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  1 / 3299 MEDLINE  
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[PMID]:29447173
[Au] Autor:Diao YF; Lin T; Li X; Oqani RK; Lee JE; Kim SY; Jin DI
[Ad] Endereço:Institute of Special Animal & Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun, China.
[Ti] Título:Dynamic changes of SETD2, a histone H3K36 methyltransferase, in porcine oocytes, IVF and SCNT embryos.
[So] Source:PLoS One;13(2):e0191816, 2018.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:SETD2 (SET domain containing protein 2) acts as a histone H3 lysine 36 (H3K36)-specific methyltransferase and may play important roles in active gene transcription in human cells. However, its expression and role in porcine oocytes and preimplantation embryos are not well understood. Here, we used immunofluorescence and laser scanning confocal microscopy to examine SETD2 expression in porcine fetal fibroblasts, oocytes, and preimplantation embryos derived from in vitro fertilization (IVF), parthenogenetic activation (PA), and somatic cell nuclear transfer (SCNT). In porcine fetal fibroblasts, SETD2 expression was detected in interphase cells, but not in M (mitotic)-phase cells. The SETD2 signal was observed in non-surrounded nucleolus (NSN)-stage oocytes, but not in surrounded nucleolus (SN)-, metaphase I (MI)-, or metaphase II (MII)-stage oocytes. The SETD2 signal was detectable in sperm, and undetectable immediately after fertilization, detectable at the 2-cell stage, and peaked at the 4-cell stage of IVF embryos in which porcine embryonic genome is activated. Similar to the pattern found in IVF embryos, the SETD2 signal was absent from PA embryos at the 1-cell stage, but it was detected at the 2-cell stage and thereafter maintained to the blastocyst stage. Interestingly, unlike the IVF and PA embryos, the SETD2 signal was detected throughout the development of SCNT embryos, including at the 1-cell stage. These data suggest that SETD2 may be functional for embryonic gene transcription in porcine preimplantation embryos. It is further speculated that the aberrant expression of SETD2 at the 1-cell stage of porcine SCNT embryos may be a factor in the low efficiency of cloning in pig.
[Mh] Termos MeSH primário: Fertilização In Vitro
Histona-Lisina N-Metiltransferase/metabolismo
Técnicas de Transferência Nuclear
Oócitos/enzimologia
[Mh] Termos MeSH secundário: Animais
Blastocisto
Células Cultivadas
Oócitos/citologia
Partenogênese
Suínos
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
EC 2.1.1.43 (Histone-Lysine N-Methyltransferase)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180309
[Lr] Data última revisão:
180309
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180216
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0191816


  2 / 3299 MEDLINE  
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[PMID]:28468955
[Au] Autor:Kaminuma O; Katayama K; Inoue K; Saeki M; Nishimura T; Kitamura N; Shimo Y; Tofukuji S; Ishida S; Ogonuki N; Kamimura S; Oikawa M; Katoh S; Mori A; Shichijo M; Hiroi T; Ogura A
[Ad] Endereço:Allergy and Immunology Project, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan osamuk@yamanashi.ac.jp ogura@rtc.riken.go.jp.
[Ti] Título:Hyper-reactive cloned mice generated by direct nuclear transfer of antigen-specific CD4 T cells.
[So] Source:EMBO Rep;18(6):885-893, 2017 Jun.
[Is] ISSN:1469-3178
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:T-cell receptor (TCR)-transgenic mice have been employed for evaluating antigen-response mechanisms, but their non-endogenous TCR might induce immune response differently than the physiologically expressed TCR Nuclear transfer cloning produces animals that retain the donor genotype in all tissues including germline and immune systems. Taking advantage of this feature, we generated cloned mice that carry endogenously rearranged TCR genes from antigen-specific CD4 T cells. We show that T cells of the cloned mice display distinct developmental pattern and antigen reactivity because of their endogenously pre-rearranged TCRα (rTα) and TCRß (rTß) alleles. These alleles were transmitted to the offspring, allowing us to establish a set of mouse lines that show chronic-type allergic phenotypes, that is, bronchial and nasal inflammation, upon local administrations of the corresponding antigens. Intriguingly, the existence of either rTα or rTß is sufficient to induce hypersensitivity. These cloned mice expressing intrinsic promoter-regulated antigen-specific TCR are a unique animal model with allergic predisposition for investigating CD4 T-cell-mediated pathogenesis and cellular commitment in immune diseases.
[Mh] Termos MeSH primário: Linfócitos T CD4-Positivos/imunologia
Hipersensibilidade/imunologia
Técnicas de Transferência Nuclear
Receptores de Antígenos de Linfócitos T/genética
[Mh] Termos MeSH secundário: Alelos
Animais
Antígenos/administração & dosagem
Antígenos/imunologia
Clonagem de Organismos
Modelos Animais de Doenças
Camundongos
Camundongos Transgênicos
Receptores de Antígenos de Linfócitos T/imunologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antigens); 0 (Receptors, Antigen, T-Cell)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180223
[Lr] Data última revisão:
180223
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170505
[St] Status:MEDLINE
[do] DOI:10.15252/embr.201643321


  3 / 3299 MEDLINE  
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[PMID]:28980654
[Au] Autor:Cyranoski D
[Ti] Título:Chinese scientists fix genetic disorder in cloned human embryos.
[So] Source:Nature;550(7674):15-16, 2017 10 02.
[Is] ISSN:1476-4687
[Cp] País de publicação:England
[La] Idioma:eng
[Mh] Termos MeSH primário: Clonagem de Organismos
Técnicas de Transferência Nuclear
[Mh] Termos MeSH secundário: Seres Humanos
[Pt] Tipo de publicação:JOURNAL ARTICLE; COMMENT
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171122
[Lr] Data última revisão:
171122
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171006
[St] Status:MEDLINE
[do] DOI:10.1038/nature.2017.22694


  4 / 3299 MEDLINE  
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[PMID]:28794155
[Au] Autor:Zhang J; Qu P; Zhou C; Liu X; Ma X; Wang M; Wang Y; Su J; Liu J; Zhang Y
[Ad] Endereço:From the Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, Shaanxi, China.
[Ti] Título:MicroRNA-125b is a key epigenetic regulatory factor that promotes nuclear transfer reprogramming.
[So] Source:J Biol Chem;292(38):15916-15926, 2017 Sep 22.
[Is] ISSN:1083-351X
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Somatic cell nuclear transfer (SCNT)-mediated reprogramming is a rapid, efficient, and sophisticated process that reprograms differentiated somatic cells to a pluripotent state. However, many factors in this elaborate reprogramming process remain largely unknown. Here, we report that the microRNA (miR) miR-125b is an important component of SCNT-mediated reprogramming. Luciferase reporter assay, quantitative PCR, and Western blotting demonstrated that miR-125b directly binds the 3'-untranslated region of SUV39H1, encoding the histone-lysine -methyltransferase SUV39H1, to down-regulate histone H3 lysine-9 tri-methylation (H3K9me3) in SCNT embryos. Furthermore, the miR-125b/SUV39H1 interaction induced loss of SUV39H1-mediated H3K9me3, caused heterochromatin relaxation, and promoted the development of SCNT embryos. Transcriptome analyses of SCNT blastomeres indicated that HNF1 homeobox B (HNF1B), a gene encoding a transcription factor downstream of and controlled by the miR-125b/SUV39H1 axis, is important for conferring developmental competence on preimplantation embryos. We conclude that miR-125b promotes SCNT-mediated nuclear reprogramming by targeting SUV39H1 to decrease the deposition of repressive H3K9me3 modifications.
[Mh] Termos MeSH primário: Epigênese Genética
MicroRNAs/genética
Técnicas de Transferência Nuclear
[Mh] Termos MeSH secundário: Animais
Sequência de Bases
Regulação da Expressão Gênica/genética
Células HEK293
Fator 1-beta Nuclear de Hepatócito/genética
Heterocromatina/metabolismo
Histonas/química
Histonas/metabolismo
Seres Humanos
Lisina/metabolismo
Metilação
Metiltransferases/genética
Camundongos
Proteínas Repressoras/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (HNF1B protein, human); 0 (Heterochromatin); 0 (Histones); 0 (MIRN125 microRNA, human); 0 (MicroRNAs); 0 (Mirn125 microRNA, mouse); 0 (Repressor Proteins); 138674-15-4 (Hepatocyte Nuclear Factor 1-beta); EC 2.1.1. (SUV39H1 protein, human); EC 2.1.1.- (Methyltransferases); K3Z4F929H6 (Lysine)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171009
[Lr] Data última revisão:
171009
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170811
[St] Status:MEDLINE
[do] DOI:10.1074/jbc.M117.796771


  5 / 3299 MEDLINE  
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[PMID]:28665977
[Au] Autor:Wang P; Li X; Cao L; Huang S; Li H; Zhang Y; Yang T; Jiang J; Shi D
[Ad] Endereço:State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning, Guangxi Zhuang Autonomous Region, China.
[Ti] Título:MicroRNA-148a overexpression improves the early development of porcine somatic cell nuclear transfer embryos.
[So] Source:PLoS One;12(6):e0180535, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Incomplete epigenetic reprogramming of donor cell nuclei is one of the main contributors to the low efficiency of somatic cell nuclear transfer (SCNT). To improve the success of SCNT, somatic cell DNA methylation levels must be reduced to those levels found in totipotent embryonic cells. Recent studies have demonstrated that miR-148a can affect DNA methylation via DNMT1 modulation in various cancers. Therefore, the focus of this study was to examine the influence of miR-148a on DNA methylation in donor cells and in SCNT embryo development. Thus, a stable cell line overexpressing miR-148a was established and used to produce SCNT embryos. Upon examination, DNMT1 was found to be a miR-148a target in porcine fetal fibroblasts (PFF). Furthermore, miR-148a overexpression in PFFs significantly decreased DNMT1 expression and global DNA methylation levels (P < 0.05). Moreover, miRNA-148a expression levels in SCNT embryos were significantly lower at the 2-cell and 4-cell stages when compared to IVF and parthenogenetic embryos. The group overexpressing miRNA-148a also showed a significant increase in blastocyst formation and total cell numbers (P < 0.05). Additionally, miR-148a overexpression altered the immunofluorescence signal of 5-mC and H3K9ac, and enhanced pluripotent gene (Oct4 and Nanog) expression levels during embryo development. These results indicate that miR-148a overexpression enhances the developmental potential of SCNT embryos and modifies epigenetic status.
[Mh] Termos MeSH primário: Desenvolvimento Embrionário/genética
MicroRNAs/genética
Técnicas de Transferência Nuclear
Suínos/genética
[Mh] Termos MeSH secundário: Animais
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (MicroRNAs)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171006
[Lr] Data última revisão:
171006
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170701
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0180535


  6 / 3299 MEDLINE  
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[PMID]:28586343
[Au] Autor:Lee SE; Hyun H; Park MR; Choi Y; Son YJ; Park YG; Jeong SG; Shin MY; Ha HJ; Hong HS; Choi MK; Im GS; Park EW; Kim YH; Park C; Kim EY; Park SP
[Ad] Endereço:Faculty of Biotechnology, College of Applied Life Sciences, Jeju National University, Jeju-si, Jeju Special Self-Governing Province, Korea.
[Ti] Título:Production of transgenic pig as an Alzheimer's disease model using a multi-cistronic vector system.
[So] Source:PLoS One;12(6):e0177933, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Alzheimer's disease (AD) is a progressive neurodegenerative disease associated with memory loss and cognitive impairments. An AD transgenic (Tg) pig model would be useful for preclinical testing of therapeutic agents. We generated an AD Tg pig by somatic cell nuclear transfer (SCNT) using a multi-cistronic vector that harbored three AD-related genes with a total of six well-characterized mutations: hAPP (K670N/M671L, I716V, and V717I), hTau (P301L), and hPS1 (M146V and L286P). Four AD Tg cell lines were established from Jeju black pig ear fibroblasts (JB-PEFs); the resultant JB-PEFAD cells harbored transgene integration, expressed transgene mRNAs, and had normal karyotypes. Tg line #2-1, which expressed high levels of the transgenes, was used for SCNT; cleavage and blastocyst rates of embryos derived from this line were lower than those of Non-Tg. These embryos yielded three piglets (Jeju National University AD-Tg pigs, JNUPIGs) revealed by microsatellite testing to be genetically identical to JB-PEFAD. Transgenes were expressed in multiple tissues, and at especially high levels in brain, and Aß-40/42, total Tau, and GFAP levels were high in brains of the Tg animals. Five or more copies of transgenes were inserted into chromosome X. This is the first report of an AD Tg pig derived from a multi-cistronic vector.
[Mh] Termos MeSH primário: Doença de Alzheimer/genética
Animais Geneticamente Modificados/genética
Técnicas de Transferência Nuclear
Transgenes/genética
[Mh] Termos MeSH secundário: Doença de Alzheimer/tratamento farmacológico
Doença de Alzheimer/patologia
Animais
Blastocisto/metabolismo
Linhagem Celular
Modelos Animais de Doenças
Fibroblastos/metabolismo
Fibroblastos/patologia
Vetores Genéticos
Seres Humanos
Mutação
Suínos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170914
[Lr] Data última revisão:
170914
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170607
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0177933


  7 / 3299 MEDLINE  
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[PMID]:28576977
[Au] Autor:Clancy-Thompson E; Chen GZ; Tyler PM; Servos MM; Barisa M; Brennan PJ; Ploegh HL; Dougan SK
[Ad] Endereço:Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, MA 02215.
[Ti] Título:Monoclonal Invariant NKT (iNKT) Cell Mice Reveal a Role for Both Tissue of Origin and the TCR in Development of iNKT Functional Subsets.
[So] Source:J Immunol;199(1):159-171, 2017 Jul 01.
[Is] ISSN:1550-6606
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Invariant NKT (iNKT) cell functional subsets are defined by key transcription factors and output of cytokines, such as IL-4, IFN-γ, IL-17, and IL-10. To examine how TCR specificity determines iNKT function, we used somatic cell nuclear transfer to generate three lines of mice cloned from iNKT nuclei. Each line uses the invariant Vα14Jα18 TCRα paired with unique Vß7 or Vß8.2 subunits. We examined tissue homing, expression of PLZF, T-bet, and RORγt, and cytokine profiles and found that, although monoclonal iNKT cells differentiated into all functional subsets, the NKT17 lineage was reduced or expanded depending on the TCR expressed. We examined iNKT thymic development in limited-dilution bone marrow chimeras and show that higher TCR avidity correlates with higher PLZF and reduced T-bet expression. iNKT functional subsets showed distinct tissue distribution patterns. Although each individual monoclonal TCR showed an inherent subset distribution preference that was evident across all tissues examined, the iNKT cytokine profile differed more by tissue of origin than by TCR specificity.
[Mh] Termos MeSH primário: Diferenciação Celular
Células T Matadoras Naturais/imunologia
Receptores de Antígenos de Linfócitos T alfa-beta/metabolismo
Subpopulações de Linfócitos T/imunologia
[Mh] Termos MeSH secundário: Animais
Linfócitos B/imunologia
Linfócitos B/fisiologia
Citocinas/genética
Citocinas/imunologia
Citotoxicidade Imunológica/imunologia
Interleucina-10/imunologia
Interleucina-10/metabolismo
Interleucina-17/imunologia
Interleucina-17/metabolismo
Fatores de Transcrição Kruppel-Like/genética
Camundongos
Camundongos Endogâmicos C57BL
Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/genética
Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/imunologia
Técnicas de Transferência Nuclear
Especificidade de Órgãos
Proteína com Dedos de Zinco da Leucemia Promielocítica
Receptores de Antígenos de Linfócitos T alfa-beta/genética
Receptores de Antígenos de Linfócitos T alfa-beta/imunologia
Proteínas com Domínio T-Box/genética
Proteínas com Domínio T-Box/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Cytokines); 0 (Interleukin-17); 0 (Kruppel-Like Transcription Factors); 0 (Nuclear Receptor Subfamily 1, Group F, Member 3); 0 (Promyelocytic Leukemia Zinc Finger Protein); 0 (Receptors, Antigen, T-Cell, alpha-beta); 0 (T-Box Domain Proteins); 0 (T-box transcription factor TBX21); 0 (Zbtb16 protein, mouse); 130068-27-8 (Interleukin-10)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:171116
[Lr] Data última revisão:
171116
[Sb] Subgrupo de revista:AIM; IM
[Da] Data de entrada para processamento:170604
[St] Status:MEDLINE
[do] DOI:10.4049/jimmunol.1700214


  8 / 3299 MEDLINE  
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[PMID]:28545049
[Au] Autor:Wani NA; Vettical BS; Hong SB
[Ad] Endereço:Reproductive Biotechnology Center, Dubai, UAE.
[Ti] Título:First cloned Bactrian camel (Camelus bactrianus) calf produced by interspecies somatic cell nuclear transfer: A step towards preserving the critically endangered wild Bactrian camels.
[So] Source:PLoS One;12(5):e0177800, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Studies were conducted to explore the possibility of employing dromedary camel (Camelus dromedarius) oocytes as recipient cytoplasts for the development of interspecies somatic cell nuclear transfer (iSCNT) embryos using skin fibroblast cells of an adult Bactrian camel (Camelus bactrianus) and Llama (Llama glama) as donor nuclei. Also, the embryos reconstructed with Bactrian cells were transferred into the uterus of synchronized dromedary camel recipients to explore the possibility of using them as surrogate mothers. Serum-starved skin fibroblast cells were injected into the perivitelline space of enucleated mature oocytes, collected from super-stimulated dromedary camels, and fused using an Eppendorf electroporator. After activation with 5µM ionomycin and 6-dimethylaminopurine, they were cultured at 38.5°C in an atmosphere of 5% CO2, 5% O2, and 90% N2 in air. In experiment 1, Day 7 blastocysts were stained with Hoechst to count their cell numbers, while in experiment 2, they were transferred to synchronized dromedary recipients. A lower number (P < 0.05) of blastocysts were obtained from reconstructs utilizing fibroblast cells from Llama when compared with those reconstructed with dromedary and Bactrian fibroblast cells. However, no difference was observed in their cell numbers. In experiment 2, a higher (P < 0.05) proportion of blastocysts were obtained from the cleaved embryos reconstructed with Bactrian fibroblast cells when compared to those reconstructed with dromedary cells. Twenty-six Day 7 blastocysts reconstructed with Bactrian cells were transferred to 23 synchronized dromedary recipients with 5 pregnancies established on Day 30, however, only one of the pregnancies developed to term and a healthy calf weighing 33 kgs was born after completing 392 days of gestation. Unfortunately, the calf died on day 7 due to acute septicemia. In conclusion, the present study reports, for the first time, birth of a cloned Bactrian calf by iSCNT using dromedary camel as a source for oocytes as well as a surrogate for carrying the pregnancy to term.
[Mh] Termos MeSH primário: Clonagem de Organismos/veterinária
Transferência Embrionária/veterinária
Técnicas de Transferência Nuclear/veterinária
Indução da Ovulação/veterinária
[Mh] Termos MeSH secundário: Animais
Animais Recém-Nascidos
Camelídeos Americanos
Camelus
Técnicas de Cultura Embrionária
Espécies em Perigo de Extinção
Feminino
Gravidez
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170919
[Lr] Data última revisão:
170919
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170526
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0177800


  9 / 3299 MEDLINE  
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[PMID]:28407864
[Au] Autor:Kuwayama H; Tanabe Y; Wakayama T; Kishigami S
[Ad] Endereço:Faculty of Life and Environmental Sciences, University of Yamanashi, Yamanashi, Japan.
[Ti] Título:Birth of cloned mice from vaginal smear cells after somatic cell nuclear transfer.
[So] Source:Theriogenology;94:79-85, 2017 May.
[Is] ISSN:1879-3231
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Less invasive methods for donor cell collection will facilitate reproduction of wild animals using somatic-cell nuclear transfer. Stages of the estrous cycle in mice have long been studies using somatic cells that can be collected from vaginal walls using cotton tipped swabs in a relatively non-invasive manner. In this study, we examined the feasibility of these cells as sources of nuclei for somatic-cell cloning using nuclear transfer. Estrous cycles generally comprise proestrus, estrus, metestrus, and diestrus stages. In the present experiments, more than 60% of cells were nucleated in vaginal smears from all but the estrus stage. However, after somatic-cell nuclear transfer of cells from proestrus, metestrus, and diestrus stages, 66%, 50%, and 72% of cloned embryos developed to the morula/blastocyst, and cloned female mouse birth rates after embryo transfer were 1.5%, 0.3%, and 1%, respectively. These results show that noninvasively collected vaginal smears contain somatic cells that can be used to clone female mice.
[Mh] Termos MeSH primário: Clonagem de Organismos
Camundongos/embriologia
Técnicas de Transferência Nuclear/veterinária
Esfregaço Vaginal/veterinária
[Mh] Termos MeSH secundário: Animais
Coeficiente de Natalidade
Transferência Embrionária/veterinária
Desenvolvimento Embrionário
Estro
Feminino
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171009
[Lr] Data última revisão:
171009
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170415
[St] Status:MEDLINE


  10 / 3299 MEDLINE  
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[PMID]:28407860
[Au] Autor:Shen K; Li X; Dai X; Wang P; Li S; Xiong Z; Chen P; Liu Q; Shi D
[Ad] Endereço:State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning 530005, PR China.
[Ti] Título:Effects of MG132 on the in vitro development and epigenetic modification of Debao porcine somatic cell nuclear transfer embryos.
[So] Source:Theriogenology;94:48-58, 2017 May.
[Is] ISSN:1879-3231
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The present study was undertaken to examine the effect of MG132, a proteasome inhibitor, on the in vitro development, zygotic genome activation (ZGA) and epigenetic modification of Debao porcine somatic cell nuclear transfer (SCNT) embryos. Treatment of oocytes with 1 µM MG132 from 30 h to 42 h of maturation and SCNT embryos with 5 µM MG132 for 2 h after fusion resulted in higher blastocyst yield (36.5%) of SCNT embryos compared with the control group (11.0%). The ZGA of SCNT embryos at 2- and 4-cell stages was also enhanced by MG132 treatment through altering the RNA pol II status and increasing the expression of eIF3A and TFIIA. Meanwhile, MG132 treatment also resulted in increase of inner cell mass (ICM) and trophectoderm (TE) and total cell numbers and decrease of apoptotic cell numbers of SCNT blastocysts. Expression of BCL-2, OCT4, NANOG and CDX2 in SCNT blastocysts developed from SCNT embryos and oocytes treated with MG132 was increased significantly (P < 0.01), while the expression of pro-apoptotic BAX gene was suppressed significantly (P < 0.01). In addition, MG132 treatment not only affected the expression patterns of H3K9 acetylation, H3K4 and H3K9 trimethylation, but also regulated the relative expression of SMYD3, ASH2L, KDM5B, HAT1, HDAC1 and HDAC2 of Debao porcine SCNT embryos. These results demonstrate that MG132 treatment can improve the developmental potential of Debao porcine SCNT embryos through regulating the expression of genes related to histone acetylation and the processes of ZGA.
[Mh] Termos MeSH primário: Desenvolvimento Embrionário/efeitos dos fármacos
Leupeptinas/farmacologia
Inibidores de Proteassoma/farmacologia
Suínos/embriologia
[Mh] Termos MeSH secundário: Acetilação/efeitos dos fármacos
Animais
Apoptose/efeitos dos fármacos
Blastocisto/efeitos dos fármacos
Massa Celular Interna do Blastocisto/efeitos dos fármacos
Técnicas de Cultura Embrionária/veterinária
Epigênese Genética
Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos
Histonas/metabolismo
Técnicas de Transferência Nuclear/veterinária
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Histones); 0 (Leupeptins); 0 (Proteasome Inhibitors); RF1P63GW3K (benzyloxycarbonylleucyl-leucyl-leucine aldehyde)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171009
[Lr] Data última revisão:
171009
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170415
[St] Status:MEDLINE



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