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1.
Bratisl Lek Listy ; 121(2): 107-110, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32115961

RESUMO

AIM: Diabetes is one of the most common diseases which can attenuate brain function by destroying hippocampus neurons, while reelin is a largely secreted extracellular matrix glycoprotein in the hippocampus causing synaptic plasticity, promoting postsynaptic structures and maturing neurons. The aim of this study was to assess the effect of exercise, as an external factor for neurogenesis in the brain, on reelin levels and memory improvement in diabetic rats. METHOD: Thirty rats were randomly allocated into three groups; healthy sedentary, diabetic sedentary and diabetic exercise-trained. The experimental group was treadmill-exercised at speed 22 m/min for 1 hour, 5 days per week. Finally, spatial memory of rats was tested and reelin levels were measured. RESULTS: The results showed that short-term exercise improved spatial memory in diabetic rats but had no effect on reelin levels in the hippocampus of diabetic rats. CONCLUSION: Diabetes reduced the spatial memory without altering the reelin levels while exercise improved spatial memory without altering the reelin levels (Fig. 4, Ref. 33).


Assuntos
Moléculas de Adesão Celular Neuronais/metabolismo , Diabetes Mellitus Experimental , Proteínas da Matriz Extracelular/metabolismo , Hipocampo , Proteínas do Tecido Nervoso/metabolismo , Condicionamento Físico Animal , Serina Endopeptidases/metabolismo , Memória Espacial , Animais , Hipocampo/metabolismo , Neurogênese , Plasticidade Neuronal , Ratos
2.
Life Sci ; 249: 117518, 2020 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-32147432

RESUMO

AIMS: The objectives of the present study were to investigate the mechanisms of Ninj-1 regulation in TNFα-activated human endothelial cells (HEC), and to test if Amlodipine (AML) ameliorates the inflammatory stress by decreasing Ninj-1 expression. MAIN METHODS: TNFα-activated HEC with/without AML (0.1 µM and 1 µM) were used. TNFα-receptor 1 (TNFR1) was silenced and inhibitors for oxidative stress (N-acetyl cysteine), endoplasmic reticulum stress (salubrinal, 4-phenyl butyric acid), or NF-kB (Bay 11-7085) and p38 MAPK (SB203580) were used. Levels of Ninj-1, TNFR1, monocyte adhesion, endoplasmic reticulum stress (ERS) sensors, NADPH oxidase- and mitochondria-derived oxidative species were evaluated. KEY FINDINGS: The novel findings that we report here are: (i) silencing the endothelial TNFR1 leads to decreased Ninj-1 expression and diminished monocyte adhesion; (ii) increased oxidative stress, ERS and NF-kB activation enhance Ninj-1 expression and monocyte adhesion; (iii) up-regulation of endothelial Ninj-1 expression stimulates monocytes adhesion to TNFα - activated HEC; (iv) AML diminishes monocyte adhesion by reducing Ninj-1 expression through mechanisms involving the decrease of NADPH oxidase and mitochondria-dependent oxidative stress, ERS and NF-kB. In addition, AML alleviates apoptosis by reducing the pro-apoptotic CHOP expression and re-establishing the mitochondrial transmembrane potential. SIGNIFICANCE: The results of the present study suggest that Ninj-1 and the proteins involved in its regulation can be considered therapeutic targets for the alleviation of inflammation- dependent disorders. In addition, we demonstrate that some of the benefic effects of AML can be achieved through regulation of Ninj-1.


Assuntos
Anlodipino/farmacologia , Moléculas de Adesão Celular Neuronais/fisiologia , Adesão Celular/fisiologia , Monócitos/citologia , Fatores de Crescimento Neural/fisiologia , Receptores Tipo I de Fatores de Necrose Tumoral/fisiologia , Fator de Necrose Tumoral alfa/fisiologia , Regulação para Cima , Vasodilatadores/farmacologia , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Inativação Gênica , Células Endoteliais da Veia Umbilical Humana , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , NADPH Oxidases/metabolismo , NF-kappa B/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Receptores Tipo I de Fatores de Necrose Tumoral/genética
3.
Ann Otol Rhinol Laryngol ; 129(7): 653-656, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32028786

RESUMO

OBJECTIVES: Congenital midline cervical cleft (CMCC) is a rare congenital anterior neck anatomical anomaly. We present the case of two related patients (grandchild and maternal grandmother) who were both born with a congenital midline cervical cleft along with genetic analysis. METHODS: Clinical examination of both patients and surgical excision of the grandchild was performed. Genetic analysis with exome sequencing (ES) was conducted for both patients. RESULTS: Genetic analysis with exome sequencing (ES) revealed apparently novel single nucleotide variants in 66 genes present in both proband and grandmother. Five of these variants are predicted to cause frameshifting in the coding region of the respective genes and truncated proteins (OVGP1, TYW1B, ZAN, SSPO, FOLR3). Two of these genes (TYW1B and SSPO) have homozygous indel mutations in both patients. CONCLUSIONS: To our knowledge, this is the first case of two related patients with a congenital midline cervical cleft. The results of our genetic analysis reveal potential relevance to CMCC development.


Assuntos
Região Branquial/anormalidades , Proteínas de Transporte/genética , Moléculas de Adesão Celular Neuronais/genética , Anormalidades Craniofaciais/genética , Glicoproteínas/genética , Pescoço/anormalidades , Doenças Faríngeas/genética , Região Branquial/cirurgia , Anormalidades Craniofaciais/cirurgia , Feminino , Mutação da Fase de Leitura , Avós , Humanos , Mutação INDEL , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Oxigenases de Função Mista/genética , Pescoço/cirurgia , Doenças Faríngeas/cirurgia , Polimorfismo de Nucleotídeo Único , Sequenciamento Completo do Exoma
4.
Nat Commun ; 11(1): 1092, 2020 02 27.
Artigo em Inglês | MEDLINE | ID: mdl-32107390

RESUMO

Micro(mi)RNA-based post-transcriptional regulatory mechanisms have been broadly implicated in the assembly and modulation of synaptic connections required to shape neural circuits, however, relatively few specific miRNAs have been identified that control synapse formation. Using a conditional transgenic toolkit for competitive inhibition of miRNA function in Drosophila, we performed an unbiased screen for novel regulators of synapse morphogenesis at the larval neuromuscular junction (NMJ). From a set of ten new validated regulators of NMJ growth, we discovered that miR-34 mutants display synaptic phenotypes and cell type-specific functions suggesting distinct downstream mechanisms in the presynaptic and postsynaptic compartments. A search for conserved downstream targets for miR-34 identified the junctional receptor CNTNAP4/Neurexin-IV (Nrx-IV) and the membrane cytoskeletal effector Adducin/Hu-li tai shao (Hts) as proteins whose synaptic expression is restricted by miR-34. Manipulation of miR-34, Nrx-IV or Hts-M function in motor neurons or muscle supports a model where presynaptic miR-34 inhibits Nrx-IV to influence active zone formation, whereas, postsynaptic miR-34 inhibits Hts to regulate the initiation of bouton formation from presynaptic terminals.


Assuntos
Proteínas de Ligação a Calmodulina/genética , Moléculas de Adesão Celular Neuronais/genética , Proteínas de Drosophila/genética , Regulação da Expressão Gênica no Desenvolvimento , MicroRNAs/metabolismo , Terminações Pré-Sinápticas/fisiologia , Animais , Animais Geneticamente Modificados , Proteínas de Ligação a Calmodulina/metabolismo , Moléculas de Adesão Celular Neuronais/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/fisiologia , Larva/crescimento & desenvolvimento , Morfogênese/genética , Mutação , Junção Neuromuscular/citologia , Junção Neuromuscular/crescimento & desenvolvimento
5.
Proc Natl Acad Sci U S A ; 117(1): 656-667, 2020 01 07.
Artigo em Inglês | MEDLINE | ID: mdl-31754030

RESUMO

A major challenge facing the genetics of autism spectrum disorders (ASDs) is the large and growing number of candidate risk genes and gene variants of unknown functional significance. Here, we used Caenorhabditis elegans to systematically functionally characterize ASD-associated genes in vivo. Using our custom machine vision system, we quantified 26 phenotypes spanning morphology, locomotion, tactile sensitivity, and habituation learning in 135 strains each carrying a mutation in an ortholog of an ASD-associated gene. We identified hundreds of genotype-phenotype relationships ranging from severe developmental delays and uncoordinated movement to subtle deficits in sensory and learning behaviors. We clustered genes by similarity in phenomic profiles and used epistasis analysis to discover parallel networks centered on CHD8•chd-7 and NLGN3•nlg-1 that underlie mechanosensory hyperresponsivity and impaired habituation learning. We then leveraged our data for in vivo functional assays to gauge missense variant effect. Expression of wild-type NLG-1 in nlg-1 mutant C. elegans rescued their sensory and learning impairments. Testing the rescuing ability of conserved ASD-associated neuroligin variants revealed varied partial loss of function despite proper subcellular localization. Finally, we used CRISPR-Cas9 auxin-inducible degradation to determine that phenotypic abnormalities caused by developmental loss of NLG-1 can be reversed by adult expression. This work charts the phenotypic landscape of ASD-associated genes, offers in vivo variant functional assays, and potential therapeutic targets for ASD.


Assuntos
Transtorno do Espectro Autista/genética , Moléculas de Adesão Celular Neuronais/genética , Habituação Psicofisiológica/genética , Fenômica/métodos , Animais , Animais Geneticamente Modificados , Transtorno do Espectro Autista/fisiopatologia , Técnicas de Observação do Comportamento/métodos , Comportamento Animal/fisiologia , Caenorhabditis elegans , Proteínas de Ligação a DNA/genética , Modelos Animais de Doenças , Epistasia Genética , Humanos , Imunoglobulinas/genética , Locomoção/genética , Proteínas de Membrana/genética , Mutação de Sentido Incorreto , Fenótipo , Fatores de Transcrição/genética
6.
Chemistry ; 26(8): 1834-1845, 2020 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-31773792

RESUMO

Chemical chaperones prevent protein aggregation. However, the use of chemical chaperones as drugs against diseases due to protein aggregation is limited by the very high active concentrations (mm range) required to mediate their effect. One of the most common chemical chaperones is 4-phenylbutyric acid (4-PBA). Despite its unfavorable pharmacokinetic properties, 4-PBA was approved as a drug to treat ornithine cycle diseases. Here, we report that 2-isopropyl-4-phenylbutanoic acid (5) has been found to be 2-10-fold more effective than 4-PBA in several in vitro models of protein aggregation. Importantly, compound 5 reduced the secretion rate of autism-linked Arg451Cys Neuroligin3 (R451C NLGN3).


Assuntos
Fenilbutiratos/química , Proteínas/química , Animais , Moléculas de Adesão Celular Neuronais/química , Moléculas de Adesão Celular Neuronais/genética , Moléculas de Adesão Celular Neuronais/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Células HEK293 , Humanos , Proteínas de Membrana/química , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Mutagênese Sítio-Dirigida , Proteínas do Tecido Nervoso/química , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/metabolismo , Células PC12 , Fenilbutiratos/farmacologia , Agregados Proteicos/efeitos dos fármacos , Dobramento de Proteína , Proteínas/metabolismo , Ratos
7.
Am J Pathol ; 190(2): 358-371, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31783007

RESUMO

Liver sinusoidal endothelial cells (LSECs) control organ functions, metabolism, and development through the secretion of angiokines. LSECs express hepatocyte growth factor (Hgf), which is involved in prenatal development, metabolic homeostasis, and liver regeneration. This study aimed to elucidate the precise contribution of LSEC-derived Hgf in physiological homeostasis and liver regeneration. Stab2-iCretg/wt;Hgffl/fl (HgfΔLSEC) mice were generated to abrogate Hgf expression selectively in LSECs from early fetal development onwards, to study global development, metabolic and endothelial zonation, and organ functions as well as liver regeneration in response to 70% partial hepatectomy (PH). Although zonation and liver/body weight ratios were not altered, total body weight and total liver weight were reduced in HgfΔLSEC. Necrotic organ damage was more marked in HgfΔLSEC mice, and regeneration was delayed 72 hours after PH. This was associated with decreased hepatocyte proliferation at 48 hours after PH. Molecularly, HgfΔLSEC mice showed down-regulation of Hgf/c-Met signaling and decreased expression of Deptor in hepatocytes. In vitro knockdown of Deptor was associated with decreased proliferation. Therefore, angiocrine Hgf controls hepatocyte proliferation and susceptibility to necrosis after partial hepatectomy via the Hgf/c-Met axis involving Deptor to prevent excessive organ damage.


Assuntos
Tamanho Corporal , Proliferação de Células , Fator de Crescimento de Hepatócito/fisiologia , Hepatócitos/citologia , Hepatopatias/prevenção & controle , Regeneração Hepática , Organogênese/fisiologia , Animais , Moléculas de Adesão Celular Neuronais/fisiologia , Endotélio/citologia , Endotélio/metabolismo , Feminino , Hepatectomia , Hepatócitos/fisiologia , Homeostase , Hepatopatias/metabolismo , Hepatopatias/patologia , Masculino , Camundongos , Camundongos Knockout , Comunicação Parácrina , Transdução de Sinais
8.
Development ; 147(2)2020 01 22.
Artigo em Inglês | MEDLINE | ID: mdl-31862845

RESUMO

The development of tissues and organs requires close interaction of cells. To achieve this, cells express adhesion proteins such as the neural cell adhesion molecule (NCAM) or its Drosophila ortholog Fasciclin 2 (Fas2). Both are members of the Ig-domain superfamily of proteins that mediate homophilic adhesion. These proteins are expressed as isoforms differing in their membrane anchorage and their cytoplasmic domains. To study the function of single isoforms, we have conducted a comprehensive genetic analysis of F as2 We reveal the expression pattern of all major Fas2 isoforms, two of which are GPI anchored. The remaining five isoforms carry transmembrane domains with variable cytoplasmic tails. We generated F as2 mutants expressing only single isoforms. In contrast to the null mutation, which causes embryonic lethality, these mutants are viable, indicating redundancy among the different isoforms. Cell type-specific rescue experiments showed that glial-secreted Fas2 can rescue the F as2 mutant phenotype to viability. This demonstrates that cytoplasmic Fas2 domains have no apparent essential functions and indicate that Fas2 has function(s) other than homophilic adhesion. In conclusion, our data suggest novel mechanistic aspects of a long-studied adhesion protein.


Assuntos
Moléculas de Adesão Celular Neuronais/metabolismo , Drosophila melanogaster/citologia , Drosophila melanogaster/metabolismo , Transdução de Sinais , Animais , Adesão Celular , Moléculas de Adesão Celular Neuronais/química , Moléculas de Adesão Celular Neuronais/genética , Movimento Celular , Drosophila melanogaster/embriologia , Drosophila melanogaster/genética , Embrião não Mamífero/citologia , Embrião não Mamífero/metabolismo , Edição de Genes , Regulação da Expressão Gênica no Desenvolvimento , Glicosilfosfatidilinositóis/metabolismo , Mutação/genética , Neuroglia/metabolismo , Domínios Proteicos , Isoformas de Proteínas/metabolismo , Traqueia/embriologia , Traqueia/metabolismo
9.
Methods Mol Biol ; 2043: 93-104, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31463905

RESUMO

Reelin is a large secreted protein that is essential for the brain development and function. Reelin is negatively regulated by the specific cleavage by a disintegrin and metalloproteinase with thrombospondin type 1 motifs 3 (ADAMTS-3) which is also secreted from neurons. It is likely that there are other proteases that can cleave Reelin. This chapter describes the protocol for expression and handling of recombinant Reelin and ADAMTS-3 proteins to facilitate investigation of these proteins.


Assuntos
Proteínas ADAMTS/genética , Moléculas de Adesão Celular Neuronais/genética , Proteínas da Matriz Extracelular/genética , Expressão Gênica , Proteínas do Tecido Nervoso/genética , Pró-Colágeno N-Endopeptidase/genética , Serina Endopeptidases/genética , Proteínas ADAMTS/metabolismo , Moléculas de Adesão Celular Neuronais/metabolismo , Proteínas da Matriz Extracelular/metabolismo , Células HEK293 , Humanos , Proteínas do Tecido Nervoso/metabolismo , Pró-Colágeno N-Endopeptidase/metabolismo , Engenharia de Proteínas , Proteínas Recombinantes/metabolismo , Serina Endopeptidases/metabolismo
10.
Methods Mol Biol ; 2043: 105-111, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31463906

RESUMO

Proteolytic cleavage of the secreted signaling protein Reelin has been suggested to play causative roles in many neuropsychiatric and neurodegenerative disorders. Therefore, characterization of the proteolytic activity against Reelin is important not only for understanding how the brain works but also for the development of novel therapy for these disorders. Notably, ADAMTS family proteases are the primary suspects of Reelin-cleaving proteases under many, though not all, circumstances. Here we describe how to measure the Reelin-cleaving activity of ADAMTS (or of any other protease that may cleave Reelin), how to purify the Reelin-cleaving protease ADAMTS-3 from the culture supernatant of cortical neurons, and how to detect endogenous Reelin protein and its fragments in the brain.


Assuntos
Proteínas ADAMTS/metabolismo , Encéfalo/metabolismo , Moléculas de Adesão Celular Neuronais/química , Córtex Cerebral/citologia , Proteínas da Matriz Extracelular/química , Proteínas do Tecido Nervoso/química , Pró-Colágeno N-Endopeptidase/metabolismo , Serina Endopeptidases/química , Animais , Células Cultivadas , Córtex Cerebral/metabolismo , Células HEK293 , Humanos , Camundongos , Neurônios/citologia , Neurônios/metabolismo , Proteólise
11.
Sci Total Environ ; 700: 134492, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31627046

RESUMO

Graphene oxide (GO) is a carbon-based engineered nanomaterial (ENM). Using Caenorhabditis elegans as an animal model, we investigated the effect of GO exposure on protein-protein interactions. In nematodes, NLG-1/Neuroligin, a postsynaptic protein, acted only in the neurons to regulate the GO toxicity. In the neurons, DLG-1, a PSD-95 protein, and MAGI-1, a S-SCAM protein, were identified as the downstream targets of NLG-1 in the regulation of GO toxicity. PKC-1, a serine/threonine protein kinase C, further acted downstream of neuronal DLG-1 and MAGI-1 to regulate the GO toxicity. Co-immunoprecipitation analysis demonstrated the protein-protein interaction between NLG-1 and DLG-1 or MAGI-1. After GO expression, this protein-protein interaction between NLG-1 and DLG-1 or MAGI-1 was significantly inhibited. Therefore, our data raised the evidence to suggest the potential of GO exposure in disrupting protein-protein interactions in organisms.


Assuntos
Proteínas de Caenorhabditis elegans/metabolismo , Caenorhabditis elegans/fisiologia , Grafite/toxicidade , Substâncias Perigosas/toxicidade , Animais , Moléculas de Adesão Celular Neuronais/metabolismo , Guanilato Quinases
12.
Biol Sex Differ ; 10(1): 62, 2019 12 18.
Artigo em Inglês | MEDLINE | ID: mdl-31852540

RESUMO

BACKGROUND: Since the early days of PCR techniques, sex identification, "sex-typing," of genomic DNA samples has been a fundamental part of human forensic analysis but also in animal genetics aiming at strategic livestock breeding. Most analyses are employing the AMELX/AMELY gene loci on the X and Y chromosomes present in most mammals. We hypothesize that sex-typing in humans is also possible based on the genes NLGN4X and NLGN4Y, which represent X and Y chromosome-specific copies of a common ancestral neuroligin-4 orthologue. METHODS: Genomic DNA was isolated from human blood and buccal cell samples (total n = 111) and submitted to two different strategies: (a) a traditional two-primer PCR approach detecting an insertion/deletion (indel) polymorphism immediately upstream of the translational start on exon 1 and (b) detection of a single nucleotide polymorphism, SNP, on the translational stop carrying exon 7. The SNP detection was based on a quantitative PCR approach (rhAMP genotyping) employing DNA/RNA hybrid oligonucleotides that were blocked and which could only be activated upon perfect annealing to the target DNA sequence. RESULTS: All indel PCR-tested human DNA samples showed two bands for males representing X- and Y-specific copies of NLGN4 and a single band for female samples, i.e., homozygosity of NLGN4X and absence of NLGN4Y, in accordance with the self-reported sex of the donors. These results were in perfect agreement with the results of the rhAMP-based SNP-detection method: all males were consequently positive for both alleles, representing either SNP variant, and females were interpreted as homozygous regarding the SNP variant found in NLGN4X. Both methods have shown reliable and consistent results that enabled us to infer the sex of donor DNA samples across different ethnicities. CONCLUSIONS: These results indicate that the detection of human NLGN4X/Y is a suitable alternative to previously reported methods employing gene loci such as AMELX/Y. Furthermore, this is the first report applying successfully the rhAMP-genotyping strategy as a means for SNP-based sex-typing, which consequently will be applicable to other gene loci or different species as well.


Assuntos
Moléculas de Adesão Celular Neuronais/genética , Análise para Determinação do Sexo , Transtorno Autístico/genética , Feminino , Humanos , Masculino , Reação em Cadeia da Polimerase/métodos , Polimorfismo de Nucleotídeo Único
13.
Neurol India ; 67(6): 1469-1471, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31857537

RESUMO

Objective: To study the DNA methylation profiles in brain tissue of patients with refractory epilepsy due to malformations of cortical development (MCDs). Materials and Methods: Clinical, neuroimaging, and pathology characteristics were defined for 13 patients who underwent resective surgery for epilepsy. Methylation analysis was performed using Illumina® 450k Methylation Microarray. Data analysis was completed, and pathway identification was done using the R/Bioconductor package. Results: Genes associated with Ephrin-Reelin pathway, potassium channels, and glutathione metabolism were differentially methylated in the MCD group when compared with patients who had no evidence of MCD. Conclusions: Our preliminary data reveal that epigenetic pathways may have a role in the pathobiogenesis of MCDs.


Assuntos
Encéfalo/metabolismo , Metilação de DNA , Epilepsia/genética , Malformações do Desenvolvimento Cortical/genética , Encéfalo/diagnóstico por imagem , Encéfalo/cirurgia , Moléculas de Adesão Celular Neuronais/genética , Moléculas de Adesão Celular Neuronais/metabolismo , Criança , Pré-Escolar , Epilepsia/etiologia , Epilepsia/cirurgia , Proteínas da Matriz Extracelular/genética , Proteínas da Matriz Extracelular/metabolismo , Feminino , Humanos , Lactente , Masculino , Malformações do Desenvolvimento Cortical/complicações , Malformações do Desenvolvimento Cortical/cirurgia , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/metabolismo , Projetos Piloto , Canais de Potássio/genética , Canais de Potássio/metabolismo , Serina Endopeptidases/genética , Serina Endopeptidases/metabolismo , Transdução de Sinais/genética
14.
EBioMedicine ; 50: 178-190, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31727599

RESUMO

BACKGROUND: Prostate cancer (PCa) is a leading cause of cancer-related death in males. Aberrant expression of long noncoding RNAs (lncRNAs) is frequently reported in human malignancies. This study was performed to explore the role of LSAMP-AS1 in epithelial-mesenchymal transition (EMT), proliferation, migration and invasion of PCa cells. METHODS: Initially, the differentially expressed lncRNAs in PCa were screened out by microarray analysis. The clinicopathological and prognostic significance of LSAMP-AS1 was evaluated. LSAMP-AS1 was over-expressed or silenced to investigate the roles in EMT, proliferation, migration and invasion of PCa cells. Moreover, the relationships between LSAMP-AS1 and miR-183-5p, as well as miR-183-5p and decorin (DCN) were characterized. The tumorigenicity of PCa cells was verified in nude mice. RESULTS: LSAMP-AS1 was poorly expressed in PCa tissues and cells. Low expression of LSAMP-AS1 was indicative of poor overall survival and disease-free survival, and related to Gleason score, TNM stage, and risk stratification. Over-expressed LSAMP-AS1 inhibited EMT, proliferation, migration and invasion of PCa cells, as well as tumor growth in nude mice. Meanwhile, over-expression of LSAMP-AS1 resulted in up-regulation of E-cadherin and down-regulation of Vimentin, N-cadherin, Ki67, PCNA, MMP-2, MMP-9, Ezrin and Fascin. Notably, LSAMP-AS1 competitively bound to miR-183-5p which directly targets DCN. It was confirmed that the inhibitory effect of LSAMP-AS1 on PCa cells was achieved by binding to miR-183-5p, thus promoting the expression of DCN. CONCLUSION: LSAMP-AS1 up-regulates the DCN gene by competitively binding to miR-183-5p, thus inhibiting EMT, proliferation, migration and invasion of PCa cells.


Assuntos
Moléculas de Adesão Celular Neuronais/genética , Decorina/genética , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , Neoplasias da Próstata/genética , Neoplasias da Próstata/patologia , Interferência de RNA , RNA Antissenso/genética , Idoso , Idoso de 80 Anos ou mais , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Progressão da Doença , Transição Epitelial-Mesenquimal/genética , Proteínas Ligadas por GPI/genética , Genes Reporter , Humanos , Masculino , Pessoa de Meia-Idade , Modelos Biológicos , Gradação de Tumores , Estadiamento de Neoplasias , Neoplasias da Próstata/mortalidade
15.
Yakugaku Zasshi ; 139(11): 1397-1402, 2019.
Artigo em Japonês | MEDLINE | ID: mdl-31685736

RESUMO

Patients with epilepsy are often affected by not only seizures but also a variety of cognitive and psychiatric comorbidities that further impair their quality of life. However, it is unclear whether epilepsy is associated with psychic function. The aim of the present study was to clarify the effects of kindling-induced epileptic seizures on psychic functioning, using behavioral pharmacological tests. Pentylenetetrazol (PTZ)-kindled mice displayed impaired motor coordination (in the rotarod test), and social approach impairment (in the three-chamber social test) compared with vehicle mice. Intraperitoneal ABT-418 treatment (0.05 mg/kg) alleviated these behavioral abnormalities in PTZ-kindled mice. Immunolabeling of tissue sections demonstrated that expression of the α4 subunit of the α4ß2 nicotinic acetylcholine receptor in the piriform cortex was significantly decreased in PTZ-kindled mice. In contrast, expression of the synaptic adhesion molecule neuroligin 3 (NLG3) was significantly higher in the piriform cortex of PTZ-kindled mice compared with vehicle mice. Collectively, our findings suggest that attention deficit/hyperactivity disorder (ADHD)-like or autistic-like behavioral abnormalities associated with epilepsy are closely linked to downregulation of the α4 subunit of the α4ß2 receptor and upregulation of NLG3 in the mouse piriform cortex. In summary, this study indicates that ABT-418 is a good candidate for the treatment of patients with epilepsy complicated by psychiatric symptoms such as autism and ADHD.


Assuntos
Epilepsia/psicologia , Excitação Neurológica , Transtornos Mentais/tratamento farmacológico , Transtornos Mentais/etiologia , Pentilenotetrazol , Receptores Nicotínicos/metabolismo , Convulsões/psicologia , Animais , Moléculas de Adesão Celular Neuronais/metabolismo , Modelos Animais de Doenças , Regulação para Baixo , Epilepsia/induzido quimicamente , Epilepsia/tratamento farmacológico , Epilepsia/etiologia , Isoxazóis/farmacologia , Isoxazóis/uso terapêutico , Proteínas de Membrana/metabolismo , Camundongos , Proteínas do Tecido Nervoso/metabolismo , Córtex Piriforme/metabolismo , Desempenho Psicomotor/efeitos dos fármacos , Pirrolidinas/farmacologia , Pirrolidinas/uso terapêutico , Qualidade de Vida , Convulsões/induzido quimicamente , Convulsões/tratamento farmacológico , Convulsões/etiologia , Comportamento Social
16.
Nat Commun ; 10(1): 4794, 2019 10 22.
Artigo em Inglês | MEDLINE | ID: mdl-31641127

RESUMO

Central nervous system myelin is a multilayered membrane produced by oligodendrocytes to increase neural processing speed and efficiency, but the molecular mechanisms underlying axonal selection and myelin wrapping are unknown. Here, using combined morphological and molecular analyses in mice and zebrafish, we show that adhesion molecules of the paranodal and the internodal segment work synergistically using overlapping functions to regulate axonal interaction and myelin wrapping. In the absence of these adhesive systems, axonal recognition by myelin is impaired with myelin growing on top of previously myelinated fibers, around neuronal cell bodies and above nodes of Ranvier. In addition, myelin wrapping is disturbed with the leading edge moving away from the axon and in between previously formed layers. These data show how two adhesive systems function together to guide axonal ensheathment and myelin wrapping, and provide a mechanistic understanding of how the spatial organization of myelin is achieved.


Assuntos
Axônios/fisiologia , Sistema Nervoso Central/fisiologia , Bainha de Mielina/fisiologia , Moléculas de Adesão de Célula Nervosa/metabolismo , Animais , Animais Geneticamente Modificados , Adesão Celular/fisiologia , Moléculas de Adesão Celular Neuronais/genética , Moléculas de Adesão Celular Neuronais/metabolismo , Contactina 1/genética , Contactina 1/metabolismo , Feminino , Larva , Masculino , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Bainha de Mielina/patologia , Glicoproteína Associada a Mielina/genética , Glicoproteína Associada a Mielina/metabolismo , Moléculas de Adesão de Célula Nervosa/genética , Nervo Óptico/metabolismo , Nervo Óptico/patologia , Peixe-Zebra/genética , Proteínas de Peixe-Zebra/genética , Proteínas de Peixe-Zebra/metabolismo
17.
Elife ; 82019 10 02.
Artigo em Inglês | MEDLINE | ID: mdl-31577229

RESUMO

The functions of FGF receptors (FGFRs) in early development of the cerebral cortex are well established. Their functions in the migration of neocortical projection neurons, however, are unclear. We have found that FGFRs regulate multipolar neuron orientation and the morphological change into bipolar cells necessary to enter the cortical plate. Mechanistically, our results suggest that FGFRs are activated by N-Cadherin. N-Cadherin cell-autonomously binds FGFRs and inhibits FGFR K27- and K29-linked polyubiquitination and lysosomal degradation. Accordingly, FGFRs accumulate and stimulate prolonged Erk1/2 phosphorylation. Neurons inhibited for Erk1/2 are stalled in the multipolar zone. Moreover, Reelin, a secreted protein regulating neuronal positioning, prevents FGFR degradation through N-Cadherin, causing Erk1/2 phosphorylation. These findings reveal novel functions for FGFRs in cortical projection neuron migration, suggest a physiological role for FGFR and N-Cadherin interaction in vivo and identify Reelin as an extracellular upstream regulator and Erk1/2 as downstream effectors of FGFRs during neuron migration.


Assuntos
Caderinas/metabolismo , Neocórtex/embriologia , Neurogênese , Neurônios/metabolismo , Receptores de Fatores de Crescimento de Fibroblastos/metabolismo , Ubiquitinação , Animais , Moléculas de Adesão Celular Neuronais/metabolismo , Proteínas da Matriz Extracelular/metabolismo , Sistema de Sinalização das MAP Quinases , Camundongos , Proteínas do Tecido Nervoso/metabolismo , Fosforilação , Serina Endopeptidases/metabolismo
18.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 36(9): 935-937, 2019 Sep 10.
Artigo em Chinês | MEDLINE | ID: mdl-31515794

RESUMO

OBJECTIVE: To explore the genetic basis for a patient with autism. METHODS: High-throughput sequencing was carried out to detect copy number variations in the patient. RESULTS: DNA sequencing found that the patient has carried a 0.11 Mb deletion in distal 2p16.3 spanning from genomic position 50 820 001 to 50 922 000, which resulted removal of exon 6 and part of intron 7 of the NRXN1 gene. The same deletion was not found his parents and brother. CONCLUSION: Partial deletion of the NRXN1 gene may underlie the disease in this patient.


Assuntos
Transtorno Autístico/genética , Moléculas de Adesão Celular Neuronais/genética , Deleção de Genes , Proteínas do Tecido Nervoso/genética , Variações do Número de Cópias de DNA , Humanos , Masculino
19.
Gene Expr Patterns ; 34: 119070, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31521773

RESUMO

Engulfment and cell motility (ELMO) proteins bind to Dock180, a guanine nucleotide exchange factor (GEF) of the Rac family, and regulate GEF activity. The resultant ELMO/Dock180/Rac module regulates cytoskeletal reorganization responsible for the engulfment of apoptotic cells, cell migration, and neurite extension. The expression and function of Elmo family proteins in the nervous system, however, are not yet fully understood. Here, we characterize the comparative gene expression profiles of three Elmo family members (Elmo1, Elmo2, and Elmo3) in the brain of C57BL/6J mice, a widely used inbred strain, together with reeler mutant mice to understand gene expression in normal laminated brain areas compared with abnormal areas. Although all three Elmo genes showed widespread mRNA expression over various mouse tissues tested, Elmo1 and Elmo2 were the major types expressed in the brain, and three Elmo genes were up-regulated between the first postnatal week (infant stage) and the third postnatal week (juvenile, weaning stage). In addition, the mRNAs of Elmo genes showed distinct distribution patterns in various brain areas and cell-types; such as neurons including inhibitory interneurons as well as some non-neuronal cells. In the cerebral cortex, the three Elmo genes were widely expressed over many cortical regions, but the predominant areas of Elmo1 and Elmo2 expression tended to be distributed unevenly in the deep (a lower part of the VI) and superficial (II/III) layers, respectively, which also changed depending on the cortical areas and postnatal stages. In the dentate gyrus of the hippocampus, Elmo2 was expressed in dentate granule cells more in the mature stage rather than the immature-differentiating stage. In the thalamus, Elmo1 but not the other members was highly expressed in many nuclei. In the medial habenula, Elmo2 and Elmo3 were expressed at intermediate levels. In the cerebellar cortex, Elmo1 and Elmo2 were expressed in differentiating-mature granule cells and mature granule cells, respectively. In the Purkinje cell layer, Elmo1 and Elmo2 were expressed in Purkinje cells and Bergmann glia, respectively. Disturbed cellular distributions and laminar structures caused by the reeler mutation did not severely change expression in these cell types despite the disturbed cellular distributions and laminar structures, including those of the cerebrum, hippocampus, and cerebellum. Taken together, these results suggested that these three Elmo family members share their functional roles in various brain regions during prenatal-postnatal development.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/genética , Proteínas do Citoesqueleto/genética , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Animais , Encéfalo/metabolismo , Moléculas de Adesão Celular Neuronais/metabolismo , Proliferação de Células , Proteínas do Citoesqueleto/metabolismo , Proteínas da Matriz Extracelular/metabolismo , Perfilação da Expressão Gênica/métodos , Fatores de Troca do Nucleotídeo Guanina/genética , Camundongos , Camundongos Endogâmicos C57BL , Proteínas do Tecido Nervoso/metabolismo , Serina Endopeptidases/metabolismo , Transdução de Sinais , Fatores de Transcrição/metabolismo , Transcriptoma/genética
20.
eNeuro ; 6(5)2019.
Artigo em Inglês | MEDLINE | ID: mdl-31540999

RESUMO

Bipolar disorder (BP) and schizophrenia (SCZ) are major psychiatric disorders, but the molecular mechanisms underlying the complicated pathologies of these disorders remain unclear. It is difficult to establish adequate in vitro models for pathological analysis because of the heterogeneity of these disorders. In the present study, to recapitulate the pathologies of these disorders in vitro, we established in vitro models by differentiating mature neurons from human induced pluripotent stem cells (hiPSCs) derived from BP and SCZ patient with contributive copy number variations, as follows: two BP patients with PCDH15 deletion and one SCZ patient with RELN deletion. Glutamatergic neurons and GABAergic neurons were induced from hiPSCs under optimized conditions. Both types of induced neurons from both hiPSCs exhibited similar phenotypes of MAP2 (microtubule-associated protein 2)-positive dendrite shortening and decreasing synapse numbers. Additionally, we analyzed isogenic PCDH15- or RELN-deleted cells. The dendrite and synapse phenotypes of isogenic neurons were partially similar to those of patient-derived neurons. These results suggest that the observed phenotypes are general phenotypes of psychiatric disorders, and our in vitro models using hiPSC-based technology may be suitable for analysis of the pathologies of psychiatric disorders.


Assuntos
Transtorno Bipolar/patologia , Técnicas de Cultura de Células/métodos , Neurônios/patologia , Células-Tronco Pluripotentes , Esquizofrenia/patologia , Adulto , Transtorno Bipolar/genética , Caderinas/genética , Moléculas de Adesão Celular Neuronais/genética , Células Cultivadas , Variações do Número de Cópias de DNA , Proteínas da Matriz Extracelular/genética , Feminino , Humanos , Técnicas In Vitro , Pessoa de Meia-Idade , Proteínas do Tecido Nervoso/genética , Esquizofrenia/genética , Serina Endopeptidases/genética
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