pinecone/core-2020-05-10-deduplication huggingface dataset:
Dorsal Determinants in theXenopusEgg Are Firmly Associated with the Vegetal Cortex and Behave like Activators of theWntPathway

AbstractTheXenopusegg contains maternal dorsal determinants that are specifically located at the vegetal cortex. To study physical and functional properties of the dorsal determinants, we took advantage of the animal–vegetal reversed embryo. The animal–vegetal reversed embryo is produced by inversion of the fertilized egg, which results in formation of ectoderm and endoderm from the unpigmented and the pigmented halves, respectively [Neffet al.(1983).Dev. Biol.97, 103–112; Black and Gerhart (1985).Dev. Biol.108, 310–324]. We demonstrated by cytoplasmic transplantation that the dorsal activity was specifically localized to the unpigmented cortical cytoplasm of the inverted egg, which is segregated into the future ectodermal lineage. This result suggests that the dorsal determinants are associated with the unpigmented cortex and are not dislodged by the inversion. In addition, we found that two vegetally localized transcripts,Xcat2andVg1mRNAs, were present in the reversed animal pole of the inverted egg, suggesting their association with the unpigmented cortex. In order to compare the dorsal determinant activity with known dorsalizing molecules, we examined the expression pattern ofXnr3andSiamoisin the reversed embryo because these two genes are activated by theWnt-pathway activators (Xwnt-8,β-catenin,etc.) but not by other dorsalizing molecules (noggin, BVg1,etc.). Animal cap of the reversed embryo, which received the unpigmented cortex of the egg, expressedXnr3andSiamois.However,Mix.1,a marker expressed in endoderm and mesoderm in the normal embryo in response to mesodermal inducers, was not detected in the animal cap of the reversed embryo. In addition, we found that β-cateninprotein accumulated in nuclei of unpigmented animal pole cells of the reversed embryo. These results suggest that the maternal dorsal determinants behave more similarly to theWnt-pathway activators thannogginorBVg1
Cellular Mechanism Underlying Neural Convergent Extension inXenopus laevisEmbryos

AbstractConvergent extension, the simultaneous narrowing and lengthening of a tissue, plays a major role in shaping and patterning the neural ectoderm in vertebrate embryos. In this paper, we characterize the cellular mechanism underlying convergent extension of the neural ectoderm in theXenopus laevislate gastrula and neurula embryo. Neural ectoderm inX. laevisconsists of two components, a superficial layer of epithelial cells overlying deep mesenchymal cells. To investigate the force contribution of the deep cells to convergent extension, we explanted single layers of neural deep cells from late gastrula stage embryos. These “neural deep cell explants” undergo active convergent extension autonomously, implying that these cells contribute force for neural convergent extensionin vivo.Using time-lapse videorecording of these explants, we observed the neural deep cell behaviors (previously hidden behind an opaque epithelium) underlying convergent extension. We show that neural deep cells mediolaterally intercalate to form a longer, narrower tissue and that cell shape change and cell division contribute little to their convergent extension. Moreover, we characterize the neural deep cell motility driving mediolateral intercalation, also using time-lapse videorecordings. Analyses of these videos revealed that, on average, neural deep cells exhibit mediolaterally biased protrusive activity which is expressed in an episodic fashion. We propose that neural deep cells accomplish mediolateral intercalation by applying their protrusions upon one another, exerting traction, and pulling themselves between one another. This mechanism is similar to that previously described for convergent extension of the mesodermal cells. However, because the neural deep cells do not mediolaterally elongate during their convergent extension as the mesodermal cells do, we predict that a given intercalation will result in more extension for neural deep cells than for the mesodermal cells. Intercalation of neural cells also likely occurs in a more episodic manner than that of the mesodermal cells because the neural cells’ mediolateral protrusive activity is episodic, whereas the protrusive activity of mesodermal cells is more continuous. These differences in protrusive activity and cell shape changes between the neural and mesodermal regions may reflect specializations of the same basic mechanism of mediolateral intercalation, tailored to accommodate other aspects of patterning and development of each tissue. These descriptions of the active cell motility underlying neural convergent extension inX. laevisare the first high-resolution video documentation of protrusive activity driving neural convergent extension in any system. Our findings provide an important step in the investigation of neural convergent extension inX. laevisand further our understanding of convergent extension in general
The Exocytosis Regulatory Proteins Syntaxin and VAMP Are Shed from Sea Urchin Sperm during the Acrosome Reaction

AbstractSyntaxin is a cytoplasmically oriented plasma membrane protein and VAMP (vesicle-associated membrane protein; synaptobrevin) is a protein associated with the secretory vesicle membrane. These two proteins form part of a complex which is thought to mediate the fusion of plasma and vesicle membranes during exocytosis. This paper reports the identification of syntaxin and VAMP homologues in sea urchin sperm. During fertilization, sea urchin sperm release the contents of a single vesicle, the acrosomal vesicle, exposing the membrane destined to fuse with the egg. During acrosomal exocytosis, the plasma membrane over the acrosomal vesicle fuses at multiple points with the acrosomal membrane (vesiculation) and syntaxin and VAMP are shed with the resulting membrane vesicles. Sea urchin sperm syntaxin and VAMP are associated in a complex as detected by immunoprecipitation. Following acrosomal exocytosis, syntaxin and VAMP cosediment to denser fractions on sucrose gradients showing that they have undergone associative changes during or after the acrosome reaction. Syntaxin and VAMP localization and loss during acrosomal exocytosis support a role for these proteins in regulating the acrosome reaction
A Role for WNT Proteins in Induction of Dermomyotome

AbstractDorsoventral patterning of somites into sclerotome and dermomyotome involves antagonistic actions of ventralizing and dorsalizing signals originating from tissues surrounding the somites. The notochord and the floor plate of the neural tube provide a ventralizing signal(s) directing sclerotome development, whereas the surface ectoderm and dorsal neural tube provide a dorsalizing signal(s) directing dermomyotome development. Evidence has been provided that Sonic Hedgehog mediates the ventralizing effects of notochord and floor plate, but the dorsalizing signal(s) that patterns the dermomyotome has not been identified. The documented expression ofWnt1andWnt3ain the dorsal neural tube and ofWnt4andWnt6in the surface ectoderm at the time of dermomyotome specification prompted us to investigate the involvement of WNT proteins in patterning the dermomyotome. Here we show that tissue culture cells expressing these WNT family members can maintain and induce dermomyotome marker expression in presomitic mesoderm explants, supporting the hypothesis that WNT proteins mediate the dorsalizing effects of the surface ectoderm and dorsal neural tube on somites
Onset of CNTFRα Expression and Signal Transduction during Neurogenesis in Chick Sensory Dorsal Root Ganglia

AbstractThe expression of ciliary neurotrophic factor receptor α (CNTFRα) was investigated in the developing chick dorsal root ganglion (DRG) using affinity-purified anti-CNTFRα antibodies. At thoracic levels, CNTFRα-immunoreactivity (CNTFRα-IR) was first observed at stage 19 (E3) in cells with neuronal morphology. CNTFRα-IR is restricted to the neuronal lineage in the DRG throughout development. CNTFRα expression precedes that of neuron-specific β tubulin, Hu antigen, and Q211 antigen, which are markers expressed in developing sensory neurons. [3H]Thymidine-labeling studies showed the onset of CNTFRα expression during terminal mitosis of sensory neuron precursors, making CNTFRα the earliest known neuronal marker in the DRG. CNTFRα-mediated signal transduction was demonstrated in E7 and E11 DRG neuron cultures by CNTF-induced STAT3 phosphorylation. Although low ligand concentrations (5 pM) elicit STAT3 phosphorylation in E7 and E11 DRG neurons, a survival response is only observed in neurons from E11 DRG. This implicates a complex readout mechanism downstream of STAT3 phosphorylation leading to different cellular responses that depend on the age of the DRG neuron. These results argue against a role of CNTFRα ligands in the control of early neuron survival but are compatible with other functions in neurogenesis and sensory neuron development
Budhead,a Fork Head/HNF-3 Homologue, Is Expressed during Axis Formation and Head Specification in Hydra

AbstractAccumulating evidence indicates that a common set of genes and mechanisms regulates the developmental processes of a variety of triploblastic organisms despite large variation in their body plans. To what extent these same genes and mechanisms are also conserved among diploblasts, which arose earlier in metazoan evolution, is unclear. We have characterized a hydra homologue of the fork head/HNF-3 class of winged-helix proteins, termedbudhead,whose expression patterns suggest a role(s) similar to that found in vertebrates. The vertebrateHNF-3β homologues are expressed early in embryogenesis in regions that have organizer properties, and later they have several roles, among them an important role in rostral head formation. In the adult hydra, where axial patterning processes are continuously active,budheadis expressed in the upper part of the head, which has organizer properties. It is also expressed during the formation of a new axis as part of the development of a bud, hydra's asexual form of reproduction. Expression during later stages of budding, during head regeneration and the formation of ectopic heads, indicates a role in head formation. It is likely thatbudheadplays a critical role in head as well as axis formation in hydra
Heterogeneous Expression of Multiple Putative Patterning Genes by Single Cells from the Chick Hindbrain

AbstractThe metameric organization of the vertebrate hindbrain into rhombomeres appears to result from the patterned expression of several transcription factors and putative signaling molecules. We have applied a refined single-cell reverse transcription-polymerase chain reaction strategy to examine the molecular logic proposed to pattern the hindbrain at the single-cell level. This technique allows analysis of the concurrent expression of several genes within an individual cell at higher sensitivity than byin situhybridization. Our results demonstrate that cells in rhombomere (r) 4 and r5 are heterogeneous in their expression of Hoxa-3, Hoxb-2, Sek-1, and Krox-20, suggesting that single cells are dynamically regulating their rhombomere-specific gene-expression profiles. Furthermore, the strong correlation between Sek-1 and Krox-20 expression at stage 12 was greatly diminished by stage 16, suggesting that the proposed interdependence of these two genes is present only at early stages of hindbrain development
Mixed Cultures of Avian Blastoderm Cells and the Quail Mesoderm Cell Line QCE-6 Provide Evidence for the Pluripotentiality of Early Mesoderm

AbstractDuring the early stages of embryogenesis, the mesoderm gives rise to cells of the cardiovascular system which include cardiac myocytes and vascular endothelial and red blood cells. We have investigated the development of these cell phenotypes using aggregate cultures of avian blastoderm cells, which replicated mesodermal cell diversification. The cell phenotypes expressed by the blastoderm cells were dependent upon the age of the blastoderm cells, with Hamburger–Hamilton stage 3 or 4 cells giving rise to endothelial and red blood cells and stage 5 cells producing endothelial and myocardial cells. To begin to understand the stage dependency of the cellular diversification of these aggregate cultures, we treated the cultures with various signaling factors that have been shown to be present in the early avian embryo. These experiments showed that stem cell factor and TGFα altered cell phenotypes by stimulating red blood cell and myocardial differentiation, respectively. The ability of these growth factors to shift the differentiation profile of aggregate cultures demonstrated the plasticity of early embryonic cells. To explore the diversification of individual mesodermal cells, labeled QCE-6 cells were incorporated within these blastoderm aggregate cultures. Previous studies have shown that this quail mesodermal cell line possesses characteristics of early nondifferentiated mesodermal cells and can be induced to express either myocardial or endothelial cell phenotypes (C. A. Eisenberg and D. M. Bader, 1996,Circ. Res.78, 205–216). In the present study, we show that when these cells were cultured as a component of blastoderm cell aggregates, they differentiated into fully contractile cardiomyocytes or endothelial or red blood cells. Moreover, QCE-6 cell differentiation was in accordance with that displayed by the blastoderm cells. Specifically, QCE-6 cells differentiated into red blood cells when cultured within stage 3 or stage 4, but not stage 5, blastoderm cell aggregates. Accordingly, the differentiation of QCE-6 cells into beating cardiomyocytes only occurred when these cells were incorporated into stage 5 blastoderm cell aggregates. The identical sorting and differentiation patterns that were exhibited by QCE-6 and blastoderm cells suggest that expression of differentiated cell types within the early mesoderm is directed by the surrounding environment without immediate cellular commitment. In addition, these results provide further evidence that QCE-6 cells are representative of a multipotential mesodermal stem cell and that they possess the potential to exhibit fully differentiated cell phenotypes
Null Mutations of theDictyosteliumCyclic Nucleotide Phosphodiesterase Gene Block Chemotactic Cell Movement in Developing Aggregates

AbstractExtracellular cAMP is a critical messenger in the multicellular development of the cellular slime moldDictyostelium discoideum.The levels of cAMP are controlled by a cyclic nucleotide phosphodiesterase (PDE) that is secreted by the cells. The PDE gene (pdsA) is controlled by three promoters that permit expression during vegetative growth, during aggregation, and in prestalk cells of the older structures. Targeted disruption of the gene aborts development, and complementation with a modifiedpdsArestores development. Two distinct promoters must be used for full complementation, and an inhibitory domain of the PDE must be removed. We took advantage of newly isolated PDE-null cells and the natural chimerism of the organism to ask whether the absence of PDE affected individual cell behavior. PDE-null cells aggregated with isogenic wild-type cells in chimeric mixtures, but could not move in a coordinated manner in mounds. The wild-type cells move inward toward the center of the mound, leaving many of the PDE-null cells at the periphery of the aggregate. During the later stages of development, PDE-null cells in the chimera segregate to regions which correspond to the prestalk region and the rear of the slug. Participation in the prespore/spore population returns with the restoration of a modifiedpdsAto the null cells
Single-Cell Analysis of Regulatory Gene Expression in Quiescent and Activated Mouse Skeletal Muscle Satellite Cells

AbstractRepair and regeneration of adult skeletal muscle are mediated by satellite cells. In healthy muscle these rare mononucleate muscle precursor cells are mitotically quiescent. Upon muscle injury or degeneration, members of this self-renewing pool are activated to proliferate and then differentiate. Here we analyzed in single satellite cells the expression of a set of regulatory genes that are candidates for causal roles in satellite cell activation, maturation, and differentiation. Individual cells were identified as satellite cells and selected for analysis based on their physical association with single explanted myofibers or their position beneath the basal lamina in unperturbed muscle tissue. Using a multiplex single-cell RT-PCR assay we simultaneously monitored expression of all four MyoD family regulators of muscle determination and differentiation (MRFs) together with two candidate markers of satellite cell identity, c-met and m-cadherin. By making these measurements on large numbers of individual cells during the time course of satellite cell activation, we were able to define which expression states (possible combinations of the six genes) were represented and to specify how the representation of each state changed with time. Activated satellite cells began to express either MyoD or myf5 first among the MRFs; most cells then expressed both myf-5 and MyoD simultaneously; myogenin came on later in cells expressing both MyoD and myf5; and many cells ultimately expressed all four MRFs simultaneously. The results for fiber-associated satellite cells from either predominantly fast or slow muscles were indistinguishable from each other. The c-met receptor tyrosine kinase was also monitored because it is a candidate for mediating activation of quiescent satellite cells (Allenet al.,1995) and because it might also be a candidate molecular marker for satellite cells. A significant difficulty in studying mouse satellite cells has been the absence of molecular markers that could identify them in the quiescent state before expression of MRFs or desmin and distinguish them from fibroblasts. We show here that c-met receptor is present beneath the basal lamina on presumptive satellite cells in intact muscle and that c-met mRNA and protein are expressed by all myofiber-associated satellite cells from the time of explant through the course of activation, proliferation, and differentiation. c-met was not detected in muscle-derived fibroblasts or in other mononucleate cells from healthy muscle explants. When compared directly with m-cadherin, which has previously been suggested as a marker for quiescent satellite cells, m-cadherin mRNA was detected only in a small subset of satellite cells at early times after myofiber explant. However, at late times following activation (by 96 hr in this fiber culture system), c-met and m-cadherin were uniformly coexpressed. From the individual satellite cell expression types observed, a model of the satellite cell population at rest and during the time course of activation was generated
P19 Embryonal Carcinoma Cells: A Model System for Studying Neural Tube Induction of Skeletal Myogenesis

AbstractA model experimental system for investigating myogenic induction signals has been devised with mouse P19 embryonal carcinoma cells. When cocultured with pieces of chick neural tube, aggregated P19 cells are induced to become skeletal muscle. The most potent inducing activity is localized to the dorsal neural tube. Less activity was found in the ventral neural tube, notochord, ectoderm, and lateral plate mesoderm, and none was detected in the neural retina. These results suggest that P19 cells may be a useful model system for investigating the mechanisms underlying induction of somite myogenesis
Requirement for Dynamin during Notch Signaling inDrosophilaNeurogenesis

AbstractSingling out of a unique neural precursor from a group of equivalent cells, duringDrosophilaneurogenesis, involves Notch-mediated lateral signaling. During this process, activation of the Notch signaling pathway leads to repression of neural development. Disruption of this signaling pathway results in the development of an excess of neural cells. The loss of activity of dynamin, which is encoded by the geneshibireand is required for endocytosis, results in a similar phenotype. Here we have investigated the requirement ofshibirefunction for Notch signaling during the segregation of sensory bristles on the notum of the fly. Overexpression of different constitutively active forms of Notch inshibiremutant flies indicates thatshibirefunction is not necessary for transduction of the signal downstream of Notch, even when the receptor is integrated in the plasma membrane. However, when wild-type Notch is activated by its ligand Delta, dynamin is required in both signaling and receiving cells for normal singling out of precursors. This suggests an active role of the signaling cell for ligand-mediated receptor endocytosis in the case of transmembrane ligands. We discuss the possible implications of these results for normal functioning of Notch-mediated lateral signaling
Localized Calcium Transients Accompany Furrow Positioning, Propagation, and Deepening during the Early Cleavage Period of Zebrafish Embryos

AbstractThrough the injection off-aequorin (a calcium-specific luminescent reporter) and the use of an imaging photon detector, we see a distinct localized elevation of intracellular calcium that accompanies the appearance of the first furrow arc at the blastodisc surface: the furrow positioning signal. As the leading edges of the arc progress outward toward the margins of the blastodisc, they are accompanied by two subsurface slow calcium waves moving at about 0.2 μm/s: the furrow propagation signal. As these wave fronts approach the edge of the blastodisc, another calcium signal arises in the central region where the positioning signal originally appeared. Like the propagation signal, it extends outward to the margins of the blastodisc, but in this case it also moves downward, accompanying the deepening process that separates the daughter cells: the furrow deepening signal. Both of these furrow deepening progressions move at around 0.1 to 0.2 μm/s. The deepening signal begins to diminish from the center outward, returning to precleavage resting levels on completion of cytokinesis. The signaling sequence is repeated during the second cell division cycle. These localized transients do not require external calcium and they can be dissipated after they have begun by introducing calcium shuttle buffers, resulting in furrow delocalization and regression. They also occur in parthenogenetically activated eggs in which, in an attenuated form, they accompany abortive cleavages
The Dispersion of Clonally Related Cells in the Developing Chick Telencephalon

AbstractLineage analysis in the chick telencephalon was carried out using a library of retroviral vectors. Clones were analyzed in posthatch day 14–21 animals for the phenotype and final locations of sibling cells. Clones often contained multiple types of neurons and glia. Clones of more than four cells almost always crossed functional boundaries. They were dispersed primarily along the rostrocaudal axis or in multiple directions, e.g., along the rostrocaudal and mediolateral axes. In order to begin to understand how the final patterns of dispersion were reached, embryonic tissue was examined. Radial migration, apparently supported by radial glial cells, occurred within the proliferative zones in all clones. In contrast to the migration of cells in the mammalian telencephalon, no tangential migration within the proliferative zones was observed at any age examined. However, beginning at embryonic day 4.5, tangential migration in the mantle zone in multiple directions was observed among the majority of clones. This type of migration occurred as soon as a mantle zone became apparent. It appeared that the tangential migration was not along radial glial processes. As in the mammalian telencephalon and chick diencephalon, dispersion among clonally related cells in the chick telencephalon is frequent, is extensive, and results from tangential migration in a variety of directions