galactocerebroside and Disease-Models--Animal

Remyelination is the regenerative process by which demyelinated axons are reinvested with new myelin sheaths. It is associated with functional recovery and maintenance of axonal health. It occurs as a spontaneous regenerative response following demyelination in a range of pathologies including traumatic injury as well as primary demyelinating disease such as multiple sclerosis (MS). Experimental models of demyelination based on the use of toxins, while not attempting to accurately mimic a disease with complex etiology and pathogenesis such as MS, have nevertheless proven extremely useful for studying the biology of remyelination. In this chapter, we review the main toxin models of demyelination, drawing attention to their differences and how they can be used to study different aspects of remyelination. We also describe the optimal use of these models, highlighting potential pitfalls in interpretation, and how remyelination can be unequivocally recognized. Finally, we discuss the role of toxin models alongside viral and immune-mediated models of demyelination.

Topics: Animals; Antibodies; Cats; Demyelinating Diseases; Disease Models, Animal; Ethidium; Galactosylceramides; Humans; Lysophosphatidylcholines; Mice; Mice, Inbred C57BL; Myelin Sheath; Nerve Regeneration; Rabbits; Rats; Spinal Cord

Meta-analysis studies showed that smokers have increased risk for developing Alzheimer's disease (AD) compared with non-smokers, and neuroimaging studies revealed that smoking damages white matter structural integrity.. The present study characterizes the effects of side-stream (second hand) cigarette smoke (CS) exposures on the expression of genes that regulate oligodendrocyte myelin-synthesis, maturation, and maintenance and neuroglial functions.. Adult male A/J mice were exposed to air (8 weeks; A8), CS (4 or 8 weeks; CS4, CS8), or CS8 followed by 2 weeks recovery (CS8 + R). The frontal lobes were used for histology and qRT-PCR analysis.. Luxol fast blue, Hematoxylin and Eosin stained histological sections revealed CS-associated reductions in myelin staining intensity and narrowing of the corpus callosum. CS exposures broadly decreased mRNA levels of immature and mature oligodendrocyte myelin-associated, neuroglial, and oligodendrocyte-related transcription factors. These effects were more prominent in the CS8 compared with CS4 group, suggesting that molecular abnormalities linked to white matter atrophy and myelin loss worsen with duration of CS exposure. Recovery normalized or upregulated less than 25% of the suppressed genes; in most cases, inhibition of gene expression was either sustained or exacerbated.. CS exposures broadly inhibit expression of genes needed for myelin synthesis and maintenance. These adverse effects often were not reversed by short-term CS withdrawal. The results support the hypothesis that smoking contributes to white matter degeneration, and therefore could be a key risk factor for a number of neurodegenerative diseases, including AD.

Topics: 2',3'-Cyclic-Nucleotide Phosphodiesterases; AC133 Antigen; Animals; Antigens, CD; Brain; Disease Models, Animal; Galactosylceramides; Gene Expression Regulation; Glycoproteins; Leukoencephalopathies; Male; Mice; Nerve Degeneration; Nerve Tissue Proteins; Neuroglia; Neurons; Nicotiana; Oligodendroglia; Peptides; Smoking; Transcription Factors

Krabbe's disease (KD) is a degenerative lysosomal storage disease resulting from deficiency of β-galactocerebrosidase activity. Over 100 mutations are known to cause the disease, and these usually occur in compound heterozygote patterns. In affected patients, nonsense mutations leading to a nonfunctional enzyme are often found associated with other mutations. The twitcher mouse is a naturally occurring model of KD, containing in β-galactocerebrosidase a premature stop codon, W339X. Recent studies have shown that selected compounds may induce the ribosomal bypass of premature stop codons without affecting the normal termination codons. The rescue of β-galactocerebrosidase activity induced by treatment with premature termination codon (PTC) 124, a well-characterized compound known to induce ribosomal read-through, was investigated on oligodendrocytes prepared from twitcher mice and on human fibroblasts from patients bearing nonsense mutations. The effectiveness of the nonsense-mediated mRNA decay (NMD) inhibitor 1 (NMDI1), a newly identified inhibitor of NMD, was also tested. Incubation of these cell lines with PTC124 and NMDI1 increased the levels of mRNA and rescued galactocerebrosidase enzymatic activity in a dose-dependent manner. The low but sustained expression of β-galactocerebrosidase in oligodendrocytes was sufficient to improve the morphology of the differentiated cells. Our in vitro approach provides the basis for further investigation of ribosomal read-through as an alternative therapeutic strategy to ameliorate the quality of life in selected KD patients. © 2016 Wiley Periodicals, Inc.

Topics: Animals; Animals, Newborn; Cell Line, Transformed; Codon, Terminator; Disease Models, Animal; Dose-Response Relationship, Drug; Fibroblasts; Galactosylceramidase; Galactosylceramides; Humans; Leukodystrophy, Globoid Cell; Mice; Mice, Inbred C57BL; Mice, Mutant Strains; Nonsense Mediated mRNA Decay; Oligodendroglia; Oxadiazoles; RNA, Messenger

Metachromatic leukodystrophy (MLD) is a lysosomal storage disease caused by genetic deficiency of arylsulfatase A (ARSA) enzyme. Failure in catalyzing the degradation of its major substrate, sulfatide (Sulf), in oligodendrocytes and Schwann cells leads to severe demyelination in the peripheral (PNS) and central nervous system (CNS), and early death of MLD patients. The ARSA knockout mice develop a disease that resembles MLD but is milder, without significant demyelination in the PNS and CNS. We showed that adeno-associated virus serotype 5-mediated gene transfer in the brain of ARSA knockout mice reverses Sulf storage and prevents neuropathological abnormalities and neuromotor disabilities when vector injections are performed at a pre-symptomatic stage of disease. Direct injection of viral particles into the brain of ARSA knockout mice at a symptomatic stage results in sustained expression of ARSA, prevention of Sulf storage and neuropathological abnormalities. Despite these significant corrections, the treated mice continue to develop neuromotor disability. We show that more subtle biochemical abnormalities involving gangliosides and galactocerebroside are in fact not corrected.

Topics: Animals; Brain; Brain Chemistry; Cerebroside-Sulfatase; Dependovirus; Disease Models, Animal; Galactosylceramides; Gangliosides; Genetic Therapy; Genetic Vectors; Immunohistochemistry; Injections; Leukodystrophy, Metachromatic; Mice; Mice, Knockout; Mice, Mutant Strains; Motor Activity; Transduction, Genetic; Treatment Failure

To probe the effects of possible inhibitors or enhancers of in vivo myelination, we have modified a technique widely used in studies of the developing neuromuscular system that involves incorporation of test compounds into a silicon rubber solution, which solidifies on contact with air. U-shaped rubber implants are inserted around the sciatic nerve of 1-day-old rats and left in place for 24-48 h. Sections from the region of the nerve lying within the implant, with or without the test compound, are then immunolabeled, examined with in situ hybridization or electron microscopy. Application of EDTA (440 microg/implant) in this way strongly suppressed the levels of the myelin-associated molecules protein P0, myelin basic protein (MBP), and galactocerebroside (Galc). mRNA levels for P0 and the myelin-related transcription factor Krox-20 were also reduced, further supporting association of the EDTA-induced effect with the myelinating Schwann cells. In contrast, no obvious differences were observed in either neurofilament (NF) protein or glial fibrillary acidic protein (GFAP) expression, suggesting absence of influence on axons or nonmyelinating Schwann cells. Despite the severely altered molecular composition of myelin in the presence of EDTA, examination in the electron microscope did not reveal any apparent ultrastructural changes in the myelin sheaths or nerve development. This work introduces a novel method for studying nerve development and shows that EDTA, which chelates divalent cations such as Ca(2+) and Mg(2+), strongly and selectively reduces levels of molecules, which, on postnatal days 1-4, are expressed in myelinating cells at much higher levels than in cells not engaged in myelination.

Topics: Animals; Animals, Newborn; Chelating Agents; Disease Models, Animal; DNA-Binding Proteins; Down-Regulation; Drug Carriers; Early Growth Response Protein 2; Edetic Acid; Galactosylceramides; Growth Inhibitors; Implants, Experimental; Microscopy, Electron, Transmission; Myelin Basic Protein; Myelin P0 Protein; Myelin Proteins; Myelin Sheath; Nerve Regeneration; Neurosurgical Procedures; Rats; RNA, Messenger; Schwann Cells; Sciatic Nerve; Silicone Elastomers; Transcription Factors

Neurological deficit in experimental allergic encephalomyelitis (EAE) and multiple sclerosis (MS) is probably a consequence of synergy between T and B cell responses to CNS antigens. During the demyelinating phase of chronic relapsing EAE in ABH mice, anti-myelin oligodendrocyte glycoprotein (MOG) responses were increased compared to the inflammatory acute phase, but such levels did not correlate with the severity of clinical disease. The pathogenicity of antibodies (Ab) to MOG, myelin basic protein (MBP), proteolipid protein (PLP) and galactocerebroside (GalC) was investigated in vivo following injection at the onset of EAE. An IgG2a monoclonal Ab (mAb), clone Z12, directed to MOG augmented clinical disease and demyelination in ABH and C57BL/6 mice, but not MOG knock-out mice. No effect was observed with F(ab(2))' fragments of Z12 or with the anti-MOG IgG1 mAbs, clones Y10 or 8-18C5. Cobra venom factor partially reduced the augmenting effect of mAb Z12 suggesting a role for complement. The pathogenic effect of anti-myelin Abs was not restricted to MOG since an anti-GalC mAb exacerbated inflammation in the CNS while an MBP mAb (clone 22) reduced clinical disease. Taken together, these data provide further evidence that myelin-reactive Abs generated during autoimmune neurological disease may play an important role not only in the pathogenesis of disease but also the regulation of myelin-targeted autoimmune disease.

Topics: Animals; Autoantibodies; Complement System Proteins; Demyelinating Diseases; Disease Models, Animal; Elapid Venoms; Encephalomyelitis, Autoimmune, Experimental; Female; Galactosylceramides; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Myelin Basic Protein; Myelin Proteins; Myelin Proteolipid Protein; Myelin-Associated Glycoprotein; Myelin-Oligodendrocyte Glycoprotein; Recurrence; Spinal Cord

Eight sooty mangabey monkeys were inoculated intravenously and intradermally with varying doses of Mycobacterium leprae from 4.8 x 10(7) to 4.8 x 10(10). Serum samples were obtained from the animals at intervals of about 3 months for 90 months, and were examined for IgM and IgG antibodies to nerve antigens, including ceramide, galactocerebroside (GC), and asialo-GM1 (AGM1), using an enzyme-linked immunosorbent assay (ELISA). The serological results were then compared with clinical findings, particularly nerve involvement. Of 8 mangabey monkeys inoculated with M. leprae, 7 animals had clinical leprosy; 6 of them had nerve damage, including neurologic deformities in 4 monkeys and nerve enlargement in 2. Median time for the initial signs of leprosy was 10 months postinoculation (p.i.), a range from 4 to 35 months. In contrast, nerve damage was noted rather late, about 35 to 86 months p.i. (median 54 months). The major immunoglobulin class to ceramide, GC, and AGM1 antigens was IgM, and the antibody responses to the nerve antigens appeared from 15 to 63 months p.i. (median 37 months). Antineural antibodies were thus detectable about 18 months (range -2 to 60 months) prior to observable nerve damage. In addition, elevation of antineural antibody levels were predictive of clinical exacerbation of the disease and neuritic damage. This study suggests that antineural antibodies are produced during the course of M. leprae infection and may be indicative of nerve damage, such as neurological deformities or nerve enlargement, in leprosy patients.

Topics: Animals; Autoantigens; Brain Diseases; Ceramides; Cercocebus atys; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; G(M1) Ganglioside; Galactosylceramides; Immunoglobulin G; Immunoglobulin M; Leprosy, Lepromatous; Mycobacterium leprae; Nerve Tissue Proteins

Experimental demyelination was induced by intraneural injection of anti-galactocerebroside serum into the sciatic nerves of rats. Schwann cells undergoing mitotic division were observed between days 3 to 9 after the injection and demyelinated segments were still associated with macrophages. Dividing Schwann cells were often present in association with both unmyelinated and myelinated fibers. Whether or not, daughter Schwann cells migrate along the same fiber towards neighboring demyelinated segments remains unclear. When Schwann cells attached to axon membranes of demyelinated segments were studied at later time points, they were present in clusters randomly at various regions of the segments. There was no proximo-distal gradient for the wave of Schwann cell proliferation. Mean Schwann cell internuclear distances were around 40-50 microns at the earliest time of remyelination. Schwann cell redistribution and remyelination progressed regardless of the length of demyelinated segments.

Topics: Animals; Demyelinating Diseases; Disease Models, Animal; Galactosylceramides; Immune Sera; Male; Microscopy, Electron; Mitosis; Peripheral Nervous System Diseases; Rats; Schwann Cells

Enzyme replacement with liposomes containing beta-galactosidase obtained from charonia lumpas was carried out in murine globoid cell leukodystrophy (GLD). Charonia lumpas beta-galactosidase was able to hydrolyze galactocerebroside trapped into liposomes prepared from lecithin, cholesterol and sulfatide (molar ratio; 7:2:1). Liposomes containing charonia lumpas beta-galactosidase were successfully incorporated into the mouse tissues. 3H-galactocerebroside labeled liposomes were also incorporated into mouse liver, spleen and other tissues. The accumulation rate of 3H-galactocerebroside into twithcer mice liver and spleen was almost 40 to 100 times higher than those of controls and degraded to 70 to 80% of accumulated radioactivity of 3H-galactocerebroside by single injection of liposomes containing charonia lumpas beta-galactosidase. Results suggest that exogeneous enzyme trapped in liposomes can be useful for the correction of accumulated compound.

Topics: Animals; Aspergillus oryzae; beta-Galactosidase; Brain; Cattle; Cerebrosides; Disease Models, Animal; Fabaceae; Galactosidases; Galactosylceramides; Hydrolysis; Kidney; Leukodystrophy, Globoid Cell; Liposomes; Liver; Mice; Mice, Neurologic Mutants; Mollusca; Plants, Medicinal; Spleen

A model of immune-mediated optic nerve demyelination is described. Micro-injection of small volumes (less than 5 microliter) of high titer polyclonal anti-Gal-C serum into the cat optic nerve resulted in a focal, highly selective demyelinative lesion followed by remyelination. Demyelination appears to be due to a dual effect on myelin and on oligodendrocytes. The numbers of these cells within the lesion were initially reduced but subsequently increased as remyelination occurred.

Topics: Animals; Cats; Cerebrosides; Demyelinating Diseases; Disease Models, Animal; Galactosylceramides; Immune Sera; Microscopy, Electron; Optic Nerve; Optic Nerve Diseases