Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 7 de 7
Filtrar
1.
Mol Cell ; 81(15): 3065-3081.e12, 2021 08 05.
Artículo en Inglés | MEDLINE | ID: mdl-34297911

RESUMEN

The chromatin fiber folds into loops, but the mechanisms controlling loop extrusion are still poorly understood. Using super-resolution microscopy, we visualize that loops in intact nuclei are formed by a scaffold of cohesin complexes from which the DNA protrudes. RNA polymerase II decorates the top of the loops and is physically segregated from cohesin. Augmented looping upon increased loading of cohesin on chromosomes causes disruption of Lamin at the nuclear rim and chromatin blending, a homogeneous distribution of chromatin within the nucleus. Altering supercoiling via either transcription or topoisomerase inhibition counteracts chromatin blending, increases chromatin condensation, disrupts loop formation, and leads to altered cohesin distribution and mobility on chromatin. Overall, negative supercoiling generated by transcription is an important regulator of loop formation in vivo.


Asunto(s)
Proteínas de Ciclo Celular/metabolismo , Cromatina/química , Cromatina/genética , Proteínas Cromosómicas no Histona/metabolismo , Transcripción Genética/fisiología , Proteínas de Ciclo Celular/química , Proteínas de Ciclo Celular/genética , Línea Celular , Núcleo Celular/genética , Proteoglicanos Tipo Condroitín Sulfato/genética , Proteoglicanos Tipo Condroitín Sulfato/metabolismo , Cromatina/metabolismo , Proteínas Cromosómicas no Histona/química , Proteínas Cromosómicas no Histona/genética , ADN-Topoisomerasas de Tipo I/genética , ADN-Topoisomerasas de Tipo I/metabolismo , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Femenino , Humanos , Laminas/genética , Laminas/metabolismo , ARN Polimerasa II/metabolismo , Imagen Individual de Molécula/métodos , Cohesinas
2.
Mol Ther ; 29(2): 804-821, 2021 02 03.
Artículo en Inglés | MEDLINE | ID: mdl-33264643

RESUMEN

Cell therapy approaches hold great potential for treating retinopathies, which are currently incurable. This study addresses the problem of inadequate migration and integration of transplanted cells into the host retina. To this end, we have identified the chemokines that were most upregulated during retinal degeneration and that could chemoattract mesenchymal stem cells (MSCs). The results were observed using a pharmacological model of ganglion/amacrine cell degeneration and a genetic model of retinitis pigmentosa, from both mice and human retinae. Remarkably, MSCs overexpressing Ccr5 and Cxcr6, which are receptors bound by a subset of the identified chemokines, displayed improved migration after transplantation in the degenerating retina. They also led to enhanced rescue of cell death and to preservation of electrophysiological function. Overall, we show that chemokines released from the degenerating retinae can drive migration of transplanted stem cells, and that overexpression of chemokine receptors can improve cell therapy-based regenerative approaches.


Asunto(s)
Células Madre Mesenquimatosas/metabolismo , Receptores CCR5/genética , Receptores CXCR6/genética , Degeneración Retiniana/etiología , Degeneración Retiniana/metabolismo , Animales , Biomarcadores , Movimiento Celular , Susceptibilidad a Enfermedades , Expresión Génica , Humanos , Ratones , Receptores CCR5/metabolismo , Receptores CXCR6/metabolismo , Degeneración Retiniana/patología
3.
Nucleic Acids Res ; 46(5): e30, 2018 03 16.
Artículo en Inglés | MEDLINE | ID: mdl-29294098

RESUMEN

CRISPR/dCas9-based labeling has allowed direct visualization of genomic regions in living cells. However, poor labeling efficiency and signal-to-background ratio have limited its application to visualize genome organization using super-resolution microscopy. We developed (Po)STAC (Polycistronic SunTAg modified CRISPR) by combining CRISPR/dCas9 with SunTag labeling and polycistronic vectors. (Po)STAC enhances both labeling efficiency and fluorescence signal detected from labeled loci enabling live cell imaging as well as super-resolution fixed-cell imaging of multiple genes with high spatiotemporal resolution.


Asunto(s)
Sistemas CRISPR-Cas/genética , Genes/genética , Vectores Genéticos/genética , Mediciones Luminiscentes/métodos , Imagen de Lapso de Tiempo/métodos , Animales , Línea Celular , Células Cultivadas , Células HEK293 , Células HeLa , Humanos , Hibridación Fluorescente in Situ/métodos , Proteínas Luminiscentes/genética , Proteínas Luminiscentes/metabolismo , Ratones , Reproducibilidad de los Resultados , Telómero/genética , Telómero/metabolismo
4.
Stem Cell Reports ; 19(5): 689-709, 2024 May 14.
Artículo en Inglés | MEDLINE | ID: mdl-38701778

RESUMEN

Embryo size, specification, and homeostasis are regulated by a complex gene regulatory and signaling network. Here we used gene expression signatures of Wnt-activated mouse embryonic stem cell (mESC) clones to reverse engineer an mESC regulatory network. We identify NKX1-2 as a novel master regulator of preimplantation embryo development. We find that Nkx1-2 inhibition reduces nascent RNA synthesis, downregulates genes controlling ribosome biogenesis, RNA translation, and transport, and induces severe alteration of nucleolus structure, resulting in the exclusion of RNA polymerase I from nucleoli. In turn, NKX1-2 loss of function leads to chromosome missegregation in the 2- to 4-cell embryo stages, severe decrease in blastomere numbers, alterations of tight junctions (TJs), and impairment of microlumen coarsening. Overall, these changes impair the blastocoel expansion-collapse cycle and embryo cavitation, leading to altered lineage specification and developmental arrest.


Asunto(s)
Desarrollo Embrionario , Regulación del Desarrollo de la Expresión Génica , Proteínas de Homeodominio , Factores de Transcripción , Animales , Ratones , Blastocisto/metabolismo , Blastocisto/citología , Nucléolo Celular/metabolismo , Desarrollo Embrionario/genética , Proteínas de Homeodominio/metabolismo , Proteínas de Homeodominio/genética , Células Madre Embrionarias de Ratones/metabolismo , Células Madre Embrionarias de Ratones/citología , Uniones Estrechas/metabolismo , Factores de Transcripción/metabolismo , Factores de Transcripción/genética , Proteínas Wnt/metabolismo , Vía de Señalización Wnt
5.
EBioMedicine ; 77: 103914, 2022 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-35278743

RESUMEN

BACKGROUND: Visual impairments are a critical medical hurdle to be addressed in modern society. Müller glia (MG) have regenerative potential in the retina in lower vertebrates, but not in mammals. However, in mice, in vivo cell fusion between MG and adult stem cells forms hybrids that can partially regenerate ablated neurons. METHODS: We used organotypic cultures of human retina and preparations of dissociated cells to test the hypothesis that cell fusion between human MG and adult stem cells can induce neuronal regeneration in human systems. Moreover, we established a microinjection system for transplanting human retinal organoids to demonstrate hybrid differentiation. FINDINGS: We first found that cell fusion occurs between MG and adult stem cells, in organotypic cultures of human retina as well as in cell cultures. Next, we showed that the resulting hybrids can differentiate and acquire a proto-neural electrophysiology profile when the Wnt/beta-catenin pathway is activated in the adult stem cells prior fusion. Finally, we demonstrated the engraftment and differentiation of these hybrids into human retinal organoids. INTERPRETATION: We show fusion between human MG and adult stem cells, and demonstrate that the resulting hybrid cells can differentiate towards neural fate in human model systems. Our results suggest that cell fusion-mediated therapy is a potential regenerative approach for treating human retinal dystrophies. FUNDING: This work was supported by La Caixa Health (HR17-00231), Velux Stiftung (976a) and the Ministerio de Ciencia e Innovación, (BFU2017-86760-P) (AEI/FEDER, UE), AGAUR (2017 SGR 689, 2017 SGR 926).


Asunto(s)
Células Madre Adultas , Células Ependimogliales , Animales , Diferenciación Celular , Células Ependimogliales/metabolismo , Humanos , Mamíferos , Ratones , Neuroglía , Retina/metabolismo
6.
FEBS J ; 286(6): 1074-1093, 2019 03.
Artículo en Inglés | MEDLINE | ID: mdl-30103260

RESUMEN

Regenerative capacities vary enormously across the animal kingdom. In contrast to most cold-blooded vertebrates, mammals, including humans, have very limited regenerative capacity when it comes to repairing damaged or degenerating tissues. Here, we review the main mechanisms of tissue regeneration, underlying the importance of cell dedifferentiation and reprogramming. We discuss the significance of cell fate and identity changes in the context of regenerative medicine, with a particular focus on strategies aiming at the promotion of the body's self-repairing mechanisms. We also introduce some of the most recent advances that have resulted in complete reprogramming of cell identity in vivo. Lastly, we discuss the main challenges that need to be addressed in the near future to develop in vivo reprogramming approaches with therapeutic potential.


Asunto(s)
Diferenciación Celular , Fusión Celular , Transdiferenciación Celular , Reprogramación Celular , Células Madre Pluripotentes Inducidas/citología , Medicina Regenerativa , Animales , Comunicación Celular , Humanos
7.
J Nutr Biochem ; 40: 187-193, 2017 02.
Artículo en Inglés | MEDLINE | ID: mdl-27915162

RESUMEN

After an acute myocardial infarction, obese patients generally have a better prognosis than their leaner counterparts, known as the "obesity paradox". In addition, female sex is associated with a lower risk of cardiac ischemic events and smaller infarct size compared to males. The objective of the present work was to study the metabolic phenotype and mitochondrial function associated to female sex and short-term high-fat diet. 1H NMR spectra of mice heart extracts were analysed by mRMR variable selection and linear discriminant analysis was used to evaluate metabolic changes. In separate experiments, O2 consumption and H2O2 production were measured from isolated mitochondria as well as serum oxidation susceptibility. Fingerprinting showed that male hearts contained more myo-inositol, taurine and glutamate than female hearts. HFD reduced the levels of creatine, taurine citrate and acetate. Profiling showed increased alanine and fumarate in HFD suggesting altered glycolitic and Krebs cycle pathways. Female mice contained less glucose than males. Female sex nor HFD altered mitochondria oxygen consumption but both conditions reduced the amount of H2O2 produced in an additive manner. Serum of females had lower oxidation susceptibility than serum from males but there were no differences associated with HFD. In conclusion, female sex and short-term HFD have an effect on the myocardial metabolic pattern and reduce the amount of H2O2 produced by mitochondria in an additive manner suggesting different mechanisms of action. This could explain, at least in part, the protection afforded by female sex and the "obesity paradox".


Asunto(s)
Dieta Alta en Grasa/efectos adversos , Miocardio/metabolismo , Estrés Oxidativo , Animales , Peso Corporal , Femenino , Peróxido de Hidrógeno/metabolismo , Lípidos/sangre , Espectroscopía de Resonancia Magnética , Masculino , Metabolómica/métodos , Ratones Endogámicos C57BL , Mitocondrias Cardíacas/metabolismo , Miocardio/patología , Consumo de Oxígeno , Factores Sexuales
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA