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1.
Cell Stress Chaperones ; 23(5): 897-912, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29725981

RESUMO

The endoplasmic reticulum (ER) plays an important role in maintenance of proteostasis through the unfolded protein response (UPR), which is strongly activated in most neurodegenerative disorders. UPR signalling pathways mediated by IRE1α and ATF6 play a crucial role in the maintenance of ER homeostasis through the transactivation of an array of transcription factors. When activated, these transcription factors induce the expression of genes involved in protein folding and degradation with pro-survival effects. However, the specific contribution of these transcription factors to different neurodegenerative diseases remains poorly defined. Here, we characterised 44 target genes strongly influenced by XBP1 and ATF6 and quantified the expression of a subset of genes in the human post-mortem spinal cord from amyotrophic lateral sclerosis (ALS) cases and in the frontal and temporal cortex from frontotemporal lobar degeneration (FTLD) and Alzheimer's disease (AD) cases and controls. We found that IRE1α-XBP1 and ATF6 pathways were strongly activated both in ALS and AD. In ALS, XBP1 and ATF6 activation was confirmed by a substantial increase in the expression of both known and novel target genes involved particularly in co-chaperone activity and ER-associated degradation (ERAD) such as DNAJB9, SEL1L and OS9. In AD cases, a distinct pattern emerged, where targets involved in protein folding were more prominent, such as CANX, PDIA3 and PDIA6. These results reveal that both overlapping and disease-specific patterns of IRE1α-XBP1 and ATF6 target genes are activated in AD and ALS, which may be relevant to the development of new therapeutic strategies. Graphical abstract The endoplasmic reticulum (ER) plays an important role in maintenance of proteostasis through the unfolded protein response (UPR). Two major UPR signalling pathways are mediated by IRE1α and ATF6. Here, we demonstrate that these pathways activate differential gene sets in human post-mortem tissues derived from amyotrophic lateral sclerosis (ALS) compared to Alzheimer's disease (AD) cases. Our results identify IRE1α and ATF6 specific targets that can have major implications in the development of new therapeutic strategies and potential biomarkers.


Assuntos
Doença de Alzheimer/genética , Esclerose Lateral Amiotrófica/genética , Estresse do Retículo Endoplasmático/genética , Resposta a Proteínas não Dobradas/genética , Fator 6 Ativador da Transcrição/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Doença de Alzheimer/metabolismo , Esclerose Lateral Amiotrófica/metabolismo , Endorribonucleases/metabolismo , Lobo Frontal/metabolismo , Degeneração Lobar Frontotemporal/genética , Degeneração Lobar Frontotemporal/metabolismo , Humanos , Pessoa de Meia-Idade , Proteínas Serina-Treonina Quinases/metabolismo , Elementos de Resposta , Medula Espinal/metabolismo , Lobo Temporal/metabolismo , Ativação Transcricional , Proteína 1 de Ligação a X-Box/metabolismo , Adulto Jovem
2.
Psychiatry Res ; 246: 335-340, 2016 Dec 30.
Artigo em Inglês | MEDLINE | ID: mdl-27750116

RESUMO

Previously, we found a significant gender-specific association of schizophrenia, in a UK case/control study, with SLC30A3, a candidate that is consistently down-regulated in schizophrenia in two independent cohorts. In view of the potential significance of this finding, we extended this study to a larger cohort using GWAS data from the Psychiatric Genetic Consortium (PGC). Meta-analysis was performed for the only two SLC30A3 SNP variants (rs11126936 and rs11126929) available in most PGC cohorts. A significant association with schizophrenia was found for both variants. When meta-analysis was performed in male and female case-control subsets, an increased and gender-specific effect of allele on risk of disease was found in females for both SNPs with no significant effect in males, which was further associated with a gender-specific effect on gene expression. In conclusion, using a large European-wide sample we were able to replicate the gender-specific association previously found in a UK cohort.


Assuntos
Proteínas de Transporte de Cátions/genética , Estudo de Associação Genômica Ampla , Esquizofrenia/genética , Estudos de Casos e Controles , Europa (Continente) , Feminino , Humanos , Masculino , Fatores Sexuais
3.
Eur Psychiatry ; 29(3): 172-8, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23849395

RESUMO

Previous microarray analysis of gene expression in frontal cortex showed differential expression of genes associated with synaptic function in schizophrenia compared to matched-controls in two independent cohorts. One of these genes validated in both cohorts, SLC30A3, which encodes the Zinc Transporter 3 (ZNT3), is localised to synaptic vesicles in glutamate synapses and known to be involved in cognitive function. In view of the robust depletion of SLC30A3 mRNA in two independent studies and the importance of this gene in cognitive function, we investigated whether single nucleotide polymorphism (SNP) associations with schizophrenia could be detected in a UK case controlled schizophrenia cohort. Four SNPs were selected across this gene and genotyped in a cohort of cases and controls from East UK. We found significant associations with schizophrenia at the allelic (ORs: 1.51 to 1.57), genotype (ORs: 1.46 to 1.53) and haplotype level (P=2.15×10(-4)). These associations proved to be gender-specific with significant effects of allele (ORs: 1.74 to 2.11), genotype (ORs: 1.78 to 2.14) and haplotype (P=3.51×10(-5)) observed in female schizophrenia cases but not males, when split by gender. In conclusion, SNPs in SLC30A3 showed a gender-specific association with schizophrenia in this East UK cohort, which merits further investigation in other population samples.


Assuntos
Proteínas de Transporte de Cátions/genética , Esquizofrenia/genética , Alelos , Estudos de Casos e Controles , Estudos de Coortes , Inglaterra/epidemiologia , Feminino , Estudo de Associação Genômica Ampla , Genótipo , Haplótipos/genética , Humanos , Masculino , Polimorfismo de Nucleotídeo Único/genética , Esquizofrenia/epidemiologia , Fatores Sexuais
4.
Mol Psychiatry ; 14(12): 1083-94, 2009 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-19255580

RESUMO

Schizophrenia is a severe psychiatric disorder with a world-wide prevalence of 1%. The pathophysiology of the illness is not understood, but is thought to have a strong genetic component with some environmental influences on aetiology. To gain further insight into disease mechanism, we used microarray technology to determine the expression of over 30 000 mRNA transcripts in post-mortem tissue from a brain region associated with the pathophysiology of the disease (Brodmann area 10: anterior prefrontal cortex) in 28 schizophrenic and 23 control patients. We then compared our study (Charing Cross Hospital prospective collection) with that of an independent prefrontal cortex dataset from the Harvard Brain Bank. We report the first direct comparison between two independent studies. A total of 51 gene expression changes have been identified that are common between the schizophrenia cohorts, and 49 show the same direction of disease-associated regulation. In particular, changes were observed in gene sets associated with synaptic vesicle recycling, transmitter release and cytoskeletal dynamics. This strongly suggests multiple, small but synergistic changes in gene expression that affect nerve terminal function.


Assuntos
Regulação da Expressão Gênica/genética , Terminações Nervosas/metabolismo , Córtex Pré-Frontal/patologia , Esquizofrenia/genética , Esquizofrenia/patologia , Sinapses/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Coortes , Biologia Computacional/métodos , Intervalos de Confiança , Feminino , Perfilação da Expressão Gênica/métodos , Humanos , Masculino , Pessoa de Meia-Idade , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Córtex Pré-Frontal/metabolismo , RNA Mensageiro/metabolismo , Sinapses/metabolismo
5.
Neurology ; 66(6): 839-44, 2006 Mar 28.
Artigo em Inglês | MEDLINE | ID: mdl-16421333

RESUMO

OBJECTIVE: To perform genetic linkage analysis in a family affected with ALS and frontotemporal dementia (FTD). METHODS: The authors performed a genome-wide linkage analysis of a four-generation, 50-member Scandinavian family in which five individuals were diagnosed with ALS and nine with FTD. Linkage calculations assuming autosomal dominant inheritance of a single neurodegenerative disease manifesting as either ALS or FTD with age-dependent penetrance were performed. Further analyses for ALS alone and FTD alone were performed. A parametric logarithm of odds (lod) score of 2.0 or greater was required for further study of a potential locus and crossover (haplotype) analysis. RESULTS: A new ALS-FTD locus was identified between markers D9s1870 and D9s1791 on human chromosome 9p21.3-p13.3. A maximum multipoint lod score of 3.00 was obtained between markers D9s1121 and D9s2154. Crossover analysis indicates this region covers approximately 21.8 cM, or 14Mb. CONCLUSIONS: A locus on chromosome 9p21.3-p13.3 is linked to ALS-FTD.


Assuntos
Esclerose Lateral Amiotrófica/genética , Cromossomos Humanos Par 9/genética , Demência/genética , Predisposição Genética para Doença/genética , Locos de Características Quantitativas/genética , Idoso , Feminino , Ligação Genética/genética , Marcadores Genéticos/genética , Haplótipos/genética , Humanos , Masculino , Pessoa de Meia-Idade , Linhagem
6.
Brain Res Bull ; 56(3-4): 299-306, 2001.
Artigo em Inglês | MEDLINE | ID: mdl-11719264

RESUMO

Expansion of unstable DNA regions containing trinucleotide/tandem repeats (TNRs) represents a common genetic mutation in hereditary forms of neurological disorders. The spectrum of neurological diseases linked to TNR expansions has recently broadened to include conditions with both dominant and recessive inheritance and those with or without clinical anticipation. In view of the frequent involvement of the spinal cord in neurodegenerative disorders, we have analysed this key tissue to identify pathological TNRs. We have used two approaches to isolate a wide range of trinucleotide/tandem repeat-containing transcripts (TNRTs) from human spinal cord, firstly a polymerase chain reaction (PCR)-based method and secondly by screening a spinal cord cDNA library immobilised on a membrane. Overall, 97 TNRTs belonging to a number of key protein families, the most highly represented being transcription factors, intracellular signalling molecules and cytoskeletal proteins, have been isolated most of which have not previously been considered as potential disease-causing genes. The commonest repeat motifs found in our study were CAG (37%) and CCG (24%). Known genes involved in DNA repeat expansion-related neurological disorders (e.g., AAD10, Ataxin-3, Huntingtin) were detected which validated our methods. We have characterised homogeneous TNRs among the detected gene candidates in a search for potential pathological repeat expansions. The potential role of the gene candidates identified is discussed in terms of their contribution to neurodegenerative processes.


Assuntos
Doenças Neurodegenerativas/genética , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Medula Espinal/fisiologia , Repetições de Trinucleotídeos , Biblioteca Gênica , Testes Genéticos , Humanos , Proteínas do Tecido Nervoso/genética , Análise de Sequência com Séries de Oligonucleotídeos/normas , Peptídeos/genética , Reação em Cadeia da Polimerase , Reprodutibilidade dos Testes
7.
Brain Res Mol Brain Res ; 93(2): 148-63, 2001 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-11589992

RESUMO

Kainate-induced status epilepticus is associated with both apoptotic and necrotic cell death and induction of heat shock proteins (HSPs) in hippocampal and cortical regions of the rodent brain. In the present study we have examined the temporal, spatial and cellular expression patterns of mRNAs for the highly inducible HSPs, HSP70 and HSP27, together with the apoptotic marker, caspase 3 (CPP32) in rat brain after systemic administration of kainate. HSP70 mRNA was transiently induced in the forebrain by kainate, principally in the CA1, CA3 and hilar cells of the hippocampal formation, in piriform cortex and discrete thalamic nuclei. Maximal expression was seen at 8 h after kainate which then declined to background levels by 7 days. Labelling was predominantly neuronal. In contrast, HSP27 mRNA expression was more widespread. Intense labelling was observed in CA1, CA3 and the hilar region at 8 h after kainate but the expression profile for HSP27 mRNA expanded considerably with intense signals seen in corpus callosum, cortex and thalamus at 24 h post kainate. Emulsion autoradiographs indicated a predominantly glial localisation for HSP27 mRNA. In the hilus, a distinct subpopulation of interneurones were found to express HSP27 mRNA. CPP32 mRNA was upregulated in CA1, CA3 and hilus of the hippocampal formation and in piriform cortex. CPP32 mRNA expression was more restricted and similar in distribution to HSP70 mRNA being localised to neurones. The present study demonstrates the unique early expression of HSP27 mRNA by glial cells and distinct populations of neurones which extends beyond those in which HSP70 and CPP32 induction occurs with subsequent cell loss.


Assuntos
Química Encefálica/efeitos dos fármacos , Caspases/biossíntese , Agonistas de Aminoácidos Excitatórios/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Proteínas de Choque Térmico HSP70/biossíntese , Proteínas de Choque Térmico/biossíntese , Ácido Caínico/farmacologia , Proteínas do Tecido Nervoso/biossíntese , Neuroglia/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Animais , Autorradiografia/métodos , Caspase 3 , Caspases/genética , Córtex Cerebral/citologia , Córtex Cerebral/efeitos dos fármacos , Córtex Cerebral/metabolismo , Agonistas de Aminoácidos Excitatórios/toxicidade , Proteína Glial Fibrilar Ácida/biossíntese , Proteína Glial Fibrilar Ácida/genética , Ácido Glutâmico/fisiologia , Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico/genética , Hipocampo/citologia , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Hibridização In Situ , Injeções Intraperitoneais , Ácido Caínico/toxicidade , Masculino , Proteínas do Tecido Nervoso/genética , Neuroglia/metabolismo , Neurônios/metabolismo , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley , Convulsões/induzido quimicamente , Tálamo/citologia , Tálamo/efeitos dos fármacos , Tálamo/metabolismo , Fatores de Tempo
8.
J Neurochem ; 77(1): 132-45, 2001 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-11279269

RESUMO

In order to obtain insight into the aetiology and pathogenesis of amyotrophic lateral sclerosis (ALS), high-density gene discovery arrays (GDA human version 1.2) containing 18 400 non-redundant EST cDNAs pooled from different tissue libraries have been used to monitor gene expression in lumbar spinal cord from ALS cases compared with controls. Quantitative filter analysis revealed differential expression of cDNAs normalized to internal standards. These candidates have been further investigated and their expression in spinal cord characterized in a panel of ALS and control subjects. Significant differential expression was obtained for 14 genes, 13 being elevated (up to six-fold) and one decreased (by 80%) in ALS. Amongst those elevated in ALS were thioredoxin and glial fibrilary acid protein, which have already been shown to be up-regulated in ALS, thus supporting the reliability of this approach. The other differentially regulated transcripts confirmed in the expression studies represent potential candidates in ALS pathogenesis being involved in antioxidant systems, neuroinflammation, the regulation of motor neurone function, lipid metabolism, protease inhibition and protection against apoptosis. The use of the GDA system has greatly facilitated the screening and retrieval of sequence information and has generated useful information on the cascade of molecular events occurring in ALS and potentially may highlight new candidates playing a role in the aetiology and progression of this disease.


Assuntos
Esclerose Lateral Amiotrófica/genética , Esclerose Lateral Amiotrófica/metabolismo , Perfilação da Expressão Gênica , Peptídeos e Proteínas de Sinalização Intercelular , Análise de Sequência com Séries de Oligonucleotídeos , Medula Espinal/metabolismo , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Esclerose Lateral Amiotrófica/patologia , Regulação para Baixo , Etiquetas de Sequências Expressas , Feminino , Expressão Gênica , Proteína Glial Fibrilar Ácida/genética , Proteína Glial Fibrilar Ácida/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Hibridização de Ácido Nucleico/métodos , Proteína Plasmática A Associada à Gravidez/genética , Progranulinas , Proteína-Lisina 6-Oxidase/genética , RNA Mensageiro/biossíntese , Medula Espinal/patologia , Tiorredoxinas/genética , Tiorredoxinas/metabolismo , Tubulina (Proteína)/genética , Regulação para Cima , Proteínas Virais/genética , Proteínas Virais/metabolismo
9.
J Neurochem ; 75(6): 2511-20, 2000 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-11080204

RESUMO

We have recently isolated a 2.2-kb cDNA clone (1C5) from a human spinal cord cDNA library with partial identity to the 14-3-3 protein mRNA encoding the theta protein (YWHAQ). 14-3-3 protein transcripts are highly expressed in large projection neurones of the hippocampus, cerebellum, and spinal cord and have been found to be significantly up-regulated in rat motor neurones following hypoglossal nerve axotomy. In this study we investigated whether the 1C5 transcript (YWHAQ) isolated from spinal cord was involved in amyotrophic lateral sclerosis (ALS). We found a significant up-regulation of 1C5 (YWHAQ) in lumbar spinal cord from patients with sporadic ALS compared with controls, with the highest levels of expression being found in individuals with predominant lower motor neurone involvement. A 6-bp tandem repeat in the 5'-untranslated region of the gene was found to be polymorphic, but no significant association with disease was found following genomic analysis of this region. The localisation of 1C5 (YWHAQ) to chromosome 2 was determined and coincides with that reported for clone HS1 (EMBL accession no. X57347). These results show the marked up-regulation of the 14-3-3 isoform (YWHAQ) in ALS spinal cord and indicate the involvement of a potential 14-3-3-mediated survival pathway in the pathogenesis of ALS.


Assuntos
Esclerose Lateral Amiotrófica/genética , RNA Mensageiro/metabolismo , Medula Espinal/metabolismo , Tirosina 3-Mono-Oxigenase/metabolismo , Proteínas 14-3-3 , Adolescente , Idoso , Idoso de 80 Anos ou mais , Alelos , Sequência de Aminoácidos , Esclerose Lateral Amiotrófica/metabolismo , Sequência de Bases , Feminino , Lobo Frontal/química , Lobo Frontal/metabolismo , Lobo Frontal/patologia , Perfilação da Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Especificidade de Órgãos/genética , Mapeamento Físico do Cromossomo , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Análise de Sequência de DNA , Medula Espinal/química , Sequências de Repetição em Tandem/genética , Repetições de Trinucleotídeos/genética , Tirosina 3-Mono-Oxigenase/genética
10.
Eur J Hum Genet ; 8(8): 649-51, 2000 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-10951529

RESUMO

Refsum's disease (MIM 266500) is a recessive disorder characterised by defective peroxisomal alpha-oxidation of phytanic acid. A Refsum's disease gene, phytanoyl-CoA hydroxylase (PAHX), has been localised to chromosome 10p13 between the markers D10S226-D10S223. This study investigated whether all cases of Refsum's disease were linked with chromosome 10p13. Eight genetically informative families comprising 92 individuals including 17 living patients with a Refsum's disease phenotype and initial plasma phytanic acid > 200 micromol/L were recruited. Linkage to the 10pter-10p11.2 region was investigated using a panel of eight dinucleotide repeat markers. Linkage analysis of this phenotypically identical cohort suggested that Refsum's disease was genetically heterogeneous (Zmax = 5.28, alpha = 0.45). Two subgroups were identified. One group of four families with eight affected individuals had a maximum multipoint lod score for linkage of 3.89 in the region D10S547 to D10S191, whilst in another three families with nine affected individuals linkage to this region was definitely excluded. Our results show that Refsum's disease is genetically heterogeneous, with up to 55% of cases not being linked to the PAHX gene locus at D10S547 to D10S223. This suggests that Refsum's disease, in common with other peroxisomal 'diseases', may be more accurately described as a heterogeneous syndrome.


Assuntos
Cromossomos Humanos Par 10/genética , Heterogeneidade Genética , Oxigenases de Função Mista/genética , Doença de Refsum/genética , Mapeamento Cromossômico , Consanguinidade , Feminino , Genes Recessivos , Ligação Genética , Humanos , Escore Lod , Masculino , Repetições de Microssatélites , Linhagem , Doença de Refsum/enzimologia
11.
Neuropathol Appl Neurobiol ; 26(3): 258-72, 2000 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-10886684

RESUMO

The vulnerability of motoneurones to glutamate has been implicated in neurological disorders such as amyotrophic lateral sclerosis but it is not known whether specific receptor subtypes mediate this effect. In order to investigate this further, the expression of N-methyl-D-aspartate (NMDA) receptor subunits was studied during the first three post-natal weeks when motoneurones are differentially vulnerable to injury following neonatal nerve crush compared to the adult. Unilateral nerve crush was carried out at day 2 after birth (P2) which causes a decrease of 66% in motoneurone number by 14 days (P14). To study receptor expression in identified motoneurones, serial section analysis was carried out on retrogradely labelled common peroneal (CP) motoneurones by combined immunocytochemistry and in situ hybridization (ISH). mRNA levels were also quantified in homogenates from lumbar spinal cords in which the side ipsilateral to the crush was separated from the contralateral side. The NR1 subunit of the NMDA receptor was widely distributed in the spinal cord being expressed most strongly in motoneurone somata particularly during the neonatal period (P3-P7). The NR2 subunits were also expressed at higher levels in the somata and dendrites of neonatal motoneurones compared to older animals. NR2B mRNA was expressed at low to moderate levels throughout the studied period whereas NR2A mRNA levels were low until P21. Following unilateral nerve crush, an initial decrease in NR1 mRNA occurred at one day after nerve crush (P3) in labelled CP motoneurones ipsilateral to the crush which was followed by a significant increase in NR1 subunit expression at 5 days post-injury. This increase was bilateral although reaching greater significance ipsilateral to the crush compared with sham-operated animals. A significant increase in NR1 and NR2B mRNA post injury was also detected in spinal cord homogenates. In addition, the changes in levels of NR1 and NR2B mRNA were reflected by comparable bilateral changes at P7 in receptor protein determined by quantitative immunocytochemical analysis of NR1 and NR2 subunit expression in identified CP motoneurones indicating a co-ordinated regulation of receptor subunits in response to injury.


Assuntos
Neurônios Motores/metabolismo , Nervo Fibular/lesões , Receptores de N-Metil-D-Aspartato/genética , Medula Espinal/metabolismo , Fatores Etários , Animais , Animais Recém-Nascidos , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Hibridização In Situ , Músculo Esquelético/inervação , Compressão Nervosa , RNA Mensageiro/análise , Ratos , Ratos Sprague-Dawley , Medula Espinal/citologia , Traumatismos da Medula Espinal/metabolismo , Traumatismos da Medula Espinal/patologia
12.
DNA Cell Biol ; 19(5): 265-73, 2000 May.
Artigo em Inglês | MEDLINE | ID: mdl-10855793

RESUMO

In order to identify trinucleotide- and tandem repeat-containing transcripts in human spinal cord, hybridization of a high-density spinal cord cDNA library filter was carried out using a radioactively labeled degenerate oligonucleotide designed to detect different trinucleotide repeats including those known to occur in disease-associated expansions, in a single step. The sequence analysis of the trinucleotide repeat-containing transcripts (TNRTs) revealed 23 known mammalian genes with trinucleotide repeat-containing regions (TNRs), some of which were not previously reported to contain TNRs, and 18 cDNA clones with no or insignificant sequence homology to known genes. Amongst the known genes detected was the fragile X gene (FMR-1) containing (CGG)30. Other genes containing extended TNRs of 9 to 21 repeats were calcium-dependent protease, ATBF1-A, ferritin H chain, and the G protein Gsalpha2. Ten sequences containing perfect TNRs and two sequences containing perfect tandem repeats (derived from 11 TNRTs) were further analyzed for allelic variation using primers flanking the TNR, and five were shown to exhibit two to five alleles per TNR. These transcripts were further investigated for their chromosomal localization where unknown or only partially characterized. The transcripts that were polymorphic in the TNR region were ATBF1-A (a homeodomain protein), clone 390013 on chromosome Xp11, a member of the family of the 14.3.3 protein kinase C regulators, a human translation initiation factor (an isolog of the yeast Suilisol gene 1), and a novel sequence (TR21). Only the first two transcripts showed the presence of rare expanded alleles. Characterization of polymorphic TNRs in novel and even known genes expressed in human spinal cord is likely to help in the identification of new candidates for genes involved in neurodegenerative disorders.


Assuntos
Medula Espinal/fisiologia , Sequências de Repetição em Tandem/genética , Repetições de Trinucleotídeos/genética , Mapeamento Cromossômico , Cromossomos Humanos Par 2 , DNA Complementar/análise , Biblioteca Gênica , Proteínas de Homeodomínio/genética , Humanos , Hibridização de Ácido Nucleico , Fases de Leitura Aberta , Polimorfismo Genético , Proteína Quinase C , RNA Mensageiro/genética , Cromossomo X
13.
Neuroreport ; 10(16): 3265-8, 1999 Nov 08.
Artigo em Inglês | MEDLINE | ID: mdl-10599831

RESUMO

In the light of earlier findings of reduced cholecystokinin (CCK) peptide and CCK mRNA levels in the cerebral cortex, we have used in situ hybridization to examine possible regulation of mRNAs coding for two isoforms of the CCK(B) receptor in frontal cortex (Brodmann's area 10) of schizophrenic patients. The hybridizations revealed a 51% decrease of the full length CCK(B) receptor mRNA in the outer layers (II-III) of the frontal cortex. The corresponding alterations for the truncated isoform were a 65% reduction in the outer layers and a 62% reduction in the inner layers (IV-VI) of the frontal cortex. This strengthens the hypothesis that CCKergic transmission in this part of the brain is involved in the pathophysiology of schizophrenia.


Assuntos
Química Encefálica/fisiologia , RNA Mensageiro/biossíntese , Receptores da Colecistocinina/biossíntese , Esquizofrenia/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Autorradiografia , Feminino , Lobo Frontal/metabolismo , Humanos , Hibridização In Situ , Isomerismo , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Receptor de Colecistocinina B , Inclusão do Tecido
14.
Eur J Hum Genet ; 7(1): 12-9, 1999 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-10094186

RESUMO

Expansions of trinucleotide CAG repeats have been demonstrated in at least eight neurodegenerative disorders, and suggested to occur in several others, including bipolar disorder and schizophrenia. Chromosome 18 loci have been implicated in bipolar disorder pedigrees by linkage analysis. To address this putative link between chromosome 18 CAG trinucleotide repeats and neuropsychiatric illness, we have screened a chromosome 18 cosmid library (LL18NCO2" AD") and identified 14 novel candidate loci. Characterisation of these loci involved repeat flank sequencing, estimation of polymorphism frequency and mapping using FISH as well as radiation hybrid panels. These mapped trinucleotide loci will be useful in the investigation of chromosome 18 in neurodegenerative or psychiatric conditions, and will serve to integrate physical and radiation hybrid maps of chromosome 18.


Assuntos
Mapeamento Cromossômico , Cromossomos Humanos Par 18 , Repetições de Trinucleotídeos , Sequência de Bases , Primers do DNA , Humanos , Células Híbridas , Transtornos Mentais/genética , Homologia de Sequência do Ácido Nucleico , Sitios de Sequências Rotuladas
15.
J Biol Chem ; 274(8): 5061-9, 1999 Feb 19.
Artigo em Inglês | MEDLINE | ID: mdl-9988753

RESUMO

Overexpression of the gene encoding the 70-kDa heat shock protein (hsp70) has previously been shown to protect neuronal cells against subsequent thermal or ischemic stress. It has no protective effect, however, against stimuli that induce apoptosis, although a mild heat shock (sufficient to induce hsp synthesis) does have a protective effect against apoptosis. We have prepared disabled herpes simplex virus-based vectors that are able to produce high level expression of individual hsps in infected neuronal cells without damaging effects. We have used these vectors to show that hsp27 and hsp56 (which have never previously been overexpressed in neuronal cells) as well as hsp70 can protect dorsal root ganglion neurons from thermal or ischemic stress. In contrast, only hsp27 can protect dorsal root ganglion neurons from apoptosis induced by nerve growth factor withdrawal, and hsp27 also protects the ND7 neuronal cell line from retinoic acid-induced apoptosis. However, hsp70 showed no protective effect against apoptosis in contrast to its anti-apoptotic effect in non-neuronal cell types. These results thus identify hsp27 as a novel neuroprotective factor and show that it can mediate this effect when delivered via a high efficiency viral vector.


Assuntos
Apoptose/genética , Vetores Genéticos , Proteínas de Choque Térmico/genética , Neurônios/citologia , Simplexvirus/genética , Animais , Linhagem Celular , Cricetinae , Ratos , Ratos Sprague-Dawley
16.
J Neural Transm (Vienna) ; 105(6-7): 689-701, 1998.
Artigo em Inglês | MEDLINE | ID: mdl-9826111

RESUMO

In order to identify possible morphological changes which occur in the anterior horn of normal individuals during aging, 40 controls with no neurological disease were studied. Brain and spinal cord tissue was processed according to a brain banking protocol. Controls were grouped according to age in 10 year intervals. Serial sections (20 microm) of formalin fixed, paraffin-embedded tissue were obtained, from each cervical, thoracic and lumbar spinal cord segment. Every 5th section (until 2 mm) was stained with haematoxylin and eosin and the numbers of motor neurons in the anterior horn counted at x400 magnification. Descriptive statistical analysis was performed using the SPSS program. Parallel sections (5 microm) of the same spinal segments were immunostained with a panel of antibodies including glial fibrillary acidic protein (GFAP), tau, ubiquitin and two phosphorylated neurofilaments subunits. Significant neuronal loss with aging was found by regression line analysis where three equations were used to calculate the number of motor neurons by age in each spinal segment. In 24/40 cases spheroids were observed and they were more numerous in the lumbar segment. GFAP staining revealed a distinctive cellular pattern in the anterior horn of oldest cases. Large and intensely stained astrocytes were seen in the anterior horn of cases aged over 75 years. The number of astrocytes increased progressively with age up to 70 years. Some of the changes observed in the present study may be the result of a selective vulnerability of large motor neurons to aging which could play an important role in the progression of MND. Most of these changes may also have similar pathophysiological mechanisms.


Assuntos
Envelhecimento/fisiologia , Astrócitos/citologia , Citoesqueleto/ultraestrutura , Neurônios Motores/ultraestrutura , Medula Espinal/citologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Contagem de Células , Feminino , Proteína Glial Fibrilar Ácida/metabolismo , Humanos , Técnicas Imunológicas , Masculino , Pessoa de Meia-Idade , Neurônios Motores/citologia , Proteínas de Neurofilamentos/metabolismo , Neurônios/citologia , Fosforilação , Medula Espinal/metabolismo , Medula Espinal/ultraestrutura , Ubiquitinas/metabolismo , Proteínas tau/metabolismo
17.
Eur J Neurosci ; 10(11): 3343-50, 1998 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-9824447

RESUMO

The heat shock proteins (HSPs) are induced by stressful stimuli and have a protective effect. Different HSPs protect with different efficiencies against different stresses indicating that optimal protection would be obtained with a non-stressful agent which induced a range of HSPs. We have prepared a herpesvirus vector expressing a constitutively active mutant form of heat shock factor 1 (HSF1) which, unlike the wild-type form of this transcription factor, does not require stress for its activation. Upon infection of neuronal cells, this virus induced a more restricted range of HSPs than in non-neuronal cells. Infection with the virus protected neuronal cells against subsequent thermal or ischaemic stress in accordance with its ability to induce HSP70 expression but did not protect them against apoptotic stimuli. The mechanisms of these effects and their significance for the use of HSF to manipulate HSP gene expression is discussed.


Assuntos
Apoptose/fisiologia , Proteínas de Ligação a DNA/biossíntese , Proteínas de Choque Térmico/biossíntese , Temperatura Alta/efeitos adversos , Isquemia/patologia , Neurônios/patologia , Estresse Fisiológico/patologia , Animais , Animais Recém-Nascidos , Western Blotting , Linhagem Celular , Proteínas de Ligação a DNA/genética , Vetores Genéticos , Fatores de Transcrição de Choque Térmico , Técnicas In Vitro , Camundongos , Mutação , Neurônios/metabolismo , Neurônios/virologia , Ratos , Ratos Sprague-Dawley , Simplexvirus/genética , Raízes Nervosas Espinhais/patologia , Fatores de Transcrição
18.
Neuropathol Appl Neurobiol ; 24(5): 353-8, 1998 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-9821165

RESUMO

A patient presented with features of olivopontocerebellar atrophy and was found to have marked hyperglycinaemia. Severe atrophy of the cerebellum and brain stem was found at post-mortem, with numerous glial cytoplasmic inclusions (GCIs) in atrophic areas, characteristic of multiple system atrophy. In situ hybridization studies of the spinal cord demonstrated a selective reduction in expression of glycine transporter mRNA. We suggest that the resulting impairment of regulation of glycine concentrations at synaptic level resulted in excitotoxic damage to neurons.


Assuntos
Sistemas de Transporte de Aminoácidos Neutros , Proteínas de Transporte/genética , Glicina/sangue , Atrofias Olivopontocerebelares/metabolismo , Atrofias Olivopontocerebelares/patologia , Proteínas de Transporte/metabolismo , Ventrículos Cerebrais/patologia , Feminino , Expressão Gênica/fisiologia , Proteínas da Membrana Plasmática de Transporte de Glicina , Humanos , Hibridização In Situ , Pessoa de Meia-Idade , Neurotransmissores/genética , Neurotransmissores/metabolismo , RNA Mensageiro/análise , Medula Espinal/química , Medula Espinal/metabolismo , Medula Espinal/patologia , Substância Negra/patologia
20.
Neuropharmacology ; 37(6): 739-44, 1998 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-9707287

RESUMO

The inducible form of cyclo-oxygenase (COX-2) mRNA is rapidly induced in the spinal cord following peripheral inflammation produced by intraplantar injection of Freund's complete adjuvant (FCA). COX-2 mRNA induction is also accompanied by increased prostaglandin (PG) levels which are closely correlated with behavioural indicators of increased pain sensitivity. The aim of this study was to determine whether the anti-inflammatory glucocorticoid, dexamethasone, which acts locally to prevent the development of oedema would also reduce the associated central changes characterised by the induction of COX-2 mRNA and PGs. Unilateral intraplantar FCA induced a marked oedema evident from 2 h to 7 days after FCA injection which was significantly attenuated by dexamethasone pretreatment at all time points. Dexamethasone also significantly prevented the induction of COX-2 mRNA (2 4 h) and elevated levels of prostaglandins (6-keto PGF1alpha) in lumbar spinal cord (8 h). In this study we have confirmed the anti-inflammatory effect of dexamethasone and linked this to central changes in gene expression relevant to the development of altered pain thresholds following intraplantar FCA.


Assuntos
Anti-Inflamatórios/farmacologia , Dexametasona/farmacologia , Glucocorticoides/farmacologia , Isoenzimas/biossíntese , Prostaglandina-Endoperóxido Sintases/biossíntese , Prostaglandinas/biossíntese , Medula Espinal/efeitos dos fármacos , Animais , Anti-Inflamatórios/uso terapêutico , Ciclo-Oxigenase 2 , Dexametasona/uso terapêutico , Edema/induzido quimicamente , Edema/prevenção & controle , Adjuvante de Freund , Glucocorticoides/uso terapêutico , Injeções Espinhais , Masculino , RNA Mensageiro/metabolismo , Ratos , Medula Espinal/metabolismo , Doenças da Medula Espinal/induzido quimicamente , Doenças da Medula Espinal/prevenção & controle
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