RESUMEN
Precipitation of nanocrystallites of cobalt-aluminium layered double hydroxides in a magnetic field has been studied. In a magnetic field perpendicular to the substrate, dense and homogeneous films have been obtained. Magnetic anisotropy of the crystallites is explained by deviation from the statistical cation distribution in favour of honeycomb-like coordination of cobalt.
RESUMEN
Multiferroic materials demonstrating coexistence of magnetic and ferroelectric orders are promising candidates for magnetoelectric devices. While understanding the underlying mechanism of interplaying of ferroic properties is important, tailoring their properties to make them potential candidates for magnetoelectric devices is challenging. Here, the antiferromagnetic Neel ordering temperature above 200 K is realized in successfully stabilized epitaxial films of (Lu,Sc)FeO3 multiferroic oxide. The first-principles calculations show the shrinkage of in-plane lattice constants of the unit cells of the films on different substrates which corroborates well the enhancement of the Neel ordering temperature (TN ). The profound effect of lattice strain/stress at the interface due to differences of in-plane lattice constants on out of plane magnetic properties and on spin reorientation temperature in the antiferromagnetic region is further elucidated in the epitaxial films with and without buffer layer of Mn-doped LuFeO3 . Writing and reading ferroelectric domains reveal the ferroelectric response of the films at room temperature. Detailed electron microscopy shows the presence of lattice defects in atomic scale. First-principles calculations show that orbital rehybridization of rare-earth ions and oxygen is one of the main driving force of ferroelectricity along c-axis in thin films of hexagonal ferrites.
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The lymphatic vasculature has an essential role in maintaining normal fluid balance in tissues and modulating the inflammatory response to injury or pathogens. Disruption of normal development or function of lymphatic vessels can have severe consequences. In the heart, reduced lymphatic function can lead to myocardial oedema and persistent inflammation. Macrophages, which are phagocytic cells of the innate immune system, contribute to cardiac development and to fibrotic repair and regeneration of cardiac tissue after myocardial infarction. In this Review, we discuss the cardiac lymphatic vasculature with a focus on developments over the past 5 years arising from the study of mammalian and zebrafish model organisms. In addition, we examine the interplay between the cardiac lymphatics and macrophages during fibrotic repair and regeneration after myocardial infarction. Finally, we discuss the therapeutic potential of targeting the cardiac lymphatic network to regulate immune cell content and alleviate inflammation in patients with ischaemic heart disease.
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Corazón , Inflamación , Vasos Linfáticos , Macrófagos , Isquemia Miocárdica , Regeneración , Animales , Modelos Animales de Enfermedad , Fibrosis/inmunología , Fibrosis/fisiopatología , Corazón/embriología , Corazón/fisiología , Corazón/fisiopatología , Humanos , Inflamación/inmunología , Inflamación/fisiopatología , Vasos Linfáticos/embriología , Vasos Linfáticos/inmunología , Vasos Linfáticos/fisiología , Vasos Linfáticos/fisiopatología , Macrófagos/inmunología , Macrófagos/fisiología , Isquemia Miocárdica/inmunología , Isquemia Miocárdica/fisiopatología , Miocardio/inmunología , Regeneración/inmunología , Regeneración/fisiologíaRESUMEN
The temperature behavior of the crystal structure as well as dielectric and magnetic properties of the perovskite bismuth chromate ceramics with the 10 mol % Cr3+-to-Sc3+ substitution were studied and compared with those of the unmodified compound. Using a high-pressure synthesis, BiCrO3 and BiCr0.9Sc0.1O3 were obtained as metastable perovskite phases which are monoclinic C2/c with the â6ap × â2ap × â6ap superstructure (where ap is the primitive perovskite unit-cell parameter) under ambient conditions. At room temperature, the unit cell volume of BiCr0.9Sc0.1O3 is â¼1.3% larger than that of BiCrO3. Both perovskites undergo a reversible structural transition into a nonpolar GdFeO3-type phase (orthorhombic Pnma, â2ap × 2ap × â2ap) in the temperature ranges of 410-420 K (BiCrO3) and 470-520 K (BiCr0.9Sc0.1O3) with a relative jump of the primitive perovskite unit cell volume of about -1.6 and -2.0%, respectively. Temperature dependences of the complex dielectric permittivity demonstrate anomalies in the phase transition ranges. The Pnma-to-C2/c crossover in BiCrO3 is accompanied by a decrease in the direct current (dc) conductivity, while in BiCr0.9Sc0.1O3 the conductivity increases. The onset of an antiferromagnetic order in BiCr0.9Sc0.1O3 is observed at the Néel temperature (TN) of about 85 K as compared with TN = 110 K in BiCrO3. In contrast to BiCrO3, which exhibits a spin reorientation at Tsr â¼ 72 K, no such a transition occurs in BiCr0.9Sc0.1O3.
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Canonical roles for macrophages in mediating the fibrotic response after a heart attack include extracellular matrix turnover and activation of cardiac fibroblasts to initiate collagen deposition. Here we reveal that macrophages directly contribute collagen to the forming post-injury scar. Unbiased transcriptomics shows an upregulation of collagens in both zebrafish and mouse macrophages following heart injury. Adoptive transfer of macrophages, from either collagen-tagged zebrafish or adult mouse GFPtpz-collagen donors, enhances scar formation via cell autonomous production of collagen. In zebrafish, the majority of tagged collagen localises proximal to the injury, within the overlying epicardial region, suggesting a possible distinction between macrophage-deposited collagen and that predominantly laid-down by myofibroblasts. Macrophage-specific targeting of col4a3bpa and cognate col4a1 in zebrafish significantly reduces scarring in cryoinjured hosts. Our findings contrast with the current model of scarring, whereby collagen deposition is exclusively attributed to myofibroblasts, and implicate macrophages as direct contributors to fibrosis during heart repair.
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Cicatriz/metabolismo , Cicatriz/patología , Colágeno/metabolismo , Corazón/fisiopatología , Macrófagos/patología , Cicatrización de Heridas , Pez Cebra/fisiología , Traslado Adoptivo , Animales , Embrión de Mamíferos/metabolismo , Proteínas de la Matriz Extracelular/metabolismo , Regulación de la Expresión Génica , Proteínas Fluorescentes Verdes/metabolismo , Macrófagos/metabolismo , Ratones , Monocitos/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Bazo/patología , Transcripción Genética , Transcriptoma/genética , Pez Cebra/embriología , Proteínas de Pez Cebra/metabolismoRESUMEN
Heart failure is a major cause of death worldwide owing to the inability of the adult human heart to regenerate after a heart attack. However, many vertebrate species are capable of complete cardiac regeneration following injury. In this Review, we discuss the various model organisms of cardiac regeneration, and outline what they have taught us thus far about the cellular and molecular responses essential for optimal cardiac repair. We compare across different species, highlighting evolutionarily conserved mechanisms of regeneration and demonstrating the importance of developmental gene expression programmes, plasticity of the heart and the pathophysiological environment for the regenerative response. Additionally, we discuss how the findings from these studies have led to improvements in cardiac repair in preclinical models such as adult mice and pigs, and discuss the potential to translate these findings into therapeutic approaches for human patients following myocardial infarction.
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Corazón/fisiología , Modelos Animales , Regeneración/fisiología , Animales , Humanos , Mamíferos , Modelos Biológicos , Medicina RegenerativaRESUMEN
In the adult rodent brain, new neurons are born in two germinal regions that are associated with blood vessels, and blood vessels and vessel-derived factors are thought to regulate the activity of adult neural stem cells. Recently, it has been proposed that a vascular niche also regulates prenatal neurogenesis. Here we identify the mouse embryo hindbrain as a powerful model to study embryonic neurogenesis and define the relationship between neural progenitor cell (NPC) behavior and vessel growth. Using this model, we show that a subventricular vascular plexus (SVP) extends through a hindbrain germinal zone populated by NPCs whose peak mitotic activity follows a surge in SVP growth. Hindbrains genetically defective in SVP formation owing to constitutive NRP1 loss showed a premature decline in both NPC activity and hindbrain growth downstream of precocious cell cycle exit, premature neuronal differentiation, and abnormal mitosis patterns. Defective regulation of NPC activity was not observed in mice lacking NRP1 expression by NPCs, but instead in mice lacking NRP1 selectively in endothelial cells, yet was independent of vascular roles in hindbrain oxygenation. Therefore, germinal zone vascularization sustains NPC proliferation in the prenatal brain.
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Vasos Sanguíneos/fisiología , Neurogénesis , Rombencéfalo/irrigación sanguínea , Rombencéfalo/embriología , Animales , Proliferación Celular , Autorrenovación de las Células , Células Endoteliales/metabolismo , Matriz Extracelular/metabolismo , Ratones , Ratones Endogámicos C57BL , Mitosis , Neovascularización Fisiológica , Células-Madre Neurales/citología , Células-Madre Neurales/metabolismo , Neuropilina-1/metabolismo , Oxígeno/metabolismo , Factores de TiempoRESUMEN
Correct regulation of troponin and myosin contractile protein gene isoforms is a critical determinant of cardiac and skeletal striated muscle development and function, with misexpression frequently associated with impaired contractility or disease. Here we reveal a novel requirement for Prospero-related homeobox factor 1 (Prox1) during mouse heart development in the direct transcriptional repression of the fast-twitch skeletal muscle genes troponin T3, troponin I2, and myosin light chain 1. A proportion of cardiac-specific Prox1 knockout mice survive beyond birth with hearts characterized by marked overexpression of fast-twitch genes and postnatal development of a fatal dilated cardiomyopathy. Through conditional knockout of Prox1 from skeletal muscle, we demonstrate a conserved requirement for Prox1 in the repression of troponin T3, troponin I2, and myosin light chain 1 between cardiac and slow-twitch skeletal muscle and establish Prox1 ablation as sufficient to cause a switch from a slow- to fast-twitch muscle phenotype. Our study identifies conserved roles for Prox1 between cardiac and skeletal muscle, specifically implicated in slow-twitch fiber-type specification, function, and cardiomyopathic disease.
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Cardiomiopatía Dilatada/metabolismo , Regulación del Desarrollo de la Expresión Génica/fisiología , Músculo Estriado/embriología , Miocardio/metabolismo , Proteínas Supresoras de Tumor/deficiencia , Animales , Diferenciación Celular/fisiología , Inmunoprecipitación de Cromatina , Cartilla de ADN/genética , Técnica del Anticuerpo Fluorescente , Proteínas de Homeodominio/genética , Ratones , Ratones Noqueados , Análisis por Micromatrices , Músculo Estriado/metabolismo , Cadenas Ligeras de Miosina/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Troponina/metabolismo , Troponina I/metabolismo , Proteínas Supresoras de Tumor/genéticaRESUMEN
The mouse embryo hindbrain is a robust and adaptable model for studying sprouting angiogenesis. It permits the spatiotemporal analysis of organ vascularization in normal mice and in mouse strains with genetic mutations that result in late embryonic or perinatal lethality. Unlike postnatal models such as retinal angiogenesis or Matrigel implants, there is no requirement for the breeding of conditional knockout mice. The unique architecture of the hindbrain vasculature allows whole-mount immunolabeling of blood vessels and high-resolution imaging, as well as easy quantification of angiogenic sprouting, network density and vessel caliber. The hindbrain model also permits the visualization of ligand binding to blood vessels in situ and the analysis of blood vessel growth within a natural multicellular microenvironment in which endothelial cells (ECs) interact with non-ECs to refine the 3D organ architecture. The entire procedure, from embryo isolation to imaging and through to results analysis, takes approximately 4 d.
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Diagnóstico por Imagen/métodos , Embrión de Mamíferos/embriología , Modelos Animales , Neovascularización Fisiológica/fisiología , Rombencéfalo/embriología , Animales , Anticuerpos Monoclonales , Ligandos , Ratones , Rombencéfalo/irrigación sanguíneaRESUMEN
Neuropilin (NRP) 1 is a receptor for the vascular endothelial growth factor (VEGF)-A and is essential for normal angiogenesis. Previous in vitro experiments identified NRP1 interactions with VEGF-A's main signaling receptor VEGFR2 within endothelial cells, but also between nonendothelial NRP1 and endothelial VEGFR2. Consistent with an endothelial role for NRP1 in angiogenesis, we found that VEGFR2 and NRP1 were coexpressed in endothelial tip and stalk cells in the developing brain. In addition, NRP1 was expressed on two cell types that interact with growing brain vessels-the neural progenitors that secrete VEGF-A to stimulate tip cell activity and the pro-angiogenic macrophages that promote tip cell anastomosis. Selective targeting of Nrp1 in each of these cell types demonstrated that neural progenitor- and macrophage-derived NRP1 were dispensable, whereas endothelial NRP1 was essential for normal brain vessel growth. NRP1 therefore promotes brain angiogenesis cell autonomously in endothelium, independently of heterotypic interactions with nonendothelial cells. Genetic mosaic analyses demonstrated a key role for NRP1 in endothelial tip rather than stalk cells during vessel sprouting. Thus, NRP1-expressing endothelial cells attained the tip cell position when competing with NRP1-negative endothelial cells in chimeric vessel sprouts. Taken together, these findings demonstrate that NRP1 promotes endothelial tip cell function during angiogenesis.
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Vasos Sanguíneos/embriología , Células Endoteliales/fisiología , Endotelio Vascular/citología , Neovascularización Fisiológica/genética , Neuropilina-1/fisiología , Animales , Vasos Sanguíneos/crecimiento & desarrollo , Vasos Sanguíneos/metabolismo , Polaridad Celular/genética , Embrión de Mamíferos , Células Endoteliales/citología , Células Endoteliales/metabolismo , Endotelio Vascular/metabolismo , Femenino , Ratones , Ratones Transgénicos , Modelos Biológicos , Morfogénesis/genética , Neuropilina-1/genética , Neuropilina-1/metabolismo , Especificidad de Órganos/genética , EmbarazoRESUMEN
Over the last decade or so, intensive research in cardiac stem cell biology has led to significant discoveries towards a potential therapy for cardiovascular disease; the main cause of morbidity and mortality in humans. The major goal within the field of cardiovascular regenerative medicine is to replace lost or damaged cardiac muscle and coronaries following ischaemic disease. At present, de novo cardiomyocytes can be generated either in vitro, for cell transplantation or disease modelling using directed differentiation of embryonic stem cells or induced pluripotent stem cells, or in vivo via direct reprogramming of resident adult cardiac fibroblast or ectopic stimulation of resident cardiac stem or progenitor cells. A major bottleneck with all of these approaches is the low efficiency of cardiomyocyte differentiation alongside their relative functional immaturity. Chemical genetics, and the application of phenotypic screening with small molecule libraries, represent a means to enhance understanding of the molecular pathways controlling cardiovascular cell differentiation and, moreover, offer the potential for discovery of new drugs to invoke heart repair and regeneration. Here, we review the potential of chemical genetics in cardiac stem cell therapy, highlighting not only the major contributions to the field so far, but also the future challenges.
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Enfermedades Cardiovasculares/terapia , Miocitos Cardíacos/citología , Trasplante de Células Madre/métodos , Adulto , Animales , Enfermedades Cardiovasculares/fisiopatología , Diferenciación Celular/fisiología , Descubrimiento de Drogas/métodos , Células Madre Embrionarias/citología , Fibroblastos/citología , Humanos , Células Madre Pluripotentes Inducidas/citología , Regeneración/fisiología , Medicina Regenerativa/métodos , Medicina Regenerativa/tendencias , Bibliotecas de Moléculas Pequeñas/farmacología , Trasplante de Células Madre/tendenciasRESUMEN
Efficient cardiac regeneration postinfarction (MI) requires the replacement of lost cardiomyocytes, formation of new coronary vessels and appropriate modulation of the inflammatory response. However, insight into how to stimulate repair of the human heart is currently limited. Using the embryonic paradigm of regeneration, we demonstrated that the actin-binding peptide thymosin ß4 (Tß4), required for epicardium-derived coronary vasculogenesis, can recapitulate its embryonic role and activate quiescent adult epicardial cells (EPDCs). Once stimulated, EPDCs facilitate neovascularization of the ischemic adult heart and, moreover, contribute bona fide cardiomyocytes. EPDC-derived cardiomyocytes structurally and functionally integrate with resident muscle to regenerate functional myocardium, limiting pathological remodeling, and effecting an improvement in cardiac function. Alongside pro-survival and anti-inflammatory properties, these regenerative roles, via EPDCs, markedly expand the range of therapeutic benefits of Tß4 to sustain and repair the myocardium after ischemic damage.
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Isquemia Miocárdica/metabolismo , Miocardio/metabolismo , Timosina/metabolismo , Animales , Humanos , Miocardio/patología , Miocitos Cardíacos/metabolismo , Pericardio/metabolismo , Pericardio/patología , Regeneración/fisiologíaRESUMEN
RATIONALE: Nkx2.5 is one of the most widely studied cardiac-specific transcription factors, conserved from flies to man, with multiple essential roles in both the developing and adult heart. Specific dominant mutations in NKX2.5 have been identified in adult congenital heart disease patients presenting with conduction system anomalies and recent genome-wide association studies implicate the NKX2.5 locus, as causative for lethal arrhythmias ("sudden cardiac death") that occur at a frequency in the population of 1 in 1000 per annum worldwide. Haploinsufficiency for Nkx2.5 in the mouse phenocopies human conduction disease pathology yet the phenotypes, described in both mouse and man, are highly pleiotropic, implicit of unknown modifiers and/or factors acting in epistasis with Nkx2.5/NKX2.5. OBJECTIVE: To identify bone fide upstream genetic modifier(s) of Nkx2.5/NKX2.5 function and to determine epistatic effects relevant to the manifestation of NKX2.5-dependent adult congenital heart disease. METHODS AND RESULTS: A study of cardiac function in prospero-related homeobox protein 1 (Prox1) heterozygous mice, using pressure-volume loop and micromannometry, revealed rescue of hemodynamic parameters in Nkx2.5(Cre/+); Prox1(loxP/+) animals versus Nkx2.5(Cre/+) controls. Anatomic studies, on a Cx40(EGFP) background, revealed Cre-mediated knock-down of Prox1 restored the anatomy of the atrioventricular node and His-Purkinje network both of which were severely hypoplastic in Nkx2.5(Cre/+) littermates. Steady state surface electrocardiography recordings and high-speed multiphoton imaging, to assess Ca(2+) handling, revealed atrioventricular conduction and excitation-contraction were also normalized by Prox1 haploinsufficiency, as was expression of conduction genes thought to act downstream of Nkx2.5. Chromatin immunoprecipitation on adult hearts, in combination with both gain and loss-of-function reporter assays in vitro, revealed that Prox1 recruits the corepressor HDAC3 to directly repress Nkx2.5 via a proximal upstream enhancer as a mechanism for regulating Nkx2.5 function in adult cardiac conduction. CONCLUSIONS: Here we identify Prox1 as a direct upstream modifier of Nkx2.5 in the maintenance of the adult conduction system and rescue of Nkx2.5 conduction disease phenotypes. This study is the first example of rescue of Nkx2.5 function and establishes a model for ensuring electrophysiological function within the adult heart alongside insight into a novel Prox1-HDAC3-Nkx2.5 signaling pathway for therapeutic targeting in conduction disease.
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Epistasis Genética/genética , Sistema de Conducción Cardíaco/fisiopatología , Cardiopatías/genética , Cardiopatías/metabolismo , Histona Desacetilasas/genética , Proteínas de Homeodominio/genética , Fenotipo , Factores de Transcripción/genética , Proteínas Supresoras de Tumor/genética , Animales , Cardiopatías/fisiopatología , Histona Desacetilasas/fisiología , Proteína Homeótica Nkx-2.5 , Proteínas de Homeodominio/fisiología , Ratones , Ratones Transgénicos , Células 3T3 NIH , Factores de Transcripción/fisiología , Proteínas Supresoras de Tumor/fisiologíaRESUMEN
A significant bottleneck in cardiovascular regenerative medicine is the identification of a viable source of stem/progenitor cells that could contribute new muscle after ischaemic heart disease and acute myocardial infarction. A therapeutic ideal--relative to cell transplantation--would be to stimulate a resident source, thus avoiding the caveats of limited graft survival, restricted homing to the site of injury and host immune rejection. Here we demonstrate in mice that the adult heart contains a resident stem or progenitor cell population, which has the potential to contribute bona fide terminally differentiated cardiomyocytes after myocardial infarction. We reveal a novel genetic label of the activated adult progenitors via re-expression of a key embryonic epicardial gene, Wilm's tumour 1 (Wt1), through priming by thymosin ß4, a peptide previously shown to restore vascular potential to adult epicardium-derived progenitor cells with injury. Cumulative evidence indicates an epicardial origin of the progenitor population, and embryonic reprogramming results in the mobilization of this population and concomitant differentiation to give rise to de novo cardiomyocytes. Cell transplantation confirmed a progenitor source and chromosome painting of labelled donor cells revealed transdifferentiation to a myocyte fate in the absence of cell fusion. Derived cardiomyocytes are shown here to structurally and functionally integrate with resident muscle; as such, stimulation of this adult progenitor pool represents a significant step towards resident-cell-based therapy in human ischaemic heart disease.
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Células Madre Adultas/citología , Diferenciación Celular , Lesiones Cardíacas , Miocitos Cardíacos/citología , Animales , Reprogramación Celular , Regulación de la Expresión Génica , Ratones , Infarto del Miocardio/patología , Miocitos Cardíacos/metabolismo , Timosina/metabolismo , Proteínas WT1/genética , Proteínas WT1/metabolismoRESUMEN
During development, the axons of retinal ganglion cell (RGC) neurons must decide whether to cross or avoid the midline at the optic chiasm to project to targets on both sides of the brain. By combining genetic analyses with in vitro assays, we show that neuropilin 1 (NRP1) promotes contralateral RGC projection in mammals. Unexpectedly, the NRP1 ligand involved is not an axon guidance cue of the class 3 semaphorin family, but VEGF164, the neuropilin-binding isoform of the classical vascular growth factor VEGF-A. VEGF164 is expressed at the chiasm midline and is required for normal contralateral growth in vivo. In outgrowth and growth cone turning assays, VEGF164 acts directly on NRP1-expressing contralateral RGCs to provide growth-promoting and chemoattractive signals. These findings have identified a permissive midline signal for axons at the chiasm midline and provide in vivo evidence that VEGF-A is an essential axon guidance cue.
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Regulación del Desarrollo de la Expresión Génica/fisiología , Neuropilina-1/metabolismo , Quiasma Óptico/fisiología , Transducción de Señal/fisiología , Factores de Crecimiento Endotelial Vascular/metabolismo , Factores de Edad , Aminoácidos/metabolismo , Animales , Axones/efectos de los fármacos , Embrión de Mamíferos , Lateralidad Funcional , Regulación del Desarrollo de la Expresión Génica/genética , Conos de Crecimiento/fisiología , Indoles/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Neuropilina-1/deficiencia , Neuropilina-1/genética , Neuropilina-2/deficiencia , Quiasma Óptico/efectos de los fármacos , Quiasma Óptico/embriología , Técnicas de Cultivo de Órganos , Retina/citología , Retina/efectos de los fármacos , Retina/embriología , Células Ganglionares de la Retina/citología , Células Ganglionares de la Retina/efectos de los fármacos , Células Ganglionares de la Retina/fisiología , Semaforina-3A/deficiencia , Transducción de Señal/genética , Factor A de Crecimiento Endotelial Vascular/genética , Factor A de Crecimiento Endotelial Vascular/farmacología , Receptor 1 de Factores de Crecimiento Endotelial Vascular/genética , Receptor 2 de Factores de Crecimiento Endotelial Vascular/genética , Factores de Crecimiento Endotelial Vascular/genéticaRESUMEN
Cardiovascular regenerative medicine aims to counter muscle loss post ischaemic disease with the identification of new cellular sources for cardiomyocyte replacement. A number of embryonic and adult cell models have been explored preclinically and in patient trials, but modest outcome, coupled with issues with impaired graft survival and limited/immature (trans-) differentiation alongside host rejection, has left the door open for more therapeutically efficacious sources of myocardial regeneration. Due to its fundamental role in heart development, the epicardium emerges as an obvious candidate. Here, recent findings are reviewed that show adult epicardium-derived cells as a new source of regenerative capacity for heart repair.
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Células Madre Adultas/trasplante , Cardiomiopatías/terapia , Pericardio/citología , Medicina Regenerativa/métodos , Trasplante de Células Madre/métodos , Adulto , Células Madre Embrionarias/trasplante , Humanos , Pericardio/trasplanteRESUMEN
Blood vessel networks expand in a 2-step process that begins with vessel sprouting and is followed by vessel anastomosis. Vessel sprouting is induced by chemotactic gradients of the vascular endothelial growth factor (VEGF), which stimulates tip cell protrusion. Yet it is not known which factors promote the fusion of neighboring tip cells to add new circuits to the existing vessel network. By combining the analysis of mouse mutants defective in macrophage development or VEGF signaling with live imaging in zebrafish, we now show that macrophages promote tip cell fusion downstream of VEGF-mediated tip cell induction. Macrophages therefore play a hitherto unidentified and unexpected role as vascular fusion cells. Moreover, we show that there are striking molecular similarities between the pro-angiogenic tissue macrophages essential for vascular development and those that promote the angiogenic switch in cancer, including the expression of the cell-surface proteins TIE2 and NRP1. Our findings suggest that tissue macrophages are a target for antiangiogenic therapies, but that they could equally well be exploited to stimulate tissue vascularization in ischemic disease.
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Macrófagos/fisiología , Neovascularización Fisiológica/efectos de los fármacos , Rombencéfalo/irrigación sanguínea , Factor A de Crecimiento Endotelial Vascular/fisiología , Animales , Polaridad Celular , Células Endoteliales/efectos de los fármacos , Células Endoteliales/fisiología , Células Endoteliales/ultraestructura , Endotelio Vascular/crecimiento & desarrollo , Femenino , Técnicas de Sustitución del Gen , Factor Estimulante de Colonias de Macrófagos/deficiencia , Factor Estimulante de Colonias de Macrófagos/genética , Factor Estimulante de Colonias de Macrófagos/fisiología , Masculino , Ratones , Ratones Noqueados , Neovascularización Fisiológica/fisiología , Neuropilina-1/fisiología , Proteínas Proto-Oncogénicas/deficiencia , Proteínas Tirosina Quinasas Receptoras/fisiología , Receptor TIE-2 , Vasos Retinianos/crecimiento & desarrollo , Rombencéfalo/embriología , Transactivadores/deficiencia , Factor A de Crecimiento Endotelial Vascular/deficiencia , Factor A de Crecimiento Endotelial Vascular/genética , Saco Vitelino/citología , Pez Cebra/embriologíaRESUMEN
Neural crest cells (NCCs) are highly motile embryonic stem cells that delaminate from the neuroectoderm early during vertebrate embryogenesis and differentiate at defined target sites into various essential cell types. To reach their targets, NCCs follow 1 of 3 sequential pathways that correlate with NCC fate. The firstborn NCCs travel ventrally alongside intersomitic blood vessels to form sympathetic neuronal progenitors near the dorsal aorta, while the lastborn NCCs migrate superficially beneath the epidermis to give rise to melanocytes. Yet, most NCCs enter the somites to form the intermediate wave that gives rise to sympathetic and sensory neurons. Here we show that the repulsive guidance cue SEMA3A and its receptor neuropilin 1 (NRP1) are essential to direct the intermediate wave NCC precursors of peripheral neurons from a default pathway alongside intersomitic blood vessels into the anterior sclerotome. Thus, loss of function for either gene caused excessive intersomitic NCC migration, and this led to ectopic neuronal differentiation along both the anteroposterior and dorsoventral axes of the trunk. The choice of migratory pathway did not affect the specification of NCCs, as they retained their commitment to differentiate into sympathetic or sensory neurons, even when they migrated on an ectopic dorsolateral path that is normally taken by melanocyte precursors. We conclude that NRP1 signaling coordinates pathway choice with NCC fate and therefore confines neuronal differentiation to appropriate locations.
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Movimiento Celular , Cresta Neural/metabolismo , Neuropilina-1/metabolismo , Transducción de Señal , Animales , Femenino , Regulación del Desarrollo de la Expresión Génica , Masculino , Ratones , Cresta Neural/irrigación sanguínea , Cresta Neural/citología , Cresta Neural/embriología , Neuropilina-1/genética , Semaforina-3A/genética , Semaforina-3A/metabolismoRESUMEN
Neuropilin (NRP) receptors and their class 3 semaphorin (SEMA3) ligands play well-established roles in axon guidance, with loss of NRP1, NRP2, SEMA3A or SEMA3F causing defasciculation and errors in growth cone guidance of peripherally projecting nerves. Here we report that loss of NRP1 or NRP2 also impairs sensory neuron positioning in the mouse head, and that this defect is a consequence of inappropriate cranial neural crest cell migration. Specifically, neural crest cells move into the normally crest-free territory between the trigeminal and hyoid neural crest streams and recruit sensory neurons from the otic placode; these ectopic neurons then extend axons between the trigeminal and facioacoustic ganglia. Moreover, we found that NRP1 and NRP2 cooperate to guide cranial neural crest cells and position sensory neurons; thus, in the absence of SEMA3/NRP signalling, the segmentation of the cranial nervous system is lost. We conclude that neuropilins play multiple roles in the sensory nervous system by directing cranial neural crest cells, positioning sensory neurons and organising their axonal projections.