RESUMO
The impressive morphological diversification of vertebrates was achieved in part by innovation and modification of the pharyngeal skeleton. Extensive fate mapping in amniote models has revealed a primarily cranial neural crest derivation of the pharyngeal skeleton. Although comparable fate maps of amphibians produced over several decades have failed to document a neural crest derivation of ventromedial elements in these vertebrates, a recent report provides evidence of a mesodermal origin of one of these elements, basibranchial 2, in the axolotl. We used a transgenic labeling protocol and grafts of labeled cells between GFP+ and white embryos to derive a fate map that describes contributions of both cranial neural crest and mesoderm to the axolotl pharyngeal skeleton, and we conducted additional experiments that probe the mechanisms that underlie mesodermal patterning. Our fate map confirms a dual embryonic origin of the pharyngeal skeleton in urodeles, including derivation of basibranchial 2 from mesoderm closely associated with the second heart field. Additionally, heterotopic transplantation experiments reveal lineage restriction of mesodermal cells that contribute to pharyngeal cartilage. The mesoderm-derived component of the pharyngeal skeleton appears to be particularly sensitive to retinoic acid (RA): administration of exogenous RA leads to loss of the second basibranchial, but not the first. Neural crest was undoubtedly critical in the evolution of the vertebrate pharyngeal skeleton, but mesoderm may have played a central role in forming ventromedial elements, in particular. When and how many times during vertebrate phylogeny a mesodermal contribution to the pharyngeal skeleton evolved remain to be resolved.
Assuntos
Ambystoma mexicanum/embriologia , Evolução Biológica , Padronização Corporal , Osso e Ossos/embriologia , Faringe/embriologia , Ambystoma mexicanum/genética , Animais , Embrião não Mamífero/metabolismo , Mesoderma/embriologia , Crista Neural/embriologia , Tretinoína/metabolismoRESUMO
The Zmiz2 (Zimp7) protein and its homolog Zmiz1 (Zimp10) were initially identified in humans as androgen receptor co-activators. Sequence analysis revealed the presence of an SP-RING/Miz domain, which is highly conserved in members of the PIAS family and confers SUMO-conjugating activity. Zimp7 has been shown to interact with components of the Wnt/ß-Catenin signaling pathway and with Brg1 and BAF57, components of the ATP-dependent mammalian SWI/SNF-like BAF chromatin-remodeling complexes. In this work, we analyze the role of zygotic Zimp7 in zebrafish development. We describe evidence indicating that Zimp7 is required for mesoderm development and dorsoventral patterning. Morpholino-mediated reduction of zygotic Zimp7 produced axial mesodermal defects that were preceded by up-regulation of organizer genes such as bozozok, goosecoid and floating head at the onset of gastrulation and by down-regulation of the ventral markers vox, vent and eve1 indicating loss of the ventrolateral mesoderm. Consistently, embryos overexpressing zimp7 RNA exhibited midline defects such as loss of forebrain and cyclopia accompanied by transcriptional changes directly opposite of those found in the morphants. In addition, the patterning of ventralized embryos produced by the overexpression of vox and vent was restored by a reduction of Zimp7 activity. Altogether, our findings indicate that Zimp7 is involved in transcriptional regulation of factors that are essential for patterning in the dorsoventral axis.
Assuntos
Padronização Corporal/genética , Regulação da Expressão Gênica no Desenvolvimento , Organizadores Embrionários/embriologia , Proteínas Inibidoras de STAT Ativados/metabolismo , Proteínas de Peixe-Zebra/metabolismo , Peixe-Zebra/embriologia , Dedos de Zinco/genética , Animais , Blástula/metabolismo , Gastrulação/genética , Técnicas de Silenciamento de Genes , Proteína Goosecoid/biossíntese , Proteínas de Homeodomínio/biossíntese , Mesoderma/embriologia , Morfolinos/genética , Proteínas Inibidoras de STAT Ativados/genética , RNA Mensageiro/biossíntese , Proteínas Repressoras/biossíntese , Transativadores/genética , Fatores de Transcrição/biossíntese , Transcrição Gênica/genética , Proteínas de Peixe-Zebra/biossíntese , Proteínas de Peixe-Zebra/genéticaRESUMO
In vertebrates, the embryonic dorsal midline is a crucial signalling centre that patterns the surrounding tissues during development. Members of the FoxA subfamily of transcription factors are expressed in the structures that compose this centre. Foxa2 is essential for dorsal midline development in mammals, since knock-out mouse embryos lack a definitive node, notochord and floor plate. The related gene foxA4 is only present in amphibians. Expression begins in the blastula -chordin and -noggin expressing centre (BCNE) and is later restricted to the dorsal midline derivatives of the Spemann's organiser. It was suggested that the early functions of mammalian foxa2 are carried out by foxA4 in frogs, but functional experiments were needed to test this hypothesis. Here, we show that some important dorsal midline functions of mammalian foxa2 are exerted by foxA4 in Xenopus. We provide new evidence that the latter prevents the respecification of dorsal midline precursors towards contiguous fates, inhibiting prechordal and paraxial mesoderm development in favour of the notochord. In addition, we show that foxA4 is required for the correct regionalisation and maintenance of the central nervous system. FoxA4 participates in constraining the prospective rostral forebrain territory during neural specification and is necessary for the correct segregation of the most anterior ectodermal derivatives, such as the cement gland and the pituitary anlagen. Moreover, the early expression of foxA4 in the BCNE (which contains precursors of the whole forebrain and most of the midbrain and hindbrain) is directly required to restrict anterior neural development.
Assuntos
Sistema Nervoso Central/embriologia , Sistema Nervoso Central/metabolismo , Embrião não Mamífero/metabolismo , Fatores de Transcrição Forkhead/metabolismo , Mesoderma/embriologia , Notocorda/embriologia , Proteínas de Xenopus/metabolismo , Xenopus/embriologia , Animais , Apoptose/efeitos dos fármacos , Biomarcadores/metabolismo , Blástula/efeitos dos fármacos , Blástula/metabolismo , Padronização Corporal/efeitos dos fármacos , Embrião não Mamífero/efeitos dos fármacos , Desenvolvimento Embrionário/efeitos dos fármacos , Técnicas de Silenciamento de Genes , Glicoproteínas/metabolismo , Cabeça/anormalidades , Cabeça/embriologia , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Mesoderma/efeitos dos fármacos , Mesoderma/metabolismo , Modelos Biológicos , Morfogênese/efeitos dos fármacos , Morfolinos/farmacologia , Placa Neural/embriologia , Placa Neural/metabolismo , Neurogênese/efeitos dos fármacos , Notocorda/efeitos dos fármacos , Notocorda/metabolismo , Fenótipo , Xenopus/metabolismoRESUMO
Maternal malnutrition due to a low-protein diet is associated with functional disorders in adulthood, which may be related to embryonic development failures. The effects of gestational protein restriction on prostate morphogenesis in male offspring were investigated. Pregnant rat dams were divided into normoprotein (NP; fed a normal diet containing 17% protein) and hypoprotein (LP; fed a diet containing 6% protein) groups. On the day of birth (PND1), anogenital distance and bodyweight were measured in male pups. Seven males per experimental group (one male per litter) were killed, and the pelvic urethra was evaluated. LP offspring showed a significant reduction in bodyweight and anogenital distance on PND1. On three-dimensional reconstruction of the prostate, the number of prostatic buds was lower in LP than in NP males. Mesenchymal cells surrounding the buds were androgen-receptor positive, and the quantity and intensity of nucleus immunoreactivity was decreased in LP. The proliferation index was lower in LP than in NP prostatic buds. Immunoreactivity for α-actin in mesenchymal cells and that for epidermal growth factor receptor in epithelial cells was higher in NP than in LP. Our findings demonstrate that maternal protein restriction delays prostatic morphogenesis, probably because of considerable disruption in the epithelium-mesenchyme interaction.
Assuntos
Organogênese/fisiologia , Complicações na Gravidez , Próstata/embriologia , Deficiência de Proteína/complicações , Animais , Animais Recém-Nascidos , Apoptose , Proliferação de Células , Dieta com Restrição de Proteínas , Células Epiteliais/citologia , Feminino , Masculino , Mesoderma/química , Mesoderma/embriologia , Gravidez , Complicações na Gravidez/etiologia , Próstata/citologia , Ratos , Ratos Wistar , Receptores Androgênicos/análiseRESUMO
Sipuncula is an ancient clade of unsegmented marine worms that develop through a conserved pattern of unequal quartet spiral cleavage. They exhibit putative character modifications, including conspicuously large first-quartet micromeres and prototroch cells, postoral metatroch with exclusive locomotory function, paired retractor muscles and terminal organ system, and a U-shaped digestive architecture with left-right asymmetric development. Four developmental life history patterns are recognized, and they have evolved a unique metazoan larval type, the pelagosphera. When compared with other quartet spiral-cleaving models, sipunculan development is understudied, challenging and typically absent from evolutionary interpretations of spiralian larval and adult body plan diversity. If spiral cleavage is appropriately viewed as a flexible character complex, then understudied clades and characters should be investigated. We are pursuing sipunculan models for modern molecular, genetic and cellular research on evolution of spiralian development. Protocols for whole mount gene expression studies are established in four species. Molecular labeling and confocal imaging techniques are operative from embryogenesis through larval development. Next-generation sequencing of developmental transcriptomes has been completed for two species with highly contrasting life history patterns, Phascolion cryptum (direct development) and Nephasoma pellucidum (indirect planktotrophy). Looking forward, we will attempt intracellular lineage tracing and fate-mapping studies in a proposed model sipunculan, Themiste lageniformis. Importantly, with the unsegmented Sipuncula now repositioned within the segmented Annelida, sipunculan worms have become timely and appropriate models for investigating the potential for flexibility in spiralian development, including segmentation. We briefly review previous studies, and discuss new observations on the spiralian character complex within Sipuncula.
Assuntos
Padronização Corporal/fisiologia , Embrião não Mamífero/embriologia , Larva/crescimento & desenvolvimento , Poliquetos/embriologia , Transcriptoma/genética , Animais , Evolução Biológica , Linhagem da Célula , Ectoderma/embriologia , Endoderma/embriologia , Larva/citologia , Mesoderma/embriologia , Partenogênese/fisiologia , Poliquetos/anatomia & histologia , Poliquetos/crescimento & desenvolvimentoRESUMO
Traditionally, the cartilaginous viscerocranium of vertebrates is considered as neural crest (NC)-derived. Morphological work carried out on amphibian embryos in the first half of the XX century suggested potentially mesodermal origin for some hyobranchial elements. Since then, the embryonic sources of the hyobranchial apparatus in amphibians has not been investigated due to lack of an appropriate long-term labelling system. We performed homotopic transplantations of neural folds along with the majority of cells of the presumptive NC, and/or fragments of the head lateral plate mesoderm (LPM) from transgenic GFP+ into white embryos. In these experiments, the NC-derived GFP+ cells contributed to all hyobranchial elements, except for basibranchial 2, whereas the grafting of GFP+ head mesoderm led to a reverse labelling result. The grafting of only the most ventral part of the head LPM resulted in marking of the basibranchial 2 and the heart myocardium, implying their origin from a common mesodermal region. This is the first evidence of contribution of LPM of the head to cranial elements in any vertebrate. If compared to fish, birds, and mammals, in which all branchial skeletal elements are NC-derived, the axolotl (probably this is true for all amphibians) demonstrates an evolutionary deviation, in which the head LPM replaces NC cells in a hyobranchial element. This implies that cells of different embryonic origin may have the same developmental program, leading to the formation of identical (homologous) elements of the skeleton.
Assuntos
Ambystoma mexicanum/embriologia , Ambystoma mexicanum/fisiologia , Animais , Animais Geneticamente Modificados , Aves , Osso e Ossos/embriologia , Cartilagem/embriologia , Peixes , Proteínas de Fluorescência Verde/química , Proteínas de Fluorescência Verde/metabolismo , Cabeça/embriologia , Coração/embriologia , Mesoderma/embriologia , Mesoderma/metabolismo , Mesoderma/fisiologia , Miocárdio/patologia , Crista Neural/metabolismo , Crista Neural/patologiaRESUMO
El aparato digestivo deriva del endodermo y el mesodermo, que forman su epitelio y la musculatura lisa respectivamente. Al igual que en el resto de los sistemas, existe un interacción epitelio-mesenquimática mediada por moléculas como Hedgehog, BMP y FoxF1 que determinan el crecimiento intestinal en sus ejes principales. Los genes Hox, junto con el resto de las moléculas, participan en la regionalización del sistema digestivo. En sus inicios lo denominaremos intestino primitivo, formado por un tubo endodérmico que deriva del saco vitelino; dividiéndose en intestino anterior, medio y posterior. En esta revisión veremos cómo estos 3 segmentos darán origen a las diferentes estructuras del sistema digestivo en los vertebrados.
The digestive system is derived from the endoderm and mesoderm, which form its epithelium and smooth muscle, respectively. As in the other systems, there is an epithelial-mesenchymal interactions mediated by molecules such as Hedgehog, BMP and FoxF1, determining intestinal growth in the main axes. The Hox genes, together the rest of the molecules, involved in the regionalization of the digestive system. In the beginning we call it primitive gut, consisting of a tube derived of endodermal yolk sac, divided into foregut, midgut and hindgut. In this review we will see how these 3 segments give rise to different structures of the digestive system in vertebrates.
Assuntos
Humanos , Animais , Sistema Digestório/embriologia , Vertebrados , Genes Homeobox , Proteínas Morfogenéticas Ósseas , Sistema Digestório/crescimento & desenvolvimento , Endoderma/embriologia , Proteínas Hedgehog , Mesoderma/embriologiaRESUMO
As células tronco mesenquimais derivadas do âmnio (AMSCs) são células multipotentes com alto potencial para se diferenciar em múltiplas linhagens. Podem ser isoladas sem recurso a procedimentos invasivos e usadas sem levantar quaisquer implicações éticas. O presente estudo visa isolar e caracterizar as células mesenquimais progenitoras da membrana amniótica de gatos domésticos para futura aplicação em terapia celular. As células foram isoladas de quatro membranas fetais, coletadas durante as campanhas rotineiras de castração em gatas no último terço de gestação, após anestesia geral. A porção dorsal do âmnio foi separada mecanicamente, lavada com PBS e submetida à digestão com colagenase. As células coletadas foram propagadas em cultivo (DMEN-F12/-MEM) e criopreservadas em várias passagens enquanto se efetuava a avaliação da cinética de crescimento e das características morfológicas. Em cultivo, as AMSCs demonstraram aderência à placa e uma morfologia similar a dos fibroblastos.
The amnion derived mesenchymal stem cells (AMSCs) are multipotent cells with a high ability to differentiate into multiple lineages. They can be obtained by non-invasive methods and therefore are exempt from the normal ethical problems involving stem cell use. The aim of this study was to isolate and characterize the progenitor mesenchymal cells from the cat amniotic membrane for future application in cell therapy. The cells were isolated from four fetal membranes collected after a routine ovarian hysterectomy process from cats in their third gestational trimester, under general anesthesia. The dorsal portion of amnion was mechanically separated, washed with PBS and subjected to collagenase digestion. The isolated cells were propagated in culture media (DMEMF12 or -MEM) and frozen in various passages while the growing kinetics and cell morphology were analyzed. In culture medium, AMSCs were adherent to the plastic culture dish and had a morphology similar to fibroblasts.
Assuntos
Animais , Gatos , Gatos/genética , Mesoderma/anatomia & histologia , Mesoderma/embriologia , Separação Celular/classificação , Separação Celular/veterinária , Âmnio/crescimento & desenvolvimento , Citometria de Fluxo , Crescimento Celular , Criopreservação , Imuno-Histoquímica , Imuno-Histoquímica/veterináriaRESUMO
The morphology of the gastrocoel roof plate and the presence of cilia in this structure were examined in embryos of four species of frogs. Embryos of Ceratophrys stolzmanni (Ceratophryidae) and Engystomops randi (Leiuperidae) develop rapidly, provide comparison for the analysis of gastrocoel roof plate development in the slow-developing embryos of Epipedobates machalilla (Dendrobatidae) and Gastrotheca riobambae (Hemiphractidae). Embryos of the analyzed frogs develop from eggs of different sizes, and display different reproductive and developmental strategies. In particular, dorsal convergence and extension and archenteron elongation begin during gastrulation in embryos of rapidly developing frogs, as in Xenopus laevis. In contrast, cells that involute during gastrulation are stored in the large circumblastoporal collar that develops around the closed blastopore in embryos of slow-developing frogs. Dorsal convergence and extension only start after blastopore closure in slow-developing frog embryos. However, in the neurulae, a gastrocoel roof plate develops, despite the accumulation of superficial mesodermal cells in the circumblastoporal collar. Embryos of all four species develop a ciliated gastrocoel roof plate at the beginning of neurulation. Accordingly, fluid-flow across the gastrocoel roof plate is likely the mechanism of left-right asymmetry patterning in these frogs, as in X. laevis and other vertebrates. A ciliated gastrocoel roof plate, with a likely origin as superficial mesoderm, is conserved in frogs belonging to four different families and with different modes of gastrulation.
Assuntos
Cílios/química , Gástrula/embriologia , Ranidae/embriologia , Animais , Embrião não Mamífero , Gástrula/citologia , Gastrulação , Mesoderma/embriologia , Modelos Biológicos , Ranidae/classificação , Ranidae/crescimento & desenvolvimento , Especificidade da EspécieRESUMO
Endocan is a dermatan sulfate proteoglycan (DSPG) that has been observed in the cytoplasm of endothelial cells of small and large vessels in lung, kidney, liver, colon, ovary and brain tumors. This DSPG has been implicated in the regulation of cellular activities such as adhesion, migration, and proliferation. Given the important roles played by endocan in such processes, we sought to determine whether this DSPG is present in the chicken embryo aortic wall in embryonic days 12 and 14, when intimal thickening and endothelial transformation are notorious. Immunolabeling of serial paraffin cross-sections revealed endocan immunoreactivity at the endothelium and some mesenchymal cells constituting the intimal thickening but not in the cells arranged in lamellar layers. We also investigated whether endocan was present in monolayers of primary embryonic aortic endothelial cells attached to fibronectin when they were deprived of serum and stimulated with epidermal growth factor. Immunofluorescence determined that in the epidermal growth factor (EGF) condition where separating, detaching, and migrating cells were observed, endocan appeared organized in arrays typical of focal complexes in the leading edge of these cells. In serum-free medium condition in which the endothelial cells displayed a cobblestone appearance, endocan appeared mainly delineating the margin of many cells. This study demonstrates for the first time the presence of endocan during the aortic wall remodeling, and provides evidence that suggests a possible contribution of this DSPG in the endothelial-mesenchymal transition (EndoMT) process.