changes described in detail. Earlier reports are surveyed.

Topics: Bone and Bones; Child; Female; G(M1) Ganglioside; Gangliosidoses; Humans; Male; Radiography

Topics: Animals; Cats; Cerebral Cortex; Dendrites; Evoked Potentials; G(M1) Ganglioside; Gangliosidoses; Horseradish Peroxidase; Humans; Neural Inhibition; Neurons; Pyramidal Tracts; Thalamic Nuclei

Topics: G(M1) Ganglioside; Gangliosidoses; Humans; Mucopolysaccharidoses; Tay-Sachs Disease

Total ganglioside extracts prepared from brain tissue were concentrated either by dialysis against Carbowax or by employing Millipore filter cones. Thin-layer chromatography was then carried out using silica gel plates. After location of the various fractions quantitation was effected by direct densitometry. The methods that have been adopted are rapid and suitable for the study of brain gangliosides in post mortem and biopsy material in a clinical chemistry laboratory.

Topics: Animals; Brain; Cats; Chromatography, Thin Layer; G(M1) Ganglioside; G(M2) Ganglioside; Gangliosides; Gangliosidoses; Humans; Mucopolysaccharidosis III; Tay-Sachs Disease

Cultured skin fibroblasts from a 2-year-old boy with an atypical form of beta-galactosidase deficiency have been studied. With the artificial substrate 4-methylumbelliferyl-beta-D-galactopyranoside, 5--15% residual activity was found in fibroblasts from this patient. Most of this activity was in the monomeric A form of the enzyme, very little in the multimeric B form. Km value, pH profile, and heat lability of the mutant enzyme were similar to those of beta-galactosidase from control fibroblasts. Immunological studies showed that the mutant enzyme cross-reacted with an antiserum raised against human liver beta-galactosidase, but the catalytic activity per unit antigenic activity was lower than normal. It was demonstrated by somatic cell hybridization that the gene mutation in this patient is different from that in patients with type 1 or type 2 GM1-gangliosidosis. No genetic complementation was found after fusion of fibroblasts from this patient with those from two other clinical variants of GM1-gangliosidosis formerly designated type 3 and adult type 4.

Topics: Cells, Cultured; Child, Preschool; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Genetic Complementation Test; Genetic Variation; Humans; Kinetics; Male; Phenotype; Skin

Topics: Carbohydrates; Chromatography, Thin Layer; G(M1) Ganglioside; Gangliosidoses; Glycoproteins; Humans; Liver; Molecular Conformation; Oligosaccharides

Topics: Alleles; G(M1) Ganglioside; Gangliosides; Gangliosidoses; Heterozygote; Homozygote; Humans; Lactose Intolerance; Lysosomes; Models, Biological; Mutation; Phenotype

A clinical description of an apparently classical case of type 1 GM1 gangliosidosis is presented. The patient was the first-born child of first cousins. She was diagnosed at 6 weeks and died at 6 months. beta-Galactosidase activity was deficient in cultured fibroblasts using [3H]GM1 ganglioside and [3H]ceramide-lactose as substrates. Genetic complementation studies performed after cell fusion between cultured fibroblasts from the patient and from two other type 1, one type 2, and one juvenile GM1 gangliosidosis strain were positive with all strains. Subsequent studies revealed an increased excretion of a sialic acid-containing hexasaccharide in the patient's cells. Parents' fibroblasts contained normal levels of beta-galactosidase. The case emphasizes the variability of the clinical expression in sialidosis and the importance of demonstrating a primary gene defect in establishing a diagnosis of an inborn error or metabolism.

Topics: beta-Galactosidase; Consanguinity; Diagnosis, Differential; Female; Fibroblasts; G(M1) Ganglioside; Gangliosidoses; Genetic Complementation Test; Genotype; Humans; Infant, Newborn; Lactose Intolerance; Metabolism, Inborn Errors; Oligosaccharides; Phenotype; Sialic Acids

GM1-gangliosidosis is a disease characterized by abnormal accumulation of GM1-ganglioside in the brain and viscera. The disease is characterized by clinical findings similar to Hurler's disease and pathologic features resembling Niemann-Pick's disease but with involvement of the glomerular epithelium. A 14-month-old boy, clinically diagnosed as GM1-gangliosidosis, died of respiratory insufficiency and was autopsied except for the brain. Biochemically, marked increase of GM1-ganglioside in the viscera was demonstrated. Pathologically, the foam cells were present in the viscera. Some parts of the cytoplasmic vacuoles in the lungs and spleen contained osmiophilic fibrillar material electron-microscopically. This case was characterized by marked accumulation of foam cells in the pulmonary alveolar spaces.

Topics: Foam Cells; G(M1) Ganglioside; Gangliosides; Gangliosidoses; Humans; Infant; Lung; Lung Diseases; Male

Peculiar storage cells appearing in bone marrow aspirates from a patient with juvenile GM1-gangliosidosis and from one with beta-thalassemia were examined light microscopically, histochemically and electron microscopically. Light microscopically, most of the storage cells closely resembled Gaucher cells pathognomonic for Gaucher's disease. The cytoplasm of the Gaucher-like cells contained numerous variable-shaped membrane-bound inclusions mostly arranged in a mosaic pattern and filled with fibrillar materials. Intermingled tubular structures were usually narrow as compared to those of the Gaucher cells. These ultrastructural differences of the stored materials between the Gaucher-like cells and Gaucher cells were more clearly substantiated by the high resolution electron microscopy with negative staining technique. Enzyme cytochemically, acid phosphatase activity was proved in or around the storage inclusions, suggesting their lysosomal origin. Histochemically, it might be suggested that the stored materials of the Gaucher-like cells in juvenile GMI-gangliosidosis were non-sulfated acid mucopolysaccharides and glycopeptides, whereas glycoproteins were the major component of the storage cells in beta-thalassemia. Possible mechanisms of storage in the Gaucher-like cells were discussed in both disorders.

Topics: Child; Erythrocytes; Female; G(M1) Ganglioside; Gangliosides; Gangliosidoses; Gaucher Disease; Glycosaminoglycans; Histocytochemistry; Humans; In Vitro Techniques; Inclusion Bodies; Male; Periodic Acid-Schiff Reaction; Thalassemia

The activity of GM1 beta-galactosidase in the brain and liver of patients with GM1-gangliosidosis was assayed using GM1-ganglioside tritiated in the terminal galactose. In the cases of GM1-gangliosidosis Types 1 and 2A the activity was less than 0.5% of the control. In the liver of GM1-gangliosidosis Type 2B the activity was observed to be much higher than that of Types 1 and 2A. On Sephadex G-150 gel filtration, three active fractions (I, II and III) for 4-methylumbelliferyl beta-galactopyranoside (4MU) and two active fractions (I and II) for GM1-ganglioside were obtained in the control liver. There was no active fraction for GM1-ganglioside in spite of the preserved fraction I for 4MU in the liver of GM1-gangliosidosis Type 1 or Type 2A. In any of the three cases fraction II for both 4MU and G71-ganglioside was not detected.

Topics: Brain; Chromatography, Gel; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Humans; Hydrogen-Ion Concentration; Liver; Sodium Chloride; Taurocholic Acid

Topics: Enzyme Activation; G(M1) Ganglioside; Gangliosidoses; Genetic Variation; Hexosaminidases; Humans; Infant; Liver; Proteins; Substrate Specificity; Taurodeoxycholic Acid

Topics: Carbohydrates; Child; G(M1) Ganglioside; Gangliosides; Gangliosidoses; Humans; Male; Molecular Conformation; Oligosaccharides

Uptake of radioactivity from 14C-galactose into gangliosides by cultured skin fibroblasts was studied. GM3 was the major ganglioside in control human fibroblasts. An increase of GM1 was demonstrated in GM1-gangliosidosis fibroblasts. The degree of GM1 accumulation was correlated with the clinical types of this disease. The fibroblasts from an infantile-type patient showed a marked increase of GM1. In late-onset types the amount of total gangliosides was only slightly increased, but the distribution of individual gangliosides was definitely abnormal; a relative increase of GM1 was demonstrated in these cases. GM1 beta-galactosidase activities were not detectable in either infantile or late-onset cases.

Topics: beta-Galactosidase; Cells, Cultured; Child, Preschool; Female; Fibroblasts; G(M1) Ganglioside; G(M3) Ganglioside; Gangliosides; Gangliosidoses; Humans; Infant; Male; Skin

Topics: Animals; beta-Galactosidase; Biological Transport; Cats; Disease Models, Animal; Fibroblasts; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Humans; Liposomes; Lysosomes

Topics: Animals; Ataxia; Cerebral Cortex; Chromatography, Thin Layer; G(M1) Ganglioside; G(M2) Ganglioside; Gangliosides; Gangliosidoses; Heterozygote; Hexosaminidases; Humans; Leukocytes; Swine; Swine Diseases

Topics: beta-Galactosidase; Female; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Glycosaminoglycans; Humans; Infant

Electron microscope studies were carried out on neurons of the hippocampal formation in a feline mutant with beta-galactosidase deficiency and GMI-gangliosidosis. Fusiform processes with characteristics similar to meganeurites of Golgi studies were identified between cell bodies and axons of pyramidal and granule cells. The presence of dense material subjacent to the plasma membrane at the meganeurite-axon junction provides evidence that meganeurites form at the axon-hillock region and displace the initial axonal segment distally. Meganeurites of hippocampal neurons exhibited pleomorphic secondary processes with fine structural features of growth cones. Spines and spine-synapses were abundant on perikarya and meganeurites. Numerous membranous cytoplasmic bodies (MCBs) were encountered amongst otherwise normally appearing organelles of the cell body. MCBs were densely packed in meganeurites except near their peripheral area. They were less common in dendrites and rare in synapses of the neuropil. The observations provide further support for the view that meganeurites of mature cortical neurons in ganglioside storage diseases have embryonic growth characteristics.

Topics: Animals; Axons; Cat Diseases; Cats; Dendrites; Disease Models, Animal; Endoplasmic Reticulum; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Hippocampus; Humans; Neurons; Pyramidal Tracts; Synapses

Golgi studies were carried out on neurons in several forebrain structures of young adult mutant cats with inherited beta-galactosidase deficiency and neurobehavioral deterioration due to GM1-ganglioside storage disease. Meganeurites similar to those observed in several human gangliosidoses were present on small and medium pyramidal neurons, granule cells of the fascia dentata and spiny neurons of the caudate nucleus. Large and giant pyramidal cells of the motor cortex exhibited prominent somatic spines but lacked meganeurites. Cortical non-pyramidal neurons and aspiny caudate cells were relatively normal in appearance although they showed variable increases in cell body diameter. The range of morphological alterations in different types of cortical neurons in feline GM1-gangliosidosis was identical to that found in human ganglioside storage diseases. Neurite outgrowth from meganeurites was particularly prominent in the feline mutant. The extensive proliferation of neurites confined to meganeurites indicates that the latter have growth properties typical of embryonic neuronal elements. The demonstration of neurite outgrowth from meganeurites of mature cortical neurons in feline GM1-gangliosidosis suggests a possible role for gangliosides in neurite formation during neuronal differentiation and synaptogenesis.

Topics: Animals; Axons; Cat Diseases; Cats; Caudate Nucleus; Cerebral Cortex; Disease Models, Animal; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Hippocampus; Humans; Motor Cortex; Neurons; Pyramidal Tracts

Topics: Brain; Child, Preschool; Diagnosis, Differential; Female; G(M1) Ganglioside; Gangliosides; Gangliosidoses; Humans; Infant; Lactose Intolerance; Liver; Male

An assay procedure was developed for accurate estimation of lactosylceramidase II in the presence of relatively high activity of lactosylceramidase I. The procedure involves determination of lactosylceramide-cleaving activities under two different assay conditions, and lactosylceramidase II activity is calculated by the difference. Applicability of the procedure was evaluated with separated soluble fractions of the two beta-galactosidases from normal human brains, and with whole homogenates of gray and white matter, liver and cultured fibroblasts from control individuals and from patients with globoid cell leukodystrophy or GM1-gangliosidosis. The use of the lactosylceramidase I assay procedure developed by Wenger, D.A., Sattler, M., Clark, C. and McKelvey, H. ((1974) Clin. Chim. Acta 56, 199-206) and of the present procedure permits accurate diagnosis of both globoid cell leukodystrophy and GM1-gangliosidosis with one natural substrate, lactosylceramide, irrespective of the relative proportion of the two beta-galactosidases in the tissue.

Topics: beta-Galactosidase; Brain; Ceramides; Clinical Enzyme Tests; Fibroblasts; G(M1) Ganglioside; Galactosidases; Galactosylceramidase; Gangliosidoses; Humans; Isoenzymes; Kinetics; Lactose; Leukodystrophy, Globoid Cell; Liver

This report describes 7-month-old monozygotic twin female infants with GM1 gangliosidosis type I. In addition to the usual clinical and biochemical abnormalities generalized intracutaneous telangiectasis were present in both infants.

Topics: Diseases in Twins; Female; G(M1) Ganglioside; Galactosidases; Gangliosidoses; Humans; Infant; Phenotype; Pregnancy; Telangiectasis; Twins, Monozygotic

Topics: Animals; Cat Diseases; Cats; Cornea; Eye; G(M1) Ganglioside; Gangliosidoses; Humans; Retina

Topics: Animals; Brain; Cat Diseases; Cats; Cell Differentiation; G(M1) Ganglioside; Gangliosidoses; Humans; Neurons

GM1-ganglioside hydrolysis by leukocytes and fibroblasts, tissues easily obtainable from patients, was investigated using 3H-labeled GM1 and was found to be at least as active as that reported for any other tissue. Sodium taurocholate was required for the reaction, the crude bile salt at an optimum concentration of 0.4% producing twice as much activity as pure taurocholate at its optimum concentration of 0.8%. Leukocyte GM1-ganglioside beta-galactosidase and 4-MU-beta-gal cleaving activities were similar, 134.5 +/- 23.3 and 179.8 +/- 25.4 nmol/h/mg protein, respectively. In cultured skin fibroblasts and amniotic fluid cells these enzyme activities were 4 to 5 times higher. Homozygotes for GM1-gangliosidosis showed negligible activity while in heterozygotes the leukocyte GM1-cleaving activity was reduced to one-third of control values. In leukocytes from patients with four other sphingolipid storage diseases the activity was either normal (Krabbe's, Tay-Sachs, Metachromatic leukodystrophy) or increased (adult Gaucher's).

Topics: Adult; Animals; Cattle; G(M1) Ganglioside; Galactose; Galactosidases; Gangliosides; Gangliosidoses; Humans; Leukocytes; Skin; Sphingolipidoses; Taurocholic Acid

Topics: alpha-L-Fucosidase; Brain; Cells, Cultured; Fibroblasts; G(M1) Ganglioside; G(M2) Ganglioside; Gangliosidoses; Glycoproteins; Glycosphingolipids; Humans; Lipid Metabolism, Inborn Errors; Liver; Mannosidases

A 9-month-old dog with a history of progressive motor dysfunction was shown to have a deficiency in brain beta-galactosidase activity. The canine disease, like that of children with GM1 gangliosidosis, is characterized by accumulation of GM1 ganglioside in the brain, liver, and spleen, and membranous cytoplasmic bodies in neurons. The dog's pedigree suggests an autosomal recessive pattern of inheritance.

Topics: Animals; Brain; Brain Chemistry; Dog Diseases; Dogs; G(M1) Ganglioside; Galactosidases; Gangliosides; Gangliosidoses; Genes, Recessive; Humans; Liver; Pedigree; Spleen