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1.
Methods Mol Biol ; 2214: 265-282, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32944916

RESUMO

Investigating the chromatin landscape of the early mammalian embryo is essential to understand how epigenetic mechanisms may direct reprogramming and cell fate allocation. Genome-wide analyses of the epigenome in preimplantation mouse embryos have recently become available, thanks to the development of low-input protocols. DNA adenine methyltransferase identification (DamID) enables the investigation of genome-wide protein-DNA interactions without the requirement of specific antibodies. Most importantly, DamID can be robustly applied to single cells. Here we describe the protocol for performing DamID in single oocytes and mouse preimplantation embryos, as well as single blastomeres, using a Dam-LaminB1 fusion to generate high-resolution lamina-associated domain (LAD) maps. This low-input method can be adapted for other proteins of interest to faithfully profile their genomic interaction, allowing us to interrogate the chromatin dynamics and nuclear organization during the early mammalian development.


Assuntos
Blastocisto/metabolismo , Genômica/métodos , Camundongos/embriologia , Camundongos/genética , DNA Metiltransferases Sítio Específica (Adenina-Específica)/metabolismo , Animais , Blastocisto/citologia , Células Cultivadas , Técnicas de Cultura Embrionária/métodos , Genoma , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Camundongos/metabolismo , Camundongos Endogâmicos C57BL , Oócitos/citologia , Oócitos/metabolismo , Análise de Célula Única/métodos
2.
Nucleic Acids Res ; 49(2): 657-673, 2021 01 25.
Artigo em Inglês | MEDLINE | ID: mdl-33367834

RESUMO

Antisense oligonucleotides (ASOs) have emerged as a new class of drugs to treat a wide range of diseases, including neurological indications. Spinraza, an ASO that modulates splicing of SMN2 RNA, has shown profound disease modifying effects in Spinal Muscular Atrophy (SMA) patients, energizing efforts to develop ASOs for other neurological diseases. While SMA specifically affects spinal motor neurons, other neurological diseases affect different central nervous system (CNS) regions, neuronal and non-neuronal cells. Therefore, it is important to characterize ASO distribution and activity in all major CNS structures and cell types to have a better understanding of which neurological diseases are amenable to ASO therapy. Here we present for the first time the atlas of ASO distribution and activity in the CNS of mice, rats, and non-human primates (NHP), species commonly used in preclinical therapeutic development. Following central administration of an ASO to rodents, we observe widespread distribution and target RNA reduction throughout the CNS in neurons, oligodendrocytes, astrocytes and microglia. This is also the case in NHP, despite a larger CNS volume and more complex neuroarchitecture. Our results demonstrate that ASO drugs are well suited for treating a wide range of neurological diseases for which no effective treatments are available.


Assuntos
Sistema Nervoso Central/química , Camundongos/metabolismo , Oligonucleotídeos Antissenso/farmacocinética , Primatas/metabolismo , Ratos/metabolismo , Animais , Sistema Nervoso Central/citologia , Feminino , Hibridização In Situ , Injeções Intraventriculares , Injeções Espinhais , Macaca fascicularis , Masculino , Neuroglia/química , Neurônios/química , Oligonucleotídeos Antissenso/administração & dosagem , Especificidade de Órgãos , RNA Longo não Codificante/análise , RNA Longo não Codificante/antagonistas & inibidores , RNA Longo não Codificante/genética , Ratos Sprague-Dawley , Ribonuclease H , Distribuição Tecidual
3.
Proc Natl Acad Sci U S A ; 117(33): 20015-20026, 2020 08 18.
Artigo em Inglês | MEDLINE | ID: mdl-32759216

RESUMO

We sequenced more than 52,500 single cells from embryonic day 11.5 (E11.5) postembryonic day 5 (P5) gonads and performed lineage tracing to analyze primordial follicles and wave 1 medullar follicles during mouse fetal and perinatal oogenesis. Germ cells clustered into six meiotic substages, as well as dying/nurse cells. Wnt-expressing bipotential precursors already present at E11.5 are followed at each developmental stage by two groups of ovarian pregranulosa (PG) cells. One PG group, bipotential pregranulosa (BPG) cells, derives directly from bipotential precursors, expresses Foxl2 early, and associates with cysts throughout the ovary by E12.5. A second PG group, epithelial pregranulosa (EPG) cells, arises in the ovarian surface epithelium, ingresses cortically by E12.5 or earlier, expresses Lgr5, but delays robust Foxl2 expression until after birth. By E19.5, EPG cells predominate in the cortex and differentiate into granulosa cells of quiescent primordial follicles. In contrast, medullar BPG cells differentiate along a distinct pathway to become wave 1 granulosa cells. Reflecting their separate somatic cellular lineages, second wave follicles were ablated by diptheria toxin treatment of Lgr5-DTR-EGFP mice at E16.5 while first wave follicles developed normally and supported fertility. These studies provide insights into ovarian somatic cells and a resource to study the development, physiology, and evolutionary conservation of mammalian ovarian follicles.


Assuntos
Células da Granulosa/citologia , Camundongos/embriologia , Folículo Ovariano/embriologia , Animais , Diferenciação Celular , Linhagem da Célula , Feminino , Proteína Forkhead Box L2/genética , Proteína Forkhead Box L2/metabolismo , Células da Granulosa/metabolismo , Camundongos/metabolismo , Folículo Ovariano/citologia , Folículo Ovariano/metabolismo , Gravidez , Receptores Acoplados a Proteínas-G/genética , Receptores Acoplados a Proteínas-G/metabolismo
4.
Nat Commun ; 11(1): 3603, 2020 07 17.
Artigo em Inglês | MEDLINE | ID: mdl-32681107

RESUMO

Members of the PR/SET domain-containing (PRDM) family of zinc finger transcriptional regulators play diverse developmental roles. PRDM10 is a yet uncharacterized family member, and its function in vivo is unknown. Here, we report an essential requirement for PRDM10 in pre-implantation embryos and embryonic stem cells (mESCs), where loss of PRDM10 results in severe cell growth inhibition. Detailed genomic and biochemical analyses reveal that PRDM10 functions as a sequence-specific transcription factor. We identify Eif3b, which encodes a core component of the eukaryotic translation initiation factor 3 (eIF3) complex, as a key downstream target, and demonstrate that growth inhibition in PRDM10-deficient mESCs is in part mediated through EIF3B-dependent effects on global translation. Our work elucidates the molecular function of PRDM10 in maintaining global translation, establishes its essential role in early embryonic development and mESC homeostasis, and offers insights into the functional repertoire of PRDMs as well as the transcriptional mechanisms regulating translation.


Assuntos
Regulação da Expressão Gênica no Desenvolvimento , Camundongos/metabolismo , Fatores de Transcrição/metabolismo , Animais , Desenvolvimento Embrionário , Células-Tronco Embrionárias/metabolismo , Fatores de Iniciação em Eucariotos/genética , Fatores de Iniciação em Eucariotos/metabolismo , Feminino , Peptídeos e Proteínas de Sinalização Intracelular/genética , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Masculino , Camundongos/embriologia , Camundongos/genética , Biossíntese de Proteínas , Fatores de Transcrição/genética
5.
Nat Commun ; 11(1): 635, 2020 01 31.
Artigo em Inglês | MEDLINE | ID: mdl-32005814

RESUMO

Multipotent Nkx2-1-positive lung epithelial primordial progenitors of the foregut endoderm are thought to be the developmental precursors to all adult lung epithelial lineages. However, little is known about the global transcriptomic programs or gene networks that regulate these gateway progenitors in vivo. Here we use bulk RNA-sequencing to describe the unique genetic program of in vivo murine lung primordial progenitors and computationally identify signaling pathways, such as Wnt and Tgf-ß superfamily pathways, that are involved in their cell-fate determination from pre-specified embryonic foregut. We integrate this information in computational models to generate in vitro engineered lung primordial progenitors from mouse pluripotent stem cells, improving the fidelity of the resulting cells through unbiased, easy-to-interpret similarity scores and modulation of cell culture conditions, including substratum elastic modulus and extracellular matrix composition. The methodology proposed here can have wide applicability to the in vitro derivation of bona fide tissue progenitors of all germ layers.


Assuntos
Células Epiteliais/citologia , Pulmão/citologia , Camundongos/genética , Células-Tronco Pluripotentes/citologia , Animais , Técnicas de Cultura de Células , Diferenciação Celular , Células Epiteliais/metabolismo , Matriz Extracelular/genética , Matriz Extracelular/metabolismo , Feminino , Camadas Germinativas/embriologia , Camadas Germinativas/metabolismo , Pulmão/embriologia , Pulmão/metabolismo , Masculino , Camundongos/embriologia , Camundongos/metabolismo , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Células-Tronco Pluripotentes/metabolismo , Transdução de Sinais , Fator Nuclear 1 de Tireoide/genética , Fator Nuclear 1 de Tireoide/metabolismo , Transcriptoma , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta/metabolismo
6.
J Endocrinol ; 244(2): 273-283, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31671405

RESUMO

Kiss1 neurons of the arcuate (ARC) nucleus form an interconnected network of cells that communicate via neurokinin B (encoded by Tac2) and its receptor (encoded by Tacr3) and play key roles in the control of the reproductive axis through sex hormone-regulated synthesis and release of kisspeptin peptides (Kp, encoded by Kiss1). The aim of this study was to determine whether the Kiss1 cell population of the ARC already displays sexually dimorphic features at embryonic age E16.5 in mice. At this time of development, Kiss1-GFP- and Kp-immunoreactive cell bodies were restricted to the ARC and not found in the pre-optic area (POA). The Kiss1-GFP cell population was identical in size between sexes but had significantly lower Kiss1, Tac2, and Tacr3 mRNA levels and lower Kp-ir fiber density in the POA in male compared to female fetuses. Receptors for androgen (Ar) and estrogen (Esr1, Esr2, Gpr30) and the Cyp19a1 gene (encoding the estradiol-producing enzyme aromatase) transcripts were also detected in fetal ARC Kiss1-GFP cells with significant sex differences for Ar (higher in males) and Esr1 (higher in females). Functional studies on primary cultures of sorted fetal Kiss1-GFP cells revealed a significant negative effect of estradiol treatment on neurite outgrowth on the fourth day of culture in the female group specifically. We conclude that the ARC Kiss1 cell population is already sexually differentiated at E16.5 and that its morphogenetic development may be particularly vulnerable to estradiol exposure at this early developmental time.


Assuntos
Estradiol/metabolismo , Kisspeptinas/metabolismo , Camundongos/metabolismo , Neuritos/metabolismo , Caracteres Sexuais , Animais , Núcleo Arqueado do Hipotálamo/metabolismo , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Kisspeptinas/genética , Masculino , Camundongos/embriologia , Camundongos/genética , Camundongos Endogâmicos C57BL , Neurocinina B/genética
7.
Int J Environ Health Res ; 30(2): 117-133, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30758226

RESUMO

The relationship between air pollution exposure and haematology remains controversial. Evidences in the effect of trace organic air pollutants and in the impact of such exposure on lipid and protein levels are scarce. This work investigated the health effects of medium-term exposure to traffic-related air pollution on both haematological and biochemical indices in animal models. Two groups of albino mice (Mus musculus) were exposed to ambient air polluted by vehicle exhaust for three and six months, and one group was kept as control. Results found significant depletions (p < 0.05) in red blood cells, packed cell volume, neutrophils, eosinophils, monocytes, and total cholesterol after air pollution exposure. On the contrary, significant elevations (p < 0.05) were observed in platelet, lymphocytes, and serum albumin compared to control condition. Correlation data suggested that significant changes in blood parameters may be altered by the synergistic effect of several organic and inorganic air pollutants.


Assuntos
Poluentes Atmosféricos/análise , Exposição Ambiental/análise , Camundongos/metabolismo , Emissões de Veículos/análise , Poluição do Ar/análise , Animais , Análise Química do Sangue , Feminino , Testes Hematológicos , Masculino , Camundongos/sangue
8.
Nat Commun ; 10(1): 5176, 2019 11 15.
Artigo em Inglês | MEDLINE | ID: mdl-31729371

RESUMO

Hematopoiesis in mammalian embryos proceeds through three successive waves of hematopoietic progenitors. Since their emergence spatially and temporally overlap and phenotypic markers are often shared, the specifics regarding their origin, development, lineage restriction and mutual relationships have not been fully determined. The identification of wave-specific markers would aid to resolve these uncertainties. Here, we show that toll-like receptors (TLRs) are expressed during early mouse embryogenesis. We provide phenotypic and functional evidence that the expression of TLR2 on E7.5 c-kit+ cells marks the emergence of precursors of erythro-myeloid progenitors (EMPs) and provides resolution for separate tracking of EMPs from primitive progenitors. Using in vivo fate mapping, we show that at E8.5 the Tlr2 locus is already active in emerging EMPs and in progenitors of adult hematopoietic stem cells (HSC). Together, this data demonstrates that the activation of the Tlr2 locus tracks the earliest events in the process of EMP and HSC specification.


Assuntos
Células-Tronco Hematopoéticas/metabolismo , Camundongos/embriologia , Proteínas Proto-Oncogênicas c-kit/metabolismo , Receptor 2 Toll-Like/metabolismo , Células-Tronco Adultas/metabolismo , Animais , Feminino , Hematopoese , Masculino , Camundongos/genética , Camundongos/metabolismo , Camundongos Endogâmicos C57BL , Proteínas Proto-Oncogênicas c-kit/genética , Receptor 2 Toll-Like/genética
9.
Comp Med ; 69(5): 350-373, 2019 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-31540584

RESUMO

Light is a potent biologic force that profoundly influences circadian, neuroendocrine, and neurobehavioral regulation in animals. Previously we examined the effects of light-phase exposure of rats to white light-emitting diodes (LED), which emit more light in the blue-appearing portion of the visible spectrum (465 to 485 nm) than do broad-spectrum cool white fluorescent (CWF) light, on the nighttime melatonin amplitude and circadian regulation of metabolism and physiology. In the current studies, we tested the hypothesis that exposure to blue-enriched LED light at day (bLAD), compared with CWF, promotes the circadian regulation of neuroendocrine, metabolic, and physiologic parameters that are associated with optimizing homeostatic regulation of health and wellbeing in 3 mouse strains commonly used in biomedical research (C3H [melatonin-producing], C57BL/6, and BALB/c [melatonin-non-producing]). Compared with male and female mice housed for 12 wk under 12:12-h light:dark (LD) cycles in CWF light, C3H mice in bLAD evinced 6-fold higher peak plasma melatonin levels at the middark phase; in addition, high melatonin levels were prolonged 2 to 3 h into the light phase. C57BL/6 and BALB/c strains did not produce nighttime pineal melatonin. Body growth rates; dietary and water intakes; circadian rhythms of arterial blood corticosterone, insulin, leptin, glucose, and lactic acid; pO2 and pCO2; fatty acids; and metabolic indicators (cAMP, DNA, tissue DNA 3H-thymidine incorporation, fat content) in major organ systems were significantly lower and activation of major metabolic signaling pathways (mTOR, GSK3ß, and SIRT1) in skeletal muscle and liver were higher only in C3H mice in bLAD compared with CWF. These data show that exposure of C3H mice to bLAD compared with CWF has a marked positive effect on the circadian regulation of neuroendocrine, metabolic, and physiologic parameters associated with the promotion of animal health and wellbeing that may influence scientific outcomes. The absence of enhancement in amelatonic strains suggests hyperproduction of nighttime melatonin may be a key component of the physiology.


Assuntos
Ritmo Circadiano/fisiologia , Luz , Camundongos Endogâmicos BALB C/metabolismo , Camundongos Endogâmicos C3H/metabolismo , Camundongos Endogâmicos C57BL/metabolismo , Animais , Feminino , Masculino , Melatonina/sangue , Camundongos/metabolismo
10.
J Agric Food Chem ; 67(30): 8303-8311, 2019 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-31298535

RESUMO

Exposure to chiral pesticides poses many potential health risks. In this study, we examined the impacts of exposure to penconazole and its enantiomers on gut microbiota and metabolic profiles in mice. The relative abundance of microbiota in cecal content significantly changed following exposure to penconazole and its enantiomers. At the genus level, the relative abundances of seven gut microflora were altered following exposure to (-)-penconazole. Both (±)-penconazole and (+)-penconazole caused significant changes in the relative abundances of five gut microflora. In addition, targeted serum metabolomics analysis showed disturbed metabolic profiles following exposure. Respectively, (±)-penconazole, (+)-penconazole, and (-)-penconazole exposure significantly altered the relative levels of 29, 23, and 36 metabolites. In general, exposure to penconazole and its enantiomers caused disorders in gut microbiota and metabolic profiles of mice. The potential health risks of penconazole and its enantiomers now require further evaluation.


Assuntos
Microbioma Gastrointestinal/efeitos dos fármacos , Camundongos/metabolismo , Praguicidas/química , Praguicidas/farmacologia , Triazóis/química , Triazóis/farmacologia , Animais , Bactérias/efeitos dos fármacos , Bactérias/genética , Bactérias/isolamento & purificação , Trato Gastrointestinal/microbiologia , Masculino , Camundongos Endogâmicos ICR , Filogenia , Estereoisomerismo
11.
Chem Biol Interact ; 310: 108715, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31226285

RESUMO

Although the three-dimensional structures of mouse and Torpedo californica acetylcholinesterase are very similar, their responses to the covalent sulfonylating agents benzenesulfonyl fluoride and phenylmethylsulfonyl fluoride are qualitatively different. Both agents inhibit the mouse enzyme effectively by covalent modification of its active-site serine. In contrast, whereas the Torpedo enzyme is effectively inhibited by benzenesulfonyl fluoride, it is almost completely resistant to phenylmethylsulfonyl fluoride. A bottleneck midway down the active-site gorge in both enzymes restricts access of ligands to the active site at the bottom of the gorge. Molecular dynamics simulations revealed that the mouse enzyme is substantially more flexible than the Torpedo enzyme, suggesting that enhanced 'breathing motions' of the mouse enzyme relative to the Torpedo enzyme may explain why phenylmethylsulfonyl fluoride can reach the active site in mouse acetylcholinesterase, but not in the Torpedo enzyme. Accordingly, we performed docking of the two sulfonylating agents to the two enzymes, followed by molecular dynamics simulations. Whereas benzenesulfonyl fluoride closely approaches the active-site serine in both mouse and Torpedo acetylcholinesterase in such simulations, phenylmethylsulfonyl fluoride is able to approach the active-site serine of mouse acetylcholinesterase, but remains trapped above the bottleneck in the Torpedo enzyme. Our studies demonstrate that reliance on docking tools in drug design can produce misleading information. Docking studies should, therefore, also be complemented by molecular dynamics simulations in selection of lead compounds. An animated Interactive 3D Complement (I3DC) is available in Proteopedia at http://proteopedia.org/w/Journal:CHEMBIOINT:2.


Assuntos
Acetilcolinesterase/metabolismo , Inibidores da Colinesterase/metabolismo , Desenho de Fármacos , Simulação de Dinâmica Molecular , Animais , Benzenossulfonatos/metabolismo , Domínio Catalítico , Fluoretos/metabolismo , Humanos , Camundongos/metabolismo , Simulação de Acoplamento Molecular , Fluoreto de Fenilmetilsulfonil/metabolismo , Especificidade da Espécie , Torpedo/metabolismo
12.
Nat Commun ; 10(1): 2835, 2019 06 27.
Artigo em Inglês | MEDLINE | ID: mdl-31249377

RESUMO

During embryogenesis cells make fate decisions within complex tissue environments. The levels and dynamics of transcription factor expression regulate these decisions. Here, we use single cell live imaging of an endogenous HES5 reporter and absolute protein quantification to gain a dynamic view of neurogenesis in the embryonic mammalian spinal cord. We report that dividing neural progenitors show both aperiodic and periodic HES5 protein fluctuations. Mathematical modelling suggests that in progenitor cells the HES5 oscillator operates close to its bifurcation boundary where stochastic conversions between dynamics are possible. HES5 expression becomes more frequently periodic as cells transition to differentiation which, coupled with an overall decline in HES5 expression, creates a transient period of oscillations with higher fold expression change. This increases the decoding capacity of HES5 oscillations and correlates with interneuron versus motor neuron cell fate. Thus, HES5 undergoes complex changes in gene expression dynamics as cells differentiate.


Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Células-Tronco Neurais/metabolismo , Neurogênese , Proteínas Repressoras/metabolismo , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/química , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Masculino , Camundongos/embriologia , Camundongos/metabolismo , Camundongos Endogâmicos ICR , Camundongos Knockout , Células-Tronco Neurais/química , Células-Tronco Neurais/citologia , Proteínas Repressoras/química , Proteínas Repressoras/genética , Análise de Célula Única
13.
Exp Anim ; 68(3): 267-275, 2019 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-30745527

RESUMO

Lipoprotein lipase (LPL) and hepatic triglyceride lipase (HTGL) have an important role in lifestyle-related diseases. To evaluate species differences, we compared LPL and HTGL activities in different animal models of lifestyle-related diseases using the same assay kit. Normal animals (JW rabbits, ICR mice, and SD rats), a hypercholesterolemic animal model (WHHLMI rabbits), and obese animal models (KK-Ay mice and Zucker fatty rats) fed standard chow were used in this study. Plasma was prepared before and after an intravenous injection of heparin sodium under fasting and feeding. LPL and HTGL activities were measured with the LPL/HTGL activity assay kit (Immuno-Biological Laboratories) using an auto-analyzer. Only in mice, high HTGL activity was observed in pre-heparin plasma. In normal animals, LPL and HTGL activities were high in ICR mice and SD rats but low in JW rabbits. Compared to normal animals, LPL activity was high in Zucker fatty rats and WHHLMI rabbits at both fasting and feeding, while LPL activity after feeding was low in KK-Ay mice. HTGL activity was higher in fasted and fed WHHLMI rabbits and fasted Zucker fatty rats, but was lower in fed KK-Ay mice. Gender difference was observed in HTGL activity in SD rats and LPL activity in WHHLMI rabbits but not in ICR mice. In conclusion, this simple assay method was effective for measuring LPL and HTGL activities of experimental animals, and the activities are highly regulated depending on animal species, animal models, feeding/fasting conditions and genders.


Assuntos
Ensaios Enzimáticos Clínicos/métodos , Lipase/sangue , Lipase Lipoproteica/sangue , Camundongos/metabolismo , Coelhos/metabolismo , Ratos/metabolismo , Animais , Modelos Animais de Doenças , Jejum , Feminino , Humanos , Masculino , Camundongos Endogâmicos ICR , Camundongos Obesos , Ratos Sprague-Dawley , Ratos Zucker , Especificidade da Espécie
14.
Medisan ; 23(1)ene.-feb. 2019. tab, graf
Artigo em Espanhol | LILACS | ID: biblio-990175

RESUMO

Se realizó un estudio prospectivo y experimental, a partir de la inducción de un trauma isquémico cerebral permanente, mediante sección de la carótida derecha, en 24 gerbils de Mongolia, procedentes del Centro Nacional para la Producción de Animales de Laboratorio, con vistas a identificar los posibles cambios en las poblaciones de neuronas de la fascia dentada del hipocampo durante la isquemia. Los resultados revelaron que la isquemia cerebral permanente produce destrucción progresiva y total de las neuronas en la fascia dentada durante las primeras 72 horas del proceso isquémico y muestra 3 momentos durante su evolución con características específicas para cada uno, así como cambios en las proporciones entre las neuronas en proceso de muerte celular, las aparentemente sanas y aquellas con diferentes cantidades de nucléolos en la zona dañada por el proceso isquémico.


A prospective and experimental study was carried out, from the induction of a permanent cerebral ischemic trauma, by means of a cut to the right carotid vein, in 24 gerbils from Mongolia, obtained from the National Center for Laboratory Animals Production, with the aim of identifying the possible changes in the neurons populations of the hypocampus dentate fascia during ischemia. The results revealed that the permanent cerebral ischemia produces progressive and total destruction of the neurons in the dentate fascia during the first 72 hours of the ischemic process and it shows 3 moments during its course, each of them with specific characteristics, as well as changes in the proportions among the neurons during cellular death, those apparently healthy and those with different quantities of nucleolous in the area damaged by the ischemic process.


Assuntos
Animais , Masculino , Camundongos , Infarto Cerebral , Isquemia Encefálica/líquido cefalorraquidiano , Gerbillinae/metabolismo , Camundongos/metabolismo , Epidemiologia Experimental , Estudos Prospectivos , Acidente Vascular Cerebral
15.
Medisan ; 23(1)ene.-feb. 2019. tab, graf
Artigo em Espanhol | CUMED | ID: cum-74721

RESUMO

Se realizó un estudio prospectivo y experimental, a partir de la inducción de un trauma isquémico cerebral permanente, mediante sección de la carótida derecha, en 24 gerbils de Mongolia, procedentes del Centro Nacional para la Producción de Animales de Laboratorio, con vistas a identificar los posibles cambios en las poblaciones de neuronas de la fascia dentada del hipocampo durante la isquemia. Los resultados revelaron que la isquemia cerebral permanente produce destrucción progresiva y total de las neuronas en la fascia dentada durante las primeras 72 horas del proceso isquémico y muestra 3 momentos durante su evolución con características específicas para cada uno, así como cambios en las proporciones entre las neuronas en proceso de muerte celular, las aparentemente sanas y aquellas con diferentes cantidades de nucléolos en la zona dañada por el proceso isquémico(AU)


A prospective and experimental study was carried out, from the induction of a permanent cerebral ischemic trauma, by means of a cut to the right carotid vein, in 24 gerbils from Mongolia, obtained from the National Center for Laboratory Animals Production, with the aim of identifying the possible changes in the neurons populations of the hypocampus dentate fascia during ischemia. The results revealed that the permanent cerebral ischemia produces progressive and total destruction of the neurons in the dentate fascia during the first 72 hours of the ischemic process and it shows 3 moments during its course, each of them with specific characteristics, as well as changes in the proportions among the neurons during cellular death, those apparently healthy and those with different quantities of nucleolous in the area damaged by the ischemic process(AU)


Assuntos
Animais , Infarto Cerebral , Isquemia Encefálica/líquido cefalorraquidiano , Gerbillinae/metabolismo , Camundongos/metabolismo , Epidemiologia Experimental , Estudos Prospectivos , Acidente Vascular Cerebral
16.
Mol Cell Biol ; 39(6)2019 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-30642949

RESUMO

The physiological functions of the atypical mitogen-activated protein kinase extracellular signal-regulated kinase 3 (ERK3) remain poorly characterized. Previous analysis of mice with a targeted insertion of the lacZ reporter in the Mapk6 locus (Mapk6lacZ ) showed that inactivation of ERK3 in Mapk6lacZ mice leads to perinatal lethality associated with intrauterine growth restriction, defective lung maturation, and neuromuscular anomalies. To further explore the role of ERK3 in physiology and disease, we generated novel mouse models expressing a catalytically inactive (Mapk6KD ) or conditional (Mapk6Δ ) allele of ERK3. Surprisingly, we found that mice devoid of ERK3 kinase activity or expression survive the perinatal period without any observable lung or neuromuscular phenotype. ERK3 mutant mice reached adulthood, were fertile, and showed no apparent health problem. However, analysis of growth curves revealed that ERK3 kinase activity is necessary for optimal postnatal growth. To gain insight into the genetic basis underlying the discrepancy in phenotypes of different Mapk6 mutant mouse models, we analyzed the regulation of genes flanking the Mapk6 locus by quantitative PCR. We found that the expression of several Mapk6 neighboring genes is deregulated in Mapk6lacZ mice but not in Mapk6KD or Mapk6Δ mutant mice. Our genetic analysis suggests that off-target effects of the targeting construct on local gene expression are responsible for the perinatal lethality phenotype of Mapk6lacZ mutant mice.


Assuntos
Camundongos/crescimento & desenvolvimento , Proteína Quinase 6 Ativada por Mitógeno/metabolismo , Animais , Modelos Animais de Doenças , Embrião de Mamíferos/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Sistema de Sinalização das MAP Quinases , Camundongos/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo
17.
Nat Commun ; 9(1): 5279, 2018 12 11.
Artigo em Inglês | MEDLINE | ID: mdl-30538245

RESUMO

Regulatory mechanisms controlling the pool size of spleen dendritic cells (DC) remain incompletely understood. DCs are continuously replenished from hematopoietic stem cells, and FLT3-mediated signals cell-intrinsically regulate homeostatic expansion of spleen DCs. Here we show that combining FLT3 and CSF1R-deficiencies results in specific and complete abrogation of spleen DCs in vivo. Spatiotemporally controlled CSF1R depletion reveals a cell-extrinsic and non-hematopoietic mechanism for DC pool size regulation. Lack of CSF1R-mediated signals impedes the differentiation of spleen macrophages of embryonic origin, and the resulted macrophage depletion during development or in adult mice results in loss of DCs. Moreover, embryo-derived macrophages are important for the physiologic regeneration of DC after activation-induced depletion in situ. In summary, we show that the differentiation of DC and their regeneration relies on ontogenetically distinct spleen macrophages, thereby providing a novel regulatory principle that may also be important for the differentiation of other hematopoietic cell types.


Assuntos
Células Dendríticas/citologia , Macrófagos/citologia , Camundongos/embriologia , Receptores de Fator Estimulador das Colônias de Granulócitos e Macrófagos/metabolismo , Animais , Diferenciação Celular , Células Dendríticas/metabolismo , Feminino , Macrófagos/metabolismo , Masculino , Camundongos/metabolismo , Camundongos Knockout , Receptores de Fator Estimulador das Colônias de Granulócitos e Macrófagos/genética , Baço/citologia , Baço/metabolismo , Tirosina Quinase 3 Semelhante a fms/genética , Tirosina Quinase 3 Semelhante a fms/metabolismo
19.
Artigo em Inglês | MEDLINE | ID: mdl-30397099

RESUMO

In eukaryotes, almost all RNA species are processed at their 3' ends and most mRNAs are polyadenylated in the nucleus by canonical poly(A) polymerases. In recent years, several terminal nucleotidyl transferases (TENTs) including non-canonical poly(A) polymerases (ncPAPs) and terminal uridyl transferases (TUTases) have been discovered. In contrast to canonical polymerases, TENTs' functions are more diverse; some, especially TUTases, induce RNA decay while others, such as cytoplasmic ncPAPs, activate translationally dormant deadenylated mRNAs. The mammalian genome encodes 11 different TENTs. This review summarizes the current knowledge about the functions and mechanisms of action of these enzymes.This article is part of the theme issue '5' and 3' modifications controlling RNA degradation'.


Assuntos
Camundongos/genética , Nucleotidiltransferases/genética , RNA/metabolismo , Ratos/genética , Animais , Camundongos/metabolismo , Nucleotidiltransferases/metabolismo , Estabilidade de RNA , Ratos/metabolismo
20.
Neuromolecular Med ; 20(4): 419-436, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30276585

RESUMO

Toll receptors, first identified to regulate embryogenesis and immune responses in the adult fly and subsequently defined as the principal sensors of infection in mammals, are increasingly appreciated for their impact on the homeostasis of the central as well as the peripheral nervous systems. Whereas in the context of immunity, the fly Toll and the mammalian TLR pathways have been researched in parallel, the expression pattern and functionality have largely been researched disparately. Herein, we provide data on the expression pattern of the Toll homologues, signaling components, and downstream effectors in ten different cell populations of the adult fly central nervous system (CNS). We have compared the expression of the different Toll pathways in the fly to the expression of TLRs in the mouse brain and discussed the implications with respect to commonalities, differences, and future perspectives.


Assuntos
Sistema Nervoso Central/metabolismo , Proteínas de Drosophila/fisiologia , Drosophila melanogaster/metabolismo , Camundongos/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Receptores Toll-Like/fisiologia , Animais , Encéfalo/metabolismo , Sistema Nervoso Central/imunologia , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Drosophila melanogaster/imunologia , Regulação da Expressão Gênica , Camundongos/imunologia , Neuroglia/metabolismo , Neurônios/metabolismo , Padrões Moleculares Associados a Patógenos/metabolismo , RNA Mensageiro/biossíntese , Transdução de Sinais , Especificidade da Espécie , Receptores Toll-Like/genética
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