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Pesquisa : A11.872.700.250.750 [Categoria DeCS]
Referências encontradas : 605 [refinar]
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[PMID]:29220646
[Au] Autor:Hu P; Fabyanic E; Kwon DY; Tang S; Zhou Z; Wu H
[Ad] Endereço:Department of Genetics, University of Pennsylvania, Philadelphia PA 19104, USA; Epigenetics Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia PA 19104, USA.
[Ti] Título:Dissecting Cell-Type Composition and Activity-Dependent Transcriptional State in Mammalian Brains by Massively Parallel Single-Nucleus RNA-Seq.
[So] Source:Mol Cell;68(5):1006-1015.e7, 2017 Dec 07.
[Is] ISSN:1097-4164
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Massively parallel single-cell RNA sequencing can precisely resolve cellular diversity in a high-throughput manner at low cost, but unbiased isolation of intact single cells from complex tissues such as adult mammalian brains is challenging. Here, we integrate sucrose-gradient-assisted purification of nuclei with droplet microfluidics to develop a highly scalable single-nucleus RNA-seq approach (sNucDrop-seq), which is free of enzymatic dissociation and nucleus sorting. By profiling ∼18,000 nuclei isolated from cortical tissues of adult mice, we demonstrate that sNucDrop-seq not only accurately reveals neuronal and non-neuronal subtype composition with high sensitivity but also enables in-depth analysis of transient transcriptional states driven by neuronal activity, at single-cell resolution, in vivo.
[Mh] Termos MeSH primário: Núcleo Celular/metabolismo
Córtex Cerebral/metabolismo
Sequenciamento de Nucleotídeos em Larga Escala
Neurônios/metabolismo
RNA/genética
Convulsões/genética
Análise de Sequência de RNA/métodos
Análise de Célula Única/métodos
Transcrição Genética
[Mh] Termos MeSH secundário: Animais
Núcleo Celular/patologia
Centrifugação com Gradiente de Concentração
Córtex Cerebral/patologia
Córtex Cerebral/fisiopatologia
Modelos Animais de Doenças
Células-Tronco Embrionárias Humanas/metabolismo
Seres Humanos
Cinética
Masculino
Camundongos
Camundongos Endogâmicos C57BL
Técnicas Analíticas Microfluídicas
Células NIH 3T3
Inibição Neural
Neurônios/patologia
Pentilenotetrazol
RNA/metabolismo
Convulsões/metabolismo
Convulsões/patologia
Convulsões/fisiopatologia
Transmissão Sináptica
Transfecção
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
63231-63-0 (RNA); WM5Z385K7T (Pentylenetetrazole)
[Em] Mês de entrada:1712
[Cu] Atualização por classe:180228
[Lr] Data última revisão:
180228
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171209
[St] Status:MEDLINE


  2 / 605 MEDLINE  
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[PMID]:28469040
[Au] Autor:Smith BN; Topp SD; Fallini C; Shibata H; Chen HJ; Troakes C; King A; Ticozzi N; Kenna KP; Soragia-Gkazi A; Miller JW; Sato A; Dias DM; Jeon M; Vance C; Wong CH; de Majo M; Kattuah W; Mitchell JC; Scotter EL; Parkin NW; Sapp PC; Nolan M; Nestor PJ; Simpson M; Weale M; Lek M; Baas F; Vianney de Jong JM; Ten Asbroek ALMA; Redondo AG; Esteban-Pérez J; Tiloca C; Verde F; Duga S; Leigh N; Pall H; Morrison KE; Al-Chalabi A; Shaw PJ; Kirby J; Turner MR; Talbot K; Hardiman O; Glass JD; De Belleroche J; Maki M; Moss SE; Miller C; Gellera C
[Ad] Endereço:United Kingdom Dementia Research Institute Centre, Maurice Wohl Clinical Neuroscience Institute, Institute of Psychiatry, Psychology and Neuroscience, King's College London, 125 Coldharbour Lane, Camberwell, SE5 9NU London, UK.
[Ti] Título:Mutations in the vesicular trafficking protein annexin A11 are associated with amyotrophic lateral sclerosis.
[So] Source:Sci Transl Med;9(388), 2017 May 03.
[Is] ISSN:1946-6242
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder. We screened 751 familial ALS patient whole-exome sequences and identified six mutations including p.D40G in the gene in 13 individuals. The p.D40G mutation was absent from 70,000 control whole-exome sequences. This mutation segregated with disease in two kindreds and was present in another two unrelated cases ( = 0.0102), and all mutation carriers shared a common founder haplotype. Annexin A11-positive protein aggregates were abundant in spinal cord motor neurons and hippocampal neuronal axons in an ALS patient carrying the p.D40G mutation. Transfected human embryonic kidney cells expressing with the p.D40G mutation and other N-terminal mutations showed altered binding to calcyclin, and the p.R235Q mutant protein formed insoluble aggregates. We conclude that mutations in are associated with ALS and implicate defective intracellular protein trafficking in disease pathogenesis.
[Mh] Termos MeSH primário: Esclerose Amiotrófica Lateral/genética
Anexinas/genética
[Mh] Termos MeSH secundário: Anexinas/metabolismo
Células-Tronco Embrionárias Humanas/metabolismo
Seres Humanos
Mutação/genética
Ligação Proteica
Transporte Proteico
Proteína A6 Ligante de Cálcio S100/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Annexins); 0 (S100 Calcium Binding Protein A6)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180220
[Lr] Data última revisão:
180220
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170505
[St] Status:MEDLINE


  3 / 605 MEDLINE  
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Barraviera, Benedito
Texto completo
[PMID]:28450050
[Au] Autor:Araújo MR; Kyrylenko S; Spejo AB; Castro MV; Ferreira Junior RS; Barraviera B; Oliveira ALR
[Ad] Endereço:Department of Structural and Functional Biology, Institute of Biology, University of Campinas, Campinas, Sao Paulo, Brazil.
[Ti] Título:Transgenic human embryonic stem cells overexpressing FGF2 stimulate neuroprotection following spinal cord ventral root avulsion.
[So] Source:Exp Neurol;294:45-57, 2017 08.
[Is] ISSN:1090-2430
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Ventral root avulsion (VRA) triggers a strong glial reaction which contributes to neuronal loss, as well as to synaptic detachment. To overcome the degenerative effects of VRA, treatments with neurotrophic factors and stem cells have been proposed. Thus, we investigated neuroprotection elicited by human embryonic stem cells (hESC), modified to overexpress a human fibroblast growth factor 2 (FGF-2), on motoneurons subjected to VRA. Lewis rats were submitted to VRA (L4-L6) and hESC/FGF-2 were applied to the injury site using a fibrin scaffold. The spinal cords were processed to evaluate neuronal survival, synaptic stability, and glial reactivity two weeks post lesion. Then, qRT-PCR was used to assess gene expression of ß2-microglobulin (ß2m), TNFα, IL1ß, IL6 and IL10 in the spinal cord in vivo and FGF2 mRNA levels in hESC in vitro. The results indicate that hESC overexpressing FGF2 significantly rescued avulsed motoneurons, preserving synaptic covering and reducing astroglial reactivity. The cells were also shown to express BDNF and GDNF at the site of injury. Additionally, engraftment of hESC led to a significant reduction in mRNA levels of TNFα at the spinal cord ventral horn, indicating their immunomodulatory properties. Overall, the present data suggest that hESC overexpressing FGF2 are neuroprotective and can shift gene expression towards an anti-inflammatory environment.
[Mh] Termos MeSH primário: Células-Tronco Embrionárias Humanas/transplante
Radiculopatia/cirurgia
Raízes Nervosas Espinhais/patologia
[Mh] Termos MeSH secundário: Animais
Movimento Celular
Sobrevivência Celular/efeitos dos fármacos
Sobrevivência Celular/genética
Modelos Animais de Doenças
Doxiciclina/uso terapêutico
Feminino
Adesivo Tecidual de Fibrina/toxicidade
Fator 2 de Crescimento de Fibroblastos/genética
Fator 2 de Crescimento de Fibroblastos/metabolismo
Regulação da Expressão Gênica/efeitos dos fármacos
Regulação da Expressão Gênica/genética
Vetores Genéticos/fisiologia
Células-Tronco Embrionárias Humanas/metabolismo
Seres Humanos
Neurônios Motores/metabolismo
Neurônios Motores/patologia
Proteínas do Tecido Nervoso/metabolismo
Neuroglia/efeitos dos fármacos
Neuroglia/metabolismo
Radiculopatia/induzido quimicamente
Ratos
Ratos Endogâmicos Lew
Adesivos Teciduais/toxicidade
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Fibrin Tissue Adhesive); 0 (Nerve Tissue Proteins); 0 (Tissue Adhesives); 103107-01-3 (Fibroblast Growth Factor 2); N12000U13O (Doxycycline)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:180218
[Lr] Data última revisão:
180218
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170429
[St] Status:MEDLINE


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[PMID]:28464615
[Au] Autor:Nygren H; Bigdeli N; Ilver L; Malmberg P
[Ad] Endereço:Department of Medical Chemistry and Cell Biology, Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg, POB 440, 40530 Gothenburg, Sweden.
[Ti] Título:Mg-corrosion, hydroxyapatite, and bone healing.
[So] Source:Biointerphases;12(2):02C407, 2017 05 02.
[Is] ISSN:1559-4106
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The different capacities of magnesium in the metallic form (Mg-metal) and magnesium oxide (MgO) to stimulate bone healing are possible clues in the search for products that may promote bone healing. Since both Mg-metal and MgO can be assumed to release comparable amounts of Mg ions during their reactions in the tissue where they have been implanted, it is of some importance to follow this process and analyze the resulting mineral formation in the tissue at the implantation site. Implants of MgO were inserted into rat tibia, and the bone healing was compared with sham-operated controls. Samples were taken after 1 week of healing and analyzed by histology, environmental scanning electron microscopy equipped with an energy dispersive x-ray spectroscopy analyzer, and time-of-flight secondary ion mass spectrometry (ToF-SIMS). Callus bone was seen in sham-operated controls after 1 week of healing. Implantation of MgO impaired the callus bone formation by replacing bone with apparently mineralized areas, lacking osteocytes and were denoted, amorphous bodies. Elemental analysis showed increased levels of Ca (7.1%), P (3.7%), and Mg (0.2%) in the bone marrow of MgO-treated animals versus sham-operated controls Ca (2.4%), P (2.3%), and Mg (0.1%). The Ca content of the cortical bone was also significantly increased (Ca, 29% increase) in MgO-treated animals compared to sham-operated controls. The Ca content of the cortical bone of sham-operated animals was also significantly (p < 0.05) higher than the corresponding value of untreated animals, which means that the surgical trauma induces an altered composition of the bone mineral. The Ca/P ratio was 1.26-1.68, which is compatible with that of mineralized bone with different contents of organic materials. Analysis of bone sections using ToF-SIMS showed the presence of hydroxyapatite (HA) and MgCO in the bone marrow and in cortical bone. Analysis using x-ray photoelectron spectroscopy of Mg, MgO, and MgCO after incubation with cell culture medium (DMEM), in vitro, showed binding of CaPO at the Mg and MgO samples. The Ca/P ratio was 0.8, indicating a higher P content than that expected for HA. Exposure of human embryonic stem cells to Mg species preincubated in DMEM resulted in HA production by the cells. Thus, two sources of CaPO in the bone marrow of MgO-treated bone were defined, catalytic formation on Mg-species and synthesis from activated stem-cells. The presented data suggest that bone healing near Mg implants is congruent with the fracture healing of bone, boosted by high HA levels in the bone marrow. In this context, the different capacities of Mg-metal and MgO to catalyse the formation of HA can be important clues to their different bone promoting effects.
[Mh] Termos MeSH primário: Calo Ósseo/metabolismo
Osso Cortical/lesões
Osso Cortical/metabolismo
Durapatita/farmacologia
Consolidação da Fratura/efeitos dos fármacos
Óxido de Magnésio/farmacologia
[Mh] Termos MeSH secundário: Animais
Calo Ósseo/patologia
Linhagem Celular
Corrosão
Osso Cortical/patologia
Células-Tronco Embrionárias Humanas
Seres Humanos
Magnésio/farmacologia
Masculino
Teste de Materiais
Ratos
Ratos Sprague-Dawley
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
3A3U0GI71G (Magnesium Oxide); 91D9GV0Z28 (Durapatite); I38ZP9992A (Magnesium)
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180201
[Lr] Data última revisão:
180201
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170504
[St] Status:MEDLINE
[do] DOI:10.1116/1.4982601


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[PMID]:29186714
[Au] Autor:Zhao HX; Jiang F; Zhu YJ; Wang L; Li K; Li Y; Wang XH; Li LS; Yao YQ
[Ad] Endereço:Department of Obstetrics and Gynecology, Tangdu Hospital, The Fourth Military Medical University, Xi'an, China.
[Ti] Título:Enhanced Immunological Tolerance by HLA-G1 from Neural Progenitor Cells (NPCs) Derived from Human Embryonic Stem Cells (hESCs).
[So] Source:Cell Physiol Biochem;44(4):1435-1444, 2017.
[Is] ISSN:1421-9778
[Cp] País de publicação:Switzerland
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Despite the great potential of utilizing human embryonic stem cells (hESCs)-derived cells as cell source for transplantation, these cells were often rejected during engraftment by the immune system due to adaptive immune response. METHODS: We first evaluated HLA-G expression level in both hESCs and differentiated progenitor cells. After that, we generated modified hESC lines that over-express HLA-G1 using lentiviral infection with the construct contains both HLA-G1 and GFP tag. The lentivirus was first produced by co-transfecting HLA-G1 expressing lentiviral vector together with packaging vectors into packaging cell line 293T. Then the produced virus was used for the infection of selected hESC lines. We characterized the generated cell lines phenotype, including pluripotency and self-renewal abilities, as well as immune tolerance ability by mixed lymphocyte reaction (MLR) and cytotoxicity assays. RESULTS: Although the hESCs do not express high levels of HLA-G1, over-expression of HLA-G1 in hESCs still retains their stem cell characteristics as determined by retaining the expression levels of OCT4 and SOX2, two critical transcriptional factors for stem cell function. Furthermore, the HLA-G1 overexpressing hESCs retain the self-renewal and pluripotency characteristics of stem cells, which can differentiate into different types of cells, including pigment cells, smooth muscle cells, epithelia-like cells, and NPCs. After differentiation, the differentiated cells including NPCs retain the high levels of HLA-G1 protein. In comparison with conventional NPCs, these HLA-G1 positive NPCs have enhanced immune tolerance ability. CONCLUSIONS: Ectopic expression of HLA-G1, a non-classical major histocompatibility complex class I (MHC I) antigen that was originally discovered involving in engraftment tolerance during pregnancy, can enhance the immunological tolerance in differentiated neural progenitor cells (NPCs). Our study shows that stably overexpressing HLA-G1 in hESCs might be a feasible strategy for enhancing the engraftment of NPCs during transplantation.
[Mh] Termos MeSH primário: Antígenos HLA-G/metabolismo
Tolerância Imunológica/fisiologia
Células-Tronco Neurais/metabolismo
[Mh] Termos MeSH secundário: Diferenciação Celular
Vetores Genéticos/genética
Vetores Genéticos/metabolismo
Antígenos HLA-G/genética
Células-Tronco Embrionárias Humanas/citologia
Células-Tronco Embrionárias Humanas/metabolismo
Seres Humanos
Cariótipo
Lentivirus/genética
Células-Tronco Neurais/citologia
Fator 3 de Transcrição de Octâmero/metabolismo
Fatores de Transcrição SOXB1/metabolismo
Teratoma/patologia
Transfecção
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (HLA-G Antigens); 0 (Octamer Transcription Factor-3); 0 (POU5F1 protein, human); 0 (SOX2 protein, human); 0 (SOXB1 Transcription Factors)
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180118
[Lr] Data última revisão:
180118
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171130
[St] Status:MEDLINE
[do] DOI:10.1159/000485539


  6 / 605 MEDLINE  
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[PMID]:27775225
[Au] Autor:Kamei KI; Koyama Y; Tokunaga Y; Mashimo Y; Yoshioka M; Fockenberg C; Mosbergen R; Korn O; Wells C; Chen Y
[Ad] Endereço:Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Kyoto, 6068501, Japan.
[Ti] Título:Characterization of Phenotypic and Transcriptional Differences in Human Pluripotent Stem Cells under 2D and 3D Culture Conditions.
[So] Source:Adv Healthc Mater;5(22):2951-2958, 2016 Nov.
[Is] ISSN:2192-2659
[Cp] País de publicação:Germany
[La] Idioma:eng
[Ab] Resumo:Human pluripotent stem cells hold great promise for applications in drug discovery and regenerative medicine. Microfluidic technology is a promising approach for creating artificial microenvironments; however, although a proper 3D microenvironment is required to achieve robust control of cellular phenotypes, most current microfluidic devices provide only 2D cell culture and do not allow tuning of physical and chemical environmental cues simultaneously. Here, the authors report a 3D cellular microenvironment plate (3D-CEP), which consists of a microfluidic device filled with thermoresponsive poly(N-isopropylacrylamide)-ß-poly(ethylene glycol) hydrogel (HG), which enables systematic tuning of both chemical and physical environmental cues as well as in situ cell monitoring. The authors show that H9 human embryonic stem cells (hESCs) and 253G1 human induced pluripotent stem cells in the HG/3D-CEP system maintain their pluripotent marker expression under HG/3D-CEP self-renewing conditions. Additionally, global gene expression analyses are used to elucidate small variations among different test environments. Interestingly, the authors find that treatment of H9 hESCs under HG/3D-CEP self-renewing conditions results in initiation of entry into the neural differentiation process by induction of PAX3 and OTX1 expression. The authors believe that this HG/3D-CEP system will serve as a versatile platform for developing targeted functional cell lines and facilitate advances in drug screening and regenerative medicine.
[Mh] Termos MeSH primário: Células-Tronco Pluripotentes/citologia
[Mh] Termos MeSH secundário: Resinas Acrílicas/administração & dosagem
Resinas Acrílicas/química
Biomarcadores/metabolismo
Técnicas de Cultura de Células/métodos
Diferenciação Celular/fisiologia
Células Cultivadas
Microambiente Celular/fisiologia
Células-Tronco Embrionárias Humanas/citologia
Seres Humanos
Hidrogel de Polietilenoglicol-Dimetacrilato/administração & dosagem
Hidrogel de Polietilenoglicol-Dimetacrilato/química
Dispositivos Lab-On-A-Chip
Células-Tronco Pluripotentes/metabolismo
Polietilenoglicóis/administração & dosagem
Polietilenoglicóis/química
Medicina Regenerativa/métodos
Transcrição Genética/fisiologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Acrylic Resins); 0 (Biomarkers); 25189-55-3 (poly-N-isopropylacrylamide); 25852-47-5 (Hydrogel, Polyethylene Glycol Dimethacrylate); 30IQX730WE (Polyethylene Glycols)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171130
[Lr] Data última revisão:
171130
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161025
[St] Status:MEDLINE
[do] DOI:10.1002/adhm.201600893


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[PMID]:28989025
[Au] Autor:Chan PKW; Geng L; Gao Y; Keung W; Li RA
[Ad] Endereço:Ming Wai Lau Centre for Reparative Medicine, Karolinska Institutet, Stockholm 17177, Sweden.
[Ti] Título:AAV-mediated conversion of human pluripotent stem cell-derived pacemaker.
[So] Source:Biochem Biophys Res Commun;494(1-2):346-351, 2017 Dec 09.
[Is] ISSN:1090-2104
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Malfunction of nodal pacemaker (Pm) cardiomyocytes (CMs) due to diseases or aging leads to rhythm generation disorders, necessitating electronic Pm implantation. We functionally reprogrammed human pluripotent stem cell (hPSC) derived-ventricular (V) CMs into -PmCMs via recombinant adeno-associated virus serotype 9 (rAAV9)-mediated overexpression of engineered HCN1 channel (HCN1ΔΔΔ) whose S3-S4 linker has been strategically deleted by design to promote cardiac pacemaking. rAAV9-HCN1ΔΔΔ-reprogrammed hPSC-PmCMs converted from -VCMs showed automaticity and action potential parameters typical of native nodal PmCMs. Implantation of rAAV9-HCN1ΔΔΔ-based BPm in a preclinical porcine model of complete heart block significantly reduced the dependence on device-supported pacing and generated spontaneous heart rhythms from the BPm. Collectively, these results have further laid the groundwork on BPm for future translation.
[Mh] Termos MeSH primário: Dependovirus/metabolismo
Bloqueio Cardíaco/terapia
Ventrículos do Coração/metabolismo
Canais Disparados por Nucleotídeos Cíclicos Ativados por Hiperpolarização/metabolismo
Miócitos Cardíacos/metabolismo
Células-Tronco Pluripotentes/metabolismo
Canais de Potássio/metabolismo
[Mh] Termos MeSH secundário: Potenciais de Ação/fisiologia
Animais
Diferenciação Celular
Reprogramação Celular
Dependovirus/genética
Modelos Animais de Doenças
Expressão Gênica
Genes Reporter
Vetores Genéticos/química
Vetores Genéticos/metabolismo
Proteínas de Fluorescência Verde/genética
Proteínas de Fluorescência Verde/metabolismo
Bloqueio Cardíaco/genética
Bloqueio Cardíaco/metabolismo
Bloqueio Cardíaco/fisiopatologia
Frequência Cardíaca/fisiologia
Ventrículos do Coração/patologia
Ventrículos do Coração/fisiopatologia
Células-Tronco Embrionárias Humanas/citologia
Células-Tronco Embrionárias Humanas/metabolismo
Seres Humanos
Canais Disparados por Nucleotídeos Cíclicos Ativados por Hiperpolarização/genética
Miócitos Cardíacos/citologia
Marca-Passo Artificial
Células-Tronco Pluripotentes/citologia
Canais de Potássio/genética
Suínos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (HCN1 protein, human); 0 (Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels); 0 (Potassium Channels); 147336-22-9 (Green Fluorescent Proteins)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171108
[Lr] Data última revisão:
171108
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171010
[St] Status:MEDLINE


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[PMID]:28953884
[Au] Autor:Fogarty NME; McCarthy A; Snijders KE; Powell BE; Kubikova N; Blakeley P; Lea R; Elder K; Wamaitha SE; Kim D; Maciulyte V; Kleinjung J; Kim JS; Wells D; Vallier L; Bertero A; Turner JMA; Niakan KK
[Ad] Endereço:Human Embryo and Stem Cell Laboratory, The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK.
[Ti] Título:Genome editing reveals a role for OCT4 in human embryogenesis.
[So] Source:Nature;550(7674):67-73, 2017 10 05.
[Is] ISSN:1476-4687
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Despite their fundamental biological and clinical importance, the molecular mechanisms that regulate the first cell fate decisions in the human embryo are not well understood. Here we use CRISPR-Cas9-mediated genome editing to investigate the function of the pluripotency transcription factor OCT4 during human embryogenesis. We identified an efficient OCT4-targeting guide RNA using an inducible human embryonic stem cell-based system and microinjection of mouse zygotes. Using these refined methods, we efficiently and specifically targeted the gene encoding OCT4 (POU5F1) in diploid human zygotes and found that blastocyst development was compromised. Transcriptomics analysis revealed that, in POU5F1-null cells, gene expression was downregulated not only for extra-embryonic trophectoderm genes, such as CDX2, but also for regulators of the pluripotent epiblast, including NANOG. By contrast, Pou5f1-null mouse embryos maintained the expression of orthologous genes, and blastocyst development was established, but maintenance was compromised. We conclude that CRISPR-Cas9-mediated genome editing is a powerful method for investigating gene function in the context of human development.
[Mh] Termos MeSH primário: Desenvolvimento Embrionário/genética
Edição de Genes
Regulação da Expressão Gênica no Desenvolvimento
Fator 3 de Transcrição de Octâmero/genética
Fator 3 de Transcrição de Octâmero/metabolismo
[Mh] Termos MeSH secundário: Animais
Blastocisto/metabolismo
Sistemas CRISPR-Cas/genética
Linhagem da Célula
Ectoderma/metabolismo
Embrião de Mamíferos/citologia
Embrião de Mamíferos/embriologia
Embrião de Mamíferos/metabolismo
Feminino
Camadas Germinativas/metabolismo
Células-Tronco Embrionárias Humanas/citologia
Células-Tronco Embrionárias Humanas/metabolismo
Seres Humanos
Masculino
Camundongos
Proteína Homeobox Nanog/genética
Proteína Homeobox Nanog/metabolismo
Fator 3 de Transcrição de Octâmero/deficiência
Especificidade por Substrato
Zigoto/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Nanog Homeobox Protein); 0 (Octamer Transcription Factor-3); 0 (POU5F1 protein, human)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171024
[Lr] Data última revisão:
171024
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170928
[St] Status:MEDLINE
[do] DOI:10.1038/nature24033


  9 / 605 MEDLINE  
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[PMID]:28943339
[Au] Autor:Sun H; Wang X; Liu K; Guo M; Zhang Y; Ying QL; Ye S
[Ad] Endereço:Center for Stem Cell and Translational Medicine, School of Life Sciences, Anhui University, Hefei 230601, PR China.
[Ti] Título:ß-catenin coordinates with Jup and the TCF1/GATA6 axis to regulate human embryonic stem cell fate.
[So] Source:Dev Biol;431(2):272-281, 2017 11 15.
[Is] ISSN:1095-564X
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:ß-catenin-mediated signaling has been extensively studied in regard to its role in the regulation of human embryonic stem cells (hESCs). However, the results are controversial and the mechanism by which ß-catenin regulates the hESC fate remains unclear. Here, we report that ß-catenin and γ-catenin are functionally redundant in mediating hESC adhesion and are required for embryoid body formation, but both genes are dispensable for hESC maintenance, as the undifferentiated state of ß-catenin and γ-catenin double deficient hESCs can be maintained. Overexpression of ß-catenin induces rapid hESC differentiation. Functional assays revealed that TCF1 plays a crucial role in hESC differentiation mediated by ß-catenin. Forced expression of TCF1, but not other LEF1/TCF family members, resulted in hESC differentiation towards the definitive endoderm. Conversely, knockdown of TCF1 or inhibition of the interaction between TCF1 and ß-catenin delayed hESC exit from pluripotency. Furthermore, we demonstrated that GATA6 plays a predominant role in TCF1-mediated hESC differentiation. Knockdown of GATA6 completely eliminated the effect of TCF1, while forced expression of GATA6 induced hESC differentiation. Our data thus reveal more detailed mechanisms for ß-catenin in regulating hESC fate decisions and will expand our understanding of the self-renewal and differentiation circuitry in hESCs.
[Mh] Termos MeSH primário: Linhagem da Célula
Fator de Transcrição GATA6/metabolismo
Células-Tronco Embrionárias Humanas/citologia
Células-Tronco Embrionárias Humanas/metabolismo
Fator 1 de Ligação ao Facilitador Linfoide/metabolismo
Transdução de Sinais
beta Catenina/metabolismo
[Mh] Termos MeSH secundário: Adesão Celular
Diferenciação Celular
Autorrenovação Celular
Desmoplaquinas/metabolismo
Endoderma/citologia
Seres Humanos
Transcrição Genética
Regulação para Cima
gama Catenina
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Desmoplakins); 0 (GATA6 Transcription Factor); 0 (JUP protein, human); 0 (LEF1 protein, human); 0 (Lymphoid Enhancer-Binding Factor 1); 0 (beta Catenin); 0 (gamma Catenin)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171116
[Lr] Data última revisão:
171116
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170926
[St] Status:MEDLINE


  10 / 605 MEDLINE  
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[PMID]:28911101
[Au] Autor:Deng Y; Bao F; Yang Y; Ji X; Du M; Zhang Z; Wang M; Dai Q
[Ad] Endereço:Department of Automation, Tsinghua National Laboratory for Information Science and Technology, Tsinghua University, Beijing 100084, P. R. China.
[Ti] Título:Information transduction capacity reduces the uncertainties in annotation-free isoform discovery and quantification.
[So] Source:Nucleic Acids Res;45(15):e143, 2017 Sep 06.
[Is] ISSN:1362-4962
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:The automated transcript discovery and quantification of high-throughput RNA sequencing (RNA-seq) data are important tasks of next-generation sequencing (NGS) research. However, these tasks are challenging due to the uncertainties that arise in the inference of complete splicing isoform variants from partially observed short reads. Here, we address this problem by explicitly reducing the inherent uncertainties in a biological system caused by missing information. In our approach, the RNA-seq procedure for transforming transcripts into short reads is considered an information transmission process. Consequently, the data uncertainties are substantially reduced by exploiting the information transduction capacity of information theory. The experimental results obtained from the analyses of simulated datasets and RNA-seq datasets from cell lines and tissues demonstrate the advantages of our method over state-of-the-art competitors. Our algorithm is an open-source implementation of MaxInfo.
[Mh] Termos MeSH primário: Algoritmos
Sequenciamento de Nucleotídeos em Larga Escala/métodos
Isoformas de Proteínas/genética
Processamento de RNA/genética
Análise de Sequência de RNA/métodos
[Mh] Termos MeSH secundário: Animais
Células Cultivadas
Biologia Computacional/métodos
Conjuntos de Dados como Assunto
Drosophila melanogaster/genética
Perfilação da Expressão Gênica/métodos
Células-Tronco Embrionárias Humanas/metabolismo
Seres Humanos
Isoformas de Proteínas/análise
RNA Mensageiro/análise
Software
Transcriptoma
[Pt] Tipo de publicação:EVALUATION STUDIES; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Protein Isoforms); 0 (RNA, Messenger)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171107
[Lr] Data última revisão:
171107
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170916
[St] Status:MEDLINE
[do] DOI:10.1093/nar/gkx585



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