RESUMEN
Tissue morphogenesis is intimately linked to the changes in shape and organisation of individual cells. In curved epithelia, cells can intercalate along their own apicobasal axes, adopting a shape named 'scutoid' that allows energy minimization in the tissue. Although several geometric and biophysical factors have been associated with this 3D reorganisation, the dynamic changes underlying scutoid formation in 3D epithelial packing remain poorly understood. Here, we use live imaging of the sea star embryo coupled with deep learning-based segmentation to dissect the relative contributions of cell density, tissue compaction and cell proliferation on epithelial architecture. We find that tissue compaction, which naturally occurs in the embryo, is necessary for the appearance of scutoids. Physical compression experiments identify cell density as the factor promoting scutoid formation at a global level. Finally, the comparison of the developing embryo with computational models indicates that the increase in the proportion of scutoids is directly associated with cell divisions. Our results suggest that apico-basal intercalations appearing immediately after mitosis may help accommodate the new cells within the tissue. We propose that proliferation in a compact epithelium induces 3D cell rearrangements during development.
Asunto(s)
Proliferación Celular , Embrión no Mamífero , Morfogénesis , Animales , Epitelio , Embrión no Mamífero/citología , Recuento de Células , Estrellas de Mar/embriología , Células Epiteliales/citología , Células Epiteliales/metabolismo , División CelularRESUMEN
Primordial germ cells (PGCs) are specified by diverse mechanisms in early development. In some animals, PGCs are specified via inheritance of maternal determinants, while in others, in a process thought to represent the ancestral mode, PGC fate is induced by cell interactions. Although the terminal factors expressed in specified germ cells are widely conserved, the mechanisms by which these factors are regulated can be widely diverse. Here we show that a post-translational mechanism of germ cell specification is conserved between two echinoderm species thought to employ divergent germ line segregation strategies. Sea urchins segregate their germ line early by an inherited mechanism. The DEAD-box RNA - helicase Vasa, a conserved germline factor, becomes enriched in the PGCs by degradation in future somatic cells by the E3-ubiquitin-ligase Gustavus (Gustafson et al., 2011). This post-translational activity occurs early in development, substantially prior to gastrulation. Here we test this process in germ cell specification of sea star embryos, which use inductive signaling mechanisms after gastrulation for PGC fate determination. We find that Vasa-GFP protein becomes restricted to the PGCs in the sea star even though the injected mRNA is present throughout the embryo. Gustavus depletion, however, results in uniform accumulation of the protein. These data demonstrate that Gustavus-mediated Vasa turnover in somatic cells is conserved between species with otherwise divergent PGC specification mechanisms. Since Gustavus was originally identified in Drosophila melanogaster to have similar functions in Vasa regulation (Kugler et al., 2010), we conclude that this node of Vasa regulation in PGC formation is ancestral and evolutionarily transposable from the ancestral, induced PGC specification program to an inherited PGC specification mechanism.
Asunto(s)
ARN Helicasas DEAD-box/metabolismo , Células Germinativas/citología , Erizos de Mar/embriología , Estrellas de Mar/embriología , Ubiquitina-Proteína Ligasas/metabolismo , Animales , Sistemas CRISPR-Cas/genética , Técnicas de Cultivo de Embriones , Embrión no Mamífero/embriología , Procesamiento Proteico-PostraduccionalRESUMEN
Regeneration as an adult developmental process is in many aspects similar to embryonic development. Although many studies point out similarities and differences, no large-scale, direct and functional comparative analyses between development and regeneration of a specific cell type or structure in one animal exist. Here, we use the brittle star Amphiura filiformis to characterise the role of the FGF signalling pathway during skeletal development in embryos and arm regeneration. In both processes, we find ligands expressed in ectodermal cells that flank underlying skeletal mesenchymal cells, which express the receptors. Perturbation of FGF signalling showed inhibited skeleton formation in both embryogenesis and regeneration, without affecting other key developmental processes. Differential transcriptome analysis finds mostly differentiation genes rather than transcription factors to be downregulated in both contexts. Moreover, comparative gene analysis allowed us to discover brittle star-specific differentiation genes. In conclusion, our results show that the FGF pathway is crucial for skeletogenesis in the brittle star, as in other deuterostomes, and provide evidence for the re-deployment of a developmental gene regulatory module during regeneration.
Asunto(s)
Desarrollo Óseo/fisiología , Regeneración Ósea/fisiología , Huesos/embriología , Factores de Crecimiento de Fibroblastos/metabolismo , Estrellas de Mar/embriología , Animales , Huesos/metabolismo , Ectodermo/citología , Ectodermo/metabolismo , Desarrollo Embrionario/genética , Extremidades/crecimiento & desarrollo , Mesodermo/citología , Mesodermo/metabolismo , Pirroles/farmacología , Receptor Tipo 1 de Factor de Crecimiento de Fibroblastos/antagonistas & inhibidores , Transducción de Señal/fisiología , Estrellas de Mar/genética , Estrellas de Mar/metabolismo , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
Biomolecular assemblies govern the physiology of cells. Their function often depends on the changes in molecular arrangements of constituents, both in the positions and orientations. While recent advancements of fluorescence microscopy including super-resolution microscopy have enabled us to determine the positions of fluorophores with unprecedented accuracy, monitoring the orientation of fluorescently labeled molecules within living cells in real time is challenging. Fluorescence polarization microscopy (FPM) reports the orientation of emission dipoles and is therefore a promising solution. For imaging with FPM, target proteins need labeling with fluorescent probes in a sterically constrained manner, but because of difficulties in the rational three-dimensional design of protein connection, a universal method for constrained tagging with fluorophore was not available. Here, we report POLArIS, a genetically encoded and versatile probe for molecular orientation imaging. Instead of using a direct tagging approach, we used a recombinant binder connected to a fluorescent protein in a sterically constrained manner that can target specific biomolecules of interest by combining with phage display screening. As an initial test case, we developed POLArISact, which specifically binds to F-actin in living cells. We confirmed that the orientation of F-actin can be monitored by observing cells expressing POLArISact with FPM. In living starfish early embryos expressing POLArISact, we found actin filaments radially extending from centrosomes in association with microtubule asters during mitosis. By taking advantage of the genetically encoded nature, POLArIS can be used in a variety of living specimens, including whole bodies of developing embryos and animals, and also be expressed in a cell type/tissue specific manner.
Asunto(s)
Citoesqueleto de Actina/metabolismo , Polarización de Fluorescencia/métodos , Colorantes Fluorescentes/metabolismo , Microscopía Fluorescente/métodos , Microtúbulos/metabolismo , Imagen Molecular/métodos , Estrellas de Mar/embriología , Animales , Embrión no Mamífero/citología , Embrión no Mamífero/metabolismo , Proteínas Fluorescentes Verdes/metabolismo , Células HeLa , Humanos , Células LLC-PK1 , PorcinosRESUMEN
The extensive array of morphological diversity among animal taxa represents the product of millions of years of evolution. Morphology is the output of development, therefore phenotypic evolution arises from changes to the topology of the gene regulatory networks (GRNs) that control the highly coordinated process of embryogenesis. A particular challenge in understanding the origins of animal diversity lies in determining how GRNs incorporate novelty while preserving the overall stability of the network, and hence, embryonic viability. Here we assemble a comprehensive GRN for endomesoderm specification in the sea star from zygote through gastrulation that corresponds to the GRN for sea urchin development of equivalent territories and stages. Comparison of the GRNs identifies how novelty is incorporated in early development. We show how the GRN is resilient to the introduction of a transcription factor, pmar1, the inclusion of which leads to a switch between two stable modes of Delta-Notch signaling. Signaling pathways can function in multiple modes and we propose that GRN changes that lead to switches between modes may be a common evolutionary mechanism for changes in embryogenesis. Our data additionally proposes a model in which evolutionarily conserved network motifs, or kernels, may function throughout development to stabilize these signaling transitions.
Asunto(s)
Embrión no Mamífero/metabolismo , Regulación del Desarrollo de la Expresión Génica , Redes Reguladoras de Genes , Erizos de Mar/genética , Estrellas de Mar/genética , Animales , Embrión no Mamífero/embriología , Evolución Molecular , Gastrulación/genética , Mesodermo/embriología , Mesodermo/metabolismo , Modelos Genéticos , Erizos de Mar/embriología , Especificidad de la Especie , Estrellas de Mar/embriología , Factores de Transcripción/genéticaRESUMEN
Understanding how changes in developmental gene expression alter morphogenesis is a fundamental problem in development and evolution. A promising approach to address this problem is to compare the developmental transcriptomes between related species. The echinoderm phylum consists of several model species that have significantly contributed to the understanding of gene regulation and evolution. Particularly, the regulatory networks of the sea star, Patiria miniata (P. miniata), have been extensively studied, however developmental transcriptomes for this species were lacking. Here we generated developmental transcriptomes of P. miniata and compared these with those of two sea urchins species. We demonstrate that the conservation of gene expression depends on gene function, cell type and evolutionary distance. With increasing evolutionary distance the interspecies correlations in gene expression decreases. The reduction is more severe in the correlations between morphologically equivalent stages (diagonal elements) than in the correlation between morphologically distinct stages (off-diagonal elements). This could reflect a decrease in the morphological constraints compared to other constraints that shape gene expression at large evolutionary divergence. Within this trend, the interspecies correlations of developmental control genes maintain their diagonality at large evolutionary distance, and peak at the onset of gastrulation, supporting the hourglass model of phylotypic stage conservation.
Asunto(s)
Evolución Molecular , Perfilación de la Expresión Génica , Regulación del Desarrollo de la Expresión Génica , Estrellas de Mar/crecimiento & desarrollo , Estrellas de Mar/genética , Animales , Homología de Secuencia de Ácido Nucleico , Estrellas de Mar/embriologíaRESUMEN
Specification of the germ cell lineage is required for sexual reproduction in animals. The mechanism of germ cell specification varies among animals but roughly clusters into either inherited or inductive mechanisms. The inductive mechanism, the use of cell-cell interactions for germ cell specification, appears to be the ancestral mechanism in animal phylogeny, yet the pathways responsible for this process are only recently surfacing. Here, we show that germ cell factors in the sea star initially are present broadly, then become restricted dorsally and then in the left side of the embryo where the germ cells form a posterior enterocoel. We find that Nodal signaling is required for the restriction of two germ cell factors, Nanos and Vasa, during the early development of this animal. We learned that Nodal inhibits germ cell factor accumulation in three ways including: inhibition of specific transcription, degradation of specific mRNAs and inhibition of tissue morphogenesis. These results document a signaling mechanism required for the sequential restriction of germ cell factors, which causes a specific set of embryonic cells to become the primordial germ cells.
Asunto(s)
Proteína Nodal/metabolismo , Estrellas de Mar/embriología , Animales , Apoptosis , Linaje de la Célula , ARN Helicasas DEAD-box/metabolismo , Evolución Molecular , Regulación del Desarrollo de la Expresión Génica , Células Germinativas/metabolismo , Modelos Biológicos , ARN Mensajero/genética , ARN Mensajero/metabolismo , Proteínas de Unión al ARN/metabolismo , Transducción de Señal , Estrellas de Mar/genética , Estrellas de Mar/metabolismoRESUMEN
In this study, we attempted to reveal fundamental aspects of starfish embryogenesis, particularly embryonic axis specification or determination, in Patiria pectinifera. We first cloned PpNodal, which is known to play an important role in the specification of the embryonic axis in a wide range of animals, and studied its expression profile. PpNodal expression was first detected at the mid-blastula stage and showed a single peak around the onset of gastrulation. These features of Nodal expression were shifted to later stages by several hours, compared with those of sea urchin embryos. After the gastrulation started, the expression level became gradually lowered up to the early bipinnaria stage, while the expression level became drastically lowered in sea urchin embryos during gastrulation. The localized Nodal expression in the presumptive oral region was not observed in starfish embryos, unlike in sea urchin embryos. Furthermore, SB431542, an inhibitor of Nodal receptor, did not affect the formation of the DV axis, although it caused the loss of left-right asymmetry. In contrast to this, SB525334, a specific inhibitor of TGF-beta receptor, caused the complete loss of the DV axis. Thus, the usage of signaling molecules during early embryogenesis likely varies among echinoderm classes.
Asunto(s)
Tipificación del Cuerpo/fisiología , Estrellas de Mar/embriología , Estrellas de Mar/metabolismo , Animales , Benzamidas/farmacología , Tipificación del Cuerpo/genética , Dioxoles/farmacología , Regulación del Desarrollo de la Expresión Génica/genética , Regulación del Desarrollo de la Expresión Génica/fisiología , Proteína Nodal/genética , Proteína Nodal/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genética , Transducción de Señal/fisiologíaRESUMEN
It has long been argued that heterochrony, a change in relative timing of a developmental process, is a major source of evolutionary innovation. Heterochronic changes of regulatory gene activation could be the underlying molecular mechanism driving heterochronic changes through evolution. Here, we compare the temporal expression profiles of key regulatory circuits between sea urchin and sea star, representative of two classes of Echinoderms that shared a common ancestor about 500 million years ago. The morphologies of the sea urchin and sea star embryos are largely comparable, yet, differences in certain mesodermal cell types and ectodermal patterning result in distinct larval body plans. We generated high resolution temporal profiles of 17 mesodermally-, endodermally- and ectodermally-expressed regulatory genes in the sea star, Patiria miniata, and compared these to their orthologs in the Mediterranean sea urchin, Paracentrotus lividus. We found that the maternal to zygotic transition is delayed in the sea star compared to the sea urchin, in agreement with the longer cleavage stage in the sea star. Interestingly, the order of gene activation shows the highest variation in the relatively diverged mesodermal circuit, while the correlations of expression dynamics are the highest in the strongly conserved endodermal circuit. We detected loose scaling of the developmental rates of these species and observed interspecies heterochronies within all studied regulatory circuits. Thus, after 500 million years of parallel evolution, mild heterochronies between the species are frequently observed and the tight temporal scaling observed for closely related species no longer holds.
Asunto(s)
Regulación del Desarrollo de la Expresión Génica , Erizos de Mar/embriología , Estrellas de Mar/embriología , Animales , Tipificación del Cuerpo , Ectodermo , Evolución Molecular , Perfilación de la Expresión Génica , Mesodermo/metabolismo , Reacción en Cadena de la Polimerasa , ARN/metabolismo , Especificidad de la Especie , CigotoRESUMEN
Reconstruction of a starfish embryo provides unique morphogenesis during the developmental process that is not observed in normal development. Here, we established a novel method for reconstruction from single embryos/larvae. By using this method, we investigated the morphogenetic capabilities in critical steps during the reconstruction process as showed by the reconstructed embryos generated from embryos/larvae at the six developmental stages, or from segregated ectodermal and/or endomesodermal cells. Additionally, the novel method addressed several problems found in prior methods related to reproducibly generating reconstructed embryos. In the reconstructions from the various stage embryos/larvae, the morphogenetic capabilities were substantively reduced in the reconstructed embryos generated from 3-day bipinnaria (3dBp). The combination experiments using ectodermal or endomesodermal cells segregated from 2dBp or 3dBp showed a reduction of the morphogenetic capabilities in both cells types in 3dBp. The reconstructed embryos generated from ectodermal or endomesodermal cells segregated from 2dBp possessed partial morphological features, such as formation of the epithelium or blastopore, but all failed to develop into bipinnariae. These results indicate two limitations of the morphogenetic capabilities during the reconstruction process. Firstly, the morphogenetic capabilities to reconstruct an embryo are considerably reduced between 2dBp and 3dBp. Secondly, cells specified as ectoderm or endomesoderm possess limited morphogenetic capabilities to reconstruct bipinnaria. Furthermore, our results demonstrate that the interaction between these specified cell types is required for reconstruction.
Asunto(s)
Embrión no Mamífero/embriología , Estrellas de Mar/embriología , Animales , Ectodermo/citología , Ectodermo/metabolismo , Embrión no Mamífero/metabolismo , Larva/citología , Larva/metabolismo , Mesodermo/citología , Mesodermo/metabolismo , Estrellas de Mar/metabolismoRESUMEN
How neural stem cells generate the correct number and type of differentiated neurons in appropriate places remains an important question. Although nervous systems are diverse across phyla, in many taxa the larva forms an anterior concentration of serotonergic neurons, or apical organ. The sea star embryo initially has a pan-neurogenic ectoderm, but the genetic mechanism that directs a subset of these cells to generate serotonergic neurons in a particular location is unresolved. We show that neurogenesis in sea star larvae begins with soxc-expressing multipotent progenitors. These give rise to restricted progenitors that express lhx2/9 soxc- and lhx2/9-expressing cells can undergo both asymmetric divisions, allowing for progression towards a particular neural fate, and symmetric proliferative divisions. We show that nested concentric domains of gene expression along the anterior-posterior (AP) axis, which are observed in a great diversity of metazoans, control neurogenesis in the sea star larva by promoting particular division modes and progression towards becoming a neuron. This work explains how spatial patterning in the ectoderm controls progression of neurogenesis in addition to providing spatial cues for neuron location. Modification to the sizes of these AP territories provides a simple mechanism to explain the diversity of neuron number among apical organs.
Asunto(s)
Tipificación del Cuerpo/genética , Redes Reguladoras de Genes , Sistema Nervioso/embriología , Neurogénesis/genética , Estrellas de Mar/embriología , Estrellas de Mar/genética , Animales , Animales Modificados Genéticamente , Diferenciación Celular/genética , Células Cultivadas , Ectodermo/embriología , Ectodermo/metabolismo , Embrión no Mamífero , Regulación del Desarrollo de la Expresión Génica , Sistema Nervioso/metabolismo , Neuronas Serotoninérgicas/fisiologíaRESUMEN
Nanos is a translational regulator required for the survival and maintenance of primordial germ cells. In the sea urchin, Strongylocentrotus purpuratus (Sp), Nanos 2 mRNA is broadly transcribed but accumulates specifically in the small micromere (sMic) lineage, in part because of the 3'UTR element GNARLE leads to turnover in somatic cells but retention in the sMics. Here we found that the Nanos 2 protein is also selectively stabilized; it is initially translated throughout the embryo but turned over in the future somatic cells and retained only in the sMics, the future germ line in this animal. This differential stability of Nanos protein is dependent on the open reading frame (ORF), and is independent of the sumoylation and ubiquitylation pathways. Manipulation of the ORF indicates that 68 amino acids in the N terminus of the Nanos protein are essential for its stability in the sMics whereas a 45 amino acid element adjacent to the zinc fingers targets its degradation. Further, this regulation of Nanos protein is cell autonomous, following formation of the germ line. These results are paradigmatic for the unique presence of Nanos in the germ line by a combination of selective RNA retention, distinctive translational control mechanisms (Oulhen et al., 2013), and now also by defined Nanos protein stability.
Asunto(s)
Células Germinativas/metabolismo , Sistemas de Lectura Abierta/genética , Proteínas de Unión al ARN/genética , Proteínas de Unión al ARN/metabolismo , Strongylocentrotus purpuratus/embriología , Animales , Regulación del Desarrollo de la Expresión Génica , Técnicas de Inactivación de Genes , Morfolinos , Biosíntesis de Proteínas/genética , Estabilidad Proteica , ARN Mensajero/biosíntesis , Estrellas de Mar/embriología , Sumoilación , UbiquitinaciónRESUMEN
The provision of yolk precursor proteins to the oviparous egg is crucial for normal embryo development. In Echinodermata, a transferrin-like yolk component termed major yolk protein (MYP) is a major precursor protein in Echinoidea and Holothuroidea. In contrast, in Asteroidea a single vitellogenin (Vtg) was recently identified, but its role as primary yolk protein remains unclear. To resolve the apparent MYP-Vtg dichotomy in sea stars and to understand the dynamics of candidate yolk protein gene expression during the reproductive cycle, we investigated the molecular structures of sea star Vtg and MYP and quantified their transcript levels during oogenesis. By combining protein sequencing of the predominant proteins in ovulated eggs of Patiriella regularis with ovarian transcriptome sequencing and molecular cloning, we characterized two cDNAs encoding two bona fide Vtgs (PrVtg1 and PrVtg2) and a partial cDNA encoding MYP (PrMYP). PrMYP mRNA was found in low abundance in growing oocytes, possibly as maternal transcripts for translation after ovulation. In contrast, PrVtg transcripts, whose levels varied during the reproductive cycle, were not found in developing oocytes - rather, they were detected in ovarian follicle cells and pyloric caeca, indicating an extra-oocytic origin. Vtg accumulating in oocytes was stored in the form of cleaved products, which constituted the most abundant yolk polypeptides in ovulated sea star eggs; their levels decreased during early embryonic and larval development. Together, these traits are the hallmarks of a classical yolk protein - and hence, we contend that Vtg, and not MYP, is the main yolk protein in asteroids.
Asunto(s)
Proteínas del Huevo/genética , Estrellas de Mar/genética , Vitelogeninas/genética , Secuencia de Aminoácidos , Animales , Clonación Molecular , Secuencia Conservada , Proteínas del Huevo/metabolismo , Embrión no Mamífero , Femenino , Expresión Génica , Oocitos/fisiología , Ovario/fisiología , Filogenia , Pepinos de Mar/genética , Erizos de Mar/genética , Estrellas de Mar/embriología , Vitelogeninas/metabolismoRESUMEN
Echinoderm pluteus larvae are considered a classical example of convergent evolution that occurred in sea urchins and brittle stars. Several genes are known to be involved in the development of pluteus arms in sea urchins, including fgfA, pax2/5/8, pea3, otp, wnt5, and tet. To determine whether the convergent evolution of larval arms also involves these genes in brittle stars, their expression patterns were determined in brittle star. We found that all genes showed similar expression in the arms of ophiopluteus to that seen in echinopluteus, suggesting that convergent evolution of pluteus arms occurred by recruitment of a similar set of genes. This may be explained by our observation that some of these genes are also expressed in the spine rudiment of direct-type development sea urchins. We propose an evolutionary scenario wherein the pluteus arms of both echinopluteus and ophiopluteus were acquired by independent co-options of the genetic module responsible for the projection of the adult skeleton.
Asunto(s)
Erizos de Mar/anatomía & histología , Erizos de Mar/genética , Estrellas de Mar/anatomía & histología , Estrellas de Mar/genética , Animales , Erizos de Mar/embriología , Estrellas de Mar/embriología , TranscriptomaRESUMEN
In many invertebrates, the nuclearization of ß-catenin at one pole of the embryo initiates endomesoderm specification. An intriguing possibility is that a gradient of nuclear ß-catenin (nß-catenin), similar to that operating in vertebrate neural tube patterning, functions to distinguish cell fates in invertebrates. To test this hypothesis, we determined the function of nß-catenin during the early development of the sea star, which undergoes a basal deuterostomal mode of embryogenesis. We show that low levels of nß-catenin activity initiate bra, which is expressed in the future posterior endoderm-fated territory; intermediate levels are required for expression of foxa and gata4/5/6, which are later restricted to the endoderm; and activation of ets1 and erg in the mesoderm-fated territory requires the highest nß-catenin activity. Transcription factors acting downstream of high nß-catenin segregate the endoderm/mesoderm boundary, which is further reinforced by Delta/Notch signaling. Significantly, therefore, in sea stars, endomesoderm segregation arises through transcriptional responses to levels of nß-catenin activity. Here, we describe the first empirical evidence of a dose-dependent response to a dynamic spatiotemporal nß-catenin activity that patterns cell fates along the primary axis in an invertebrate.
Asunto(s)
Tipificación del Cuerpo , Núcleo Celular/metabolismo , Endodermo/embriología , Mesodermo/embriología , beta Catenina/metabolismo , Animales , Blástula/efectos de los fármacos , Blástula/metabolismo , Tipificación del Cuerpo/efectos de los fármacos , Tipificación del Cuerpo/genética , Núcleo Celular/efectos de los fármacos , Embrión no Mamífero/efectos de los fármacos , Embrión no Mamífero/metabolismo , Endodermo/efectos de los fármacos , Endodermo/metabolismo , Regulación del Desarrollo de la Expresión Génica , Redes Reguladoras de Genes , Factor Nuclear 3-gamma del Hepatocito/metabolismo , Cloruro de Litio/farmacología , Mesodermo/efectos de los fármacos , Mesodermo/metabolismo , Modelos Biológicos , Receptores Notch/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genética , Estrellas de Mar/embriología , Estrellas de Mar/genética , Factores de Tiempo , Transcripción Genética/efectos de los fármacosRESUMEN
BACKGROUND: Echinodermata is a diverse phylum, a sister group to chordates, and contains diverse organisms that may be useful to understand varied mechanisms of germ-line specification. RESULTS: We tested 23 genes in development of the sea star Patiria miniata that fall into five categories: (1) Conserved germ-line factors; (2) Genes involved in the inductive mechanism of germ-line specification; (3) Germ-line associated genes; (4) Molecules involved in left-right asymmetry; and (5) Genes involved in regulation and maintenance of the genome during early embryogenesis. Overall, our results support the contention that the posterior enterocoel is a source of the germ line in the sea star P. miniata. CONCLUSIONS: The germ line in this organism appears to be specified late in embryogenesis, and in a pattern more consistent with inductive interactions amongst cells. This is distinct from the mechanism seen in sea urchins, a close relative of the sea star clad. We propose that P. miniata may serve as a valuable model to study inductive mechanisms of germ-cell specification and when compared with germ-line formation in the sea urchin S. purpuratus may reveal developmental transitions that occur in the evolution of inherited and inductive mechanisms of germ-line specification.
Asunto(s)
Embrión no Mamífero , Células Germinativas , Modelos Biológicos , Erizos de Mar , Estrellas de Mar , Animales , Embrión no Mamífero/citología , Embrión no Mamífero/embriología , Células Germinativas/citología , Células Germinativas/metabolismo , Erizos de Mar/citología , Erizos de Mar/embriología , Especificidad de la Especie , Estrellas de Mar/citología , Estrellas de Mar/embriologíaRESUMEN
The Wnt signaling pathway is highly conserved across metazoa and has pleiotropic functions in the development of many animals. Binding of a secreted Wnt ligand to its Frizzled (Fz) receptor activates Dishevelled, which then drives one of three major signaling cascades, canonical (ß-catenin), calcium, or planar cell polarity signaling. These pathways have distinct developmental effects and function in different processes in different organisms. Here we report the expression of six wnt and three fz genes during embryogenesis of the sea star, Patiria miniata, as a first step in uncovering the roles of Wnt signaling in the development of this organism. wnt3, wnt4, wnt8, and wnt16 are expressed in nested domains in the endoderm and lateral ectoderm from blastula through late gastrula stages; wnt2 and wnt5 are expressed in the mesoderm and anterior endoderm. Expression of different fz paralogs is detected in the mesoderm; posterior endoderm and ectoderm; and anterior ectoderm. Taken together, this suggests that Wnt signaling can occur throughout most of the embryo and may therefore play multiple roles during sea star development.
Asunto(s)
Receptores Frizzled/genética , Expresión Génica , Estrellas de Mar/genética , Proteínas Wnt/genética , Secuencia de Aminoácidos , Animales , Blástula/metabolismo , Receptores Frizzled/química , Receptores Frizzled/metabolismo , Gástrula/metabolismo , Regulación del Desarrollo de la Expresión Génica , Datos de Secuencia Molecular , Filogenia , Estrellas de Mar/embriología , Estrellas de Mar/metabolismo , Proteínas Wnt/química , Proteínas Wnt/metabolismoRESUMEN
BACKGROUND: The ParaHox genes are thought to be major players in patterning the gut of several bilaterian taxa. Though this is a fundamental role that these transcription factors play, their activities are not limited to the endoderm and extend to both ectodermal and mesodermal tissues. Three genes compose the ParaHox group: Gsx, Xlox and Cdx. In some taxa (mostly chordates but to some degree also in protostomes) the three genes are arranged into a genomic cluster, in a similar fashion to what has been shown for the better-known Hox genes. Sea urchins possess the full complement of ParaHox genes but they are all dispersed throughout the genome, an arrangement that, perhaps, represented the primitive condition for all echinoderms. In order to understand the evolutionary history of this group of genes we cloned and characterized all ParaHox genes, studied their expression patterns and identified their genomic loci in a member of an earlier branching group of echinoderms, the asteroid Patiria miniata. RESULTS: We identified the three ParaHox orthologs in the genome of P. miniata. While one of them, PmGsx is provided as maternal message, with no zygotic activation afterwards, the other two, PmLox and PmCdx are expressed during embryogenesis, within restricted domains of both endoderm and ectoderm. Screening of a Patiria bacterial artificial chromosome (BAC) library led to the identification of a clone containing the three genes. The transcriptional directions of PmGsx and PmLox are opposed to that of the PmCdx gene within the cluster. CONCLUSIONS: The identification of P. miniata ParaHox genes has revealed the fact that these genes are clustered in the genome, in contrast to what has been reported for echinoids. Since the presence of an intact cluster, or at least a partial cluster, has been reported in chordates and polychaetes respectively, it becomes clear that within echinoderms, sea urchins have modified the original bilaterian arrangement. Moreover, the sea star ParaHox domains of expression show chordate-like features not found in the sea urchin, confirming that the dynamics of gene expression for the respective genes and their putative regulatory interactions have clearly changed over evolutionary time within the echinoid lineage.
Asunto(s)
Cordados/genética , Genes Homeobox/genética , Proteínas de Homeodominio/genética , Familia de Multigenes/genética , Estrellas de Mar/genética , Secuencia de Aminoácidos , Animales , Ectodermo/embriología , Ectodermo/metabolismo , Desarrollo Embrionario/genética , Evolución Molecular , Tracto Gastrointestinal/embriología , Tracto Gastrointestinal/metabolismo , Perfilación de la Expresión Génica , Regulación del Desarrollo de la Expresión Génica , Genoma/genética , Proteínas de Homeodominio/química , Proteínas de Homeodominio/metabolismo , Datos de Secuencia Molecular , Homología de Secuencia de Aminoácido , Estrellas de Mar/embriología , Factores de TiempoRESUMEN
A great challenge in development biology is to understand how interacting networks of regulatory genes can direct the often highly complex patterning of cells in a 3D embryo. Here, we detail the gene regulatory network that describes the distribution of ciliary band-associated neurons in the bipinnaria larva of the sea star. This larva, typically for the ancestral deuterostome dipleurula larval type that it represents, forms two loops of ciliary bands that extend across much of the anterior-posterior and dorsal-ventral ectoderm. We show that the sea star first likely uses maternally inherited factors and the Wnt and Delta pathways to distinguish neurogenic ectoderm from endomesoderm. The broad neurogenic potential of the ectoderm persists throughout much of gastrulation. Nodal, bone morphogenetic protein 2/4 (Bmp2/4), and Six3-dependent pathways then sculpt a complex ciliary band territory that is defined by the expression of the forkhead transcription factor, foxg. Foxg is needed to define two molecularly distinct ectodermal domains, and for the formation of differentiated neurons along the edge of these two territories. Thus, significantly, Bmp2/4 signaling in sea stars does not distinguish differentiated neurons from nonneuronal ectoderm as it does in many other animals, but instead contributes to the patterning of an ectodermal territory, which then, in turn, provides cues to permit the final steps of neuronal differentiation. The modularity between specification and patterning likely reflects the evolutionary history of this gene regulatory network, in which an ancient module for specification of a broad neurogenic potential ectoderm was subsequently overlaid with a module for patterning.
Asunto(s)
Tipificación del Cuerpo/fisiología , Embrión no Mamífero/embriología , Neurogénesis/fisiología , Estrellas de Mar/embriología , Vía de Señalización Wnt/fisiología , Animales , Secuencia de Bases , Proteína Morfogenética Ósea 2/metabolismo , Proteína Morfogenética Ósea 4/metabolismo , Ectodermo/citología , Ectodermo/embriología , Embrión no Mamífero/citología , Endodermo/citología , Endodermo/embriología , Gastrulación/fisiología , Regulación del Desarrollo de la Expresión Génica/fisiología , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Proteínas de la Membrana/metabolismo , Datos de Secuencia Molecular , Estrellas de Mar/citología , Proteínas Wnt/metabolismoRESUMEN
A new C30 linear polyacetylene compound designated petroacetylene (1) has been isolated from the marine sponge Petrosia solida Hoshino 1981, collected off the coast of Amami-Oshima, Kagoshima Prefecture, Japan. The structure was elucidated on the basis of spectroscopic data and chemical means. Petroacetylene (1) inhibited blastulation of starfish embryos at a concentration of 3.1 µg mL(- 1) or greater.