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1.
Nat Commun ; 5: 4824, 2014 Sep 12.
Article in English | MEDLINE | ID: mdl-25215604

ABSTRACT

Proteinaceous inclusions are common hallmarks of many neurodegenerative diseases. TDP-43 proteinopathies, consisting of several neurodegenerative diseases, including frontotemporal lobar dementia (FTLD) and amyotrophic lateral sclerosis (ALS), are characterized by inclusion bodies formed by polyubiquitinated and hyperphosphorylated full-length and truncated TDP-43. The structural properties of TDP-43 aggregates and their relationship to pathogenesis are still ambiguous. Here we demonstrate that the recombinant full-length human TDP-43 forms structurally stable, spherical oligomers that share common epitopes with an anti-amyloid oligomer-specific antibody. The TDP-43 oligomers are stable, have exposed hydrophobic surfaces, exhibit reduced DNA binding capability and are neurotoxic in vitro and in vivo. Moreover, TDP-43 oligomers are capable of cross-seeding Alzheimer's amyloid-ß to form amyloid oligomers, demonstrating interconvertibility between the amyloid species. Such oligomers are present in the forebrain of transgenic TDP-43 mice and FTLD-TDP patients. Our results suggest that aside from filamentous aggregates, TDP-43 oligomers may play a role in TDP-43 pathogenesis.


Subject(s)
Amyloid/chemistry , Cerebral Cortex/pathology , DNA-Binding Proteins/chemistry , Frontotemporal Dementia/pathology , Protein Aggregation, Pathological/pathology , TDP-43 Proteinopathies/pathology , Amino Acid Sequence , Amyloid/immunology , Animals , Antibodies, Monoclonal/biosynthesis , Antibodies, Monoclonal/chemistry , Cell Line, Tumor , Cerebral Cortex/chemistry , Cerebral Cortex/immunology , DNA-Binding Proteins/genetics , DNA-Binding Proteins/immunology , Embryo, Mammalian , Epitopes/chemistry , Epitopes/immunology , Escherichia coli/genetics , Escherichia coli/metabolism , Frontotemporal Dementia/genetics , Frontotemporal Dementia/immunology , Gene Expression , HEK293 Cells , Humans , Injections, Intraventricular , Male , Mice , Molecular Sequence Data , Neurons/chemistry , Neurons/immunology , Neurons/pathology , Primary Cell Culture , Protein Aggregates , Protein Aggregation, Pathological/genetics , Protein Aggregation, Pathological/immunology , Recombinant Fusion Proteins/chemistry , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/immunology , TDP-43 Proteinopathies/genetics , TDP-43 Proteinopathies/immunology
2.
Hum Mol Genet ; 23(23): 6366-74, 2014 Dec 01.
Article in English | MEDLINE | ID: mdl-25008110

ABSTRACT

During postnatal development, neuronal activity controls the remodeling of initially imprecise neuronal connections through the regulation of gene expression. MeCP2 binds to methylated DNA and modulates gene expression during neuronal development and MECP2 mutation causes the autistic disorder Rett syndrome. To investigate a role for MeCP2 in neuronal circuit refinement and to identify activity-dependent MeCP2 transcription regulations, we leveraged the precise organization and accessibility of olfactory sensory axons to manipulation of neuronal activity through odorant exposure in vivo. We demonstrate that olfactory sensory axons failed to develop complete convergence when Mecp2 is deficient in olfactory sensory neurons (OSNs) in an otherwise wild-type animal. Furthermore, we demonstrate that expression of selected adhesion genes was elevated in Mecp2-deficient glomeruli, while acute odor stimulation in control mice resulted in significantly reduced MeCP2 binding to these gene loci, correlating with increased expression. Thus, MeCP2 is required for both circuitry refinement and activity-dependent transcriptional responses in OSNs.


Subject(s)
Methyl-CpG-Binding Protein 2/metabolism , Olfactory Bulb/metabolism , Sensory Receptor Cells/metabolism , Animals , Axons/metabolism , Axons/ultrastructure , Cadherins/metabolism , Methyl-CpG-Binding Protein 2/genetics , Mice , Mice, Transgenic , Nerve Tissue Proteins/metabolism , Odorants , Olfactory Bulb/cytology , Protocadherins , Sensory Receptor Cells/ultrastructure , Transcription, Genetic
3.
Alcohol Clin Exp Res ; 38(6): 1540-9, 2014 Jun.
Article in English | MEDLINE | ID: mdl-24730561

ABSTRACT

BACKGROUND: Alcoholic steatohepatitis (ASH) is caused in part by the effects of ethanol (EtOH) on hepatic methionine metabolism. METHODS: To investigate the phenotypic and epigenetic consequences of altered methionine metabolism in this disease, we studied the effects of 4-week intragastric EtOH feeding with and without the methyl donor betaine in cystathionine beta synthase (CßS) heterozygous C57BL/6J mice. RESULTS: The histopathology of early ASH was induced by EtOH feeding and prevented by betaine supplementation, while EtOH feeding reduced and betaine supplementation maintained the hepatic methylation ratio of the universal methyl donor S-adenosylmethionine (SAM) to the methyltransferase inhibitor S-adenosylhomocysteine (SAH). MethylC-seq genomic sequencing of heterozygous liver samples from each diet group found 2 to 4% reduced methylation in gene bodies, but not promoter regions of all autosomes of EtOH-fed mice, each of which were normalized in samples from mice fed the betaine-supplemented diet. The transcript levels of nitric oxide synthase (Nos2) and DNA methyltransferase 1 (Dnmt1) were increased, while those of peroxisome proliferator receptor-α (Pparα) were reduced in EtOH-fed mice, and each was normalized in mice fed the betaine-supplemented diet. DNA pyrosequencing of CßS heterozygous samples found reduced methylation in a gene body of Nos2 by EtOH feeding that was restored by betaine supplementation and was correlated inversely with its expression and positively with SAM/SAH ratios. CONCLUSIONS: The present study has demonstrated relationships among EtOH induction of ASH with aberrant methionine metabolism that was associated with gene body DNA hypomethylation in all autosomes and was prevented by betaine supplementation. The data imply that EtOH-induced changes in selected gene transcript levels and hypomethylation in gene bodies during the induction of ASH are a result of altered methionine metabolism that can be reversed through dietary supplementation of methyl donors.


Subject(s)
Betaine/therapeutic use , DNA Methylation/drug effects , Ethanol/pharmacology , Fatty Liver, Alcoholic/metabolism , Gene Expression Regulation/drug effects , Homocystinuria/metabolism , Animals , DNA (Cytosine-5-)-Methyltransferase 1 , DNA (Cytosine-5-)-Methyltransferases/analysis , Dietary Supplements , Liver/chemistry , Liver/drug effects , Methionine/metabolism , Mice , Mice, Inbred C57BL , Nitric Oxide Synthase Type II/analysis , PPAR alpha/analysis , S-Adenosylhomocysteine/metabolism , S-Adenosylmethionine/metabolism
4.
Hepatology ; 57(2): 555-65, 2013 Feb.
Article in English | MEDLINE | ID: mdl-22945834

ABSTRACT

UNLABELLED: Hepatic methionine metabolism may play an essential role in regulating methylation status and liver injury in Wilson's disease (WD) through the inhibition of S-adenosylhomocysteine hydrolase (SAHH) by copper (Cu) and the consequent accumulation of S-adenosylhomocysteine (SAH). We studied the transcript levels of selected genes related to liver injury, levels of SAHH, SAH, DNA methyltransferases genes (Dnmt1, Dnmt3a, Dnmt3b), and global DNA methylation in the tx-j mouse (tx-j), an animal model of WD. Findings were compared to those in control C3H mice, and in response to Cu chelation by penicillamine (PCA) and dietary supplementation of the methyl donor betaine to modulate inflammatory and methylation status. Transcript levels of selected genes related to endoplasmic reticulum stress, lipid synthesis, and fatty acid oxidation were down-regulated at baseline in tx-j mice, further down-regulated in response to PCA, and showed little to no response to betaine. Hepatic Sahh transcript and protein levels were reduced in tx-j mice with consequent increase of SAH levels. Hepatic Cu accumulation was associated with inflammation, as indicated by histopathology and elevated serum alanine aminotransferase (ALT) and liver tumor necrosis factor alpha (Tnf-α) levels. Dnmt3b was down-regulated in tx-j mice together with global DNA hypomethylation. PCA treatment of tx-j mice reduced Tnf-α and ALT levels, betaine treatment increased S-adenosylmethionine and up-regulated Dnmt3b levels, and both treatments restored global DNA methylation levels. CONCLUSION: Reduced hepatic Sahh expression was associated with increased liver SAH levels in the tx-j model of WD, with consequent global DNA hypomethylation. Increased global DNA methylation was achieved by reducing inflammation by Cu chelation or by providing methyl groups. We propose that increased SAH levels and inflammation affect widespread epigenetic regulation of gene expression in WD.


Subject(s)
DNA Methylation/drug effects , Liver/metabolism , Methionine/metabolism , Adenosylhomocysteinase/antagonists & inhibitors , Adenosylhomocysteinase/metabolism , Animals , Betaine/metabolism , Betaine/pharmacology , Copper/metabolism , Copper/pharmacology , DNA (Cytosine-5-)-Methyltransferases/metabolism , Disease Models, Animal , Down-Regulation , Endoplasmic Reticulum Stress , Epigenesis, Genetic/drug effects , Hepatolenticular Degeneration/metabolism , Hepatolenticular Degeneration/pathology , Inflammation/metabolism , Mice , Mice, Inbred C3H , Penicillamine/pharmacology , S-Adenosylhomocysteine/metabolism , DNA Methyltransferase 3B
5.
Environ Mol Mutagen ; 53(8): 589-98, 2012 Oct.
Article in English | MEDLINE | ID: mdl-22930557

ABSTRACT

Persistent organic pollutants (POPs), including polychlorinated biphenyls (PCBs) and polybrominated diphenylethers (PBDEs) that bioaccumulate in lipid-rich tissues are of concern as developmental neurotoxicants. Epigenetic mechanisms such as DNA methylation act at the interface of genetic and environmental factors implicated in autism-spectrum disorders. The relationship between POP levels and DNA methylation patterns in individuals with and without neurodevelopmental disorders has not been previously investigated. In this study, a total of 107 human frozen postmortem brain samples were analyzed for eight PCBs and seven PBDEs by GC-micro electron capture detector and GC/MS using negative chemical ionization. Human brain samples were grouped as neurotypical controls (n = 43), neurodevelopmental disorders with known genetic basis (n = 32, including Down, Rett, Prader-Willi, Angelman, and 15q11-q13 duplication syndromes), and autism of unknown etiology (n = 32). Unexpectedly, PCB 95 was significantly higher in the genetic neurodevelopmental group, but not idiopathic autism, as compared to neurotypical controls. Interestingly, samples with detectable PCB 95 levels were almost exclusively those with maternal 15q11-q13 duplication (Dup15q) or deletion in Prader-Willi syndrome. When sorted by birth year, Dup15q samples represented five out of six of genetic neurodevelopmental samples born after the 1976 PCB ban exhibiting detectable PCB 95 levels. Dup15q was the strongest predictor of PCB 95 exposure over age, gender, or year of birth. Dup15q brain showed lower levels of repetitive DNA methylation measured by LINE-1 pyrosequencing, but methylation levels were confounded by year of birth. These results demonstrate a novel paradigm by which specific POPs may predispose to genetic copy number variation of 15q11-q13.


Subject(s)
Brain/drug effects , Brain/metabolism , Child Development Disorders, Pervasive/genetics , Chromosomes, Human, Pair 15/genetics , Halogenated Diphenyl Ethers/toxicity , Polychlorinated Biphenyls/toxicity , Adolescent , Adult , Child , Child, Preschool , DNA Copy Number Variations/drug effects , DNA Copy Number Variations/genetics , DNA Methylation/drug effects , DNA Methylation/genetics , Epigenesis, Genetic/drug effects , Epigenesis, Genetic/genetics , Female , Humans , Male , Middle Aged , Prader-Willi Syndrome/genetics , Young Adult
6.
Hum Mol Genet ; 21(11): 2399-411, 2012 Jun 01.
Article in English | MEDLINE | ID: mdl-22343140

ABSTRACT

The widespread use of persistent organic polybrominated diphenyl ethers (PBDEs) as commercial flame retardants has raised concern about potential long-lived effects on human health. Epigenetic mechanisms, such as DNA methylation, are responsive to environmental influences and have long-lasting consequences. Autism spectrum disorders (ASDs) have complex neurodevelopmental origins whereby both genetic and environmental factors are implicated. Rett syndrome is an X-linked ASD caused by mutations in the epigenetic factor methyl-CpG binding protein 2 (MECP2). In this study, an Mecp2 truncation mutant mouse (Mecp2(308)) with social behavioral defects was used to explore the long-lasting effects of PBDE exposure in a genetically and epigenetically susceptible model. Mecp2(308/+) dams were perinatally exposed daily to 2,2',4,4'-tetrabromodiphenyl ether 47 (BDE-47) and bred to wild-type C57BL/6J males, and the offspring of each sex and genotype were examined for developmental, behavioral and epigenetic outcomes. Perinatal BDE-47 exposure negatively impacted fertility of Mecp2(308/+) dams and preweaning weights of females. Global hypomethylation of adult brain DNA was observed specifically in female offspring perinatally exposed to BDE-47 and it coincided with reduced sociability in a genotype-independent manner. A reversing interaction of Mecp2 genotype on BDE-47 exposure was observed in a short-term memory test of social novelty that corresponded to increased Dnmt3a levels specifically in BDE-47-exposed Mecp2(308/+) offspring. In contrast, learning and long-term memory in the Morris water maze was impaired by BDE-47 exposure in female Mecp2(308/+) offspring. These results demonstrate that a genetic and environmental interaction relevant to social and cognitive behaviors shows sexual dimorphism, epigenetic dysregulation, compensatory molecular mechanisms and specific behavioral deficits.


Subject(s)
Epigenomics , Methyl-CpG-Binding Protein 2/genetics , Mutation , Polybrominated Biphenyls/toxicity , Animals , Animals, Newborn , Behavior, Animal , Brain/drug effects , Brain/metabolism , DNA (Cytosine-5-)-Methyltransferases/genetics , DNA (Cytosine-5-)-Methyltransferases/metabolism , DNA Methyltransferase 3A , Environmental Pollutants/toxicity , Female , Halogenated Diphenyl Ethers , Male , Maternal Exposure/adverse effects , Maze Learning , Methyl-CpG-Binding Protein 2/metabolism , Mice , Mice, Inbred C57BL , Mice, Inbred Strains , Polybrominated Biphenyls/adverse effects
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