Expression of Sox1 during Xenopus early embryogenesis

Sox B1 group genes, Sox1, Sox2, and Sox3 (Sox1-3), are involved in neurogenesis in various species. Here, we identified the Xenopus homolog of Sox1, and investigated its expression patterns and neural inducing activity. Sox1 was initially expressed in the anterior neural plate of Xenopus embryos, with expression restricted to the brain and optic vesicle by the tailbud stage. Expression subsequently decreased in the eye region by the tadpole stage. Sox1 expression in animal cap explants was induced by inhibition of BMP signaling in the same manner as Sox2, Sox3, and SoxD. In addition, overexpression of Sox1 induced neural markers in ventral ectoderm and in animal caps. These results implicate Xenopus Sox1 in neurogenesis, especially brain and eye development.     

Characterization of <Emphasis Type="Italic">twist</Emphasis> and <Emphasis Type="Italic">snail</Emphasis> gene expression during mesoderm and nervous system development in the polychaete annelid <Emphasis Type="Italic">Capitella</Emphasis> sp. I

To investigate the evolutionary history of mesoderm in the bilaterian lineage, we are studying mesoderm development in the polychaete annelid, Capitella sp. I, a representative lophotrochozoan. In this study, we focus on the Twist and Snail families as candidate mesodermal patterning genes and report the isolation and in situ expression patterns of two twist homologs (CapI-twt1 and CapI-twt2) and two snail homologs (CapI-sna1 and CapI-sna2) in Capitella sp. I. CapI-twt1 is expressed in a subset of mesoderm derivatives during larval development, while CapI-twt2 shows more general mesoderm expression at the same stages. Neither twist gene is detected before the completion of gastrulation. The two snail genes have very distinct expression patterns. At cleavage and early gastrula stages, CapI-sna1 is broadly expressed in precursors of all three germ layers and becomes restricted to cells around the closing blastopore during late gastrulation; CapI-sna2 expression is not detected at these stages. After gastrulation, both snail genes are expressed in the developing central nervous system (CNS) at stages when neural precursor cells are internalized, and CapI-sna1 is also expressed laterally within the segmental mesoderm. Based on the expression patterns in this study, we suggest a putative function for Capitella sp. I twist genes in mesoderm differentiation and for snail genes in regulating CNS development and general cell migration during gastrulation. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Increase in Alphaproteobacteria in association with a polychaete,Capitella sp. I,in the organically enriched sediment

We conducted bioremediation experiments on the organically enriched sediment on the sea floor just below a fish farm, introducing artificially mass-cultured colonies of deposit-feeding polychaete, Capitella sp. I. To clarify the association between the Capitella and bacteria on the efficient decomposition of the organic matter in the sediment in the experiments, we tried to identify the bacteria that increased in the microbial community in the sediment with dense patches of the Capitella. The relationship between TOC and quinone content of the sediment as an indicator of the bacterial abundance was not clear, while a significant positive correlation was found between Capitella biomass and quinone content of the sediment. In particular, ubiquinone-10, which is present in members of the class Alphaproteobacteria, increased in the sediment with dense patches of the Capitella. We performed denaturing gradient gel electrophoresis (DGGE) analyses to identify the alphaproteobacterial species in the sediment with dense patches of the worm, using two DGGE fragments obtained from the sediment samples and one fragment from the worm body. The sequences of these DGGE fragments were closely related to the specific members of the Roseobacter clade. In the associated system with the Capitella and the bacteria in the organically enriched sediment, the decomposition of the organic matter may proceed rapidly. It is very likely that the Capitella works as a promoter of bacteria in the organically enriched sediment, and feeds the increased bacteria as one of the main foods, while the bacteria decompose the organic matter in the sediment with the assistance of the Capitella.     

Assessing Primary Neurogenesis in Xenopus Embryos Using Immunostaining

Primary neurogenesis is a dynamic and complex process during embryonic development that sets up the initial layout of the central nervous system. During this process, a portion of neural stem cells undergo differentiation and give rise to the first populations of differentiated primary neurons within the nascent central nervous system. Several vertebrate model organisms have been used to explore the mechanisms of neural cell fate specification, patterning, and differentiation. Among these is the African clawed frog, Xenopus, which provides a powerful system for investigating the molecular and cellular mechanisms responsible for primary neurogenesis due to its rapid and accessible development and ease of embryological and molecular manipulations. Here, we present a convenient and rapid method to observe the different populations of neuronal cells within Xenopus central nervous system. Using antibody staining and immunofluorescence on sections of Xenopus embryos, we are able to observe the locations of neural stem cells and differentiated primary neurons during primary neurogenesis.     

The ascidian mouth opening is derived from the anterior neuropore: Reassessing the mouth/neural tube relationship in chordate evolution

The relative positions of the brain and mouth are of central importance for models of chordate evolution. The dorsal hollow neural tube and the mouth have often been thought of as developmentally distinct structures that may have followed independent evolutionary paths. In most chordates however, including vertebrates and ascidians, the mouth primordia have been shown to fate to the anterior neural boundary. In ascidians such as Ciona there is a particularly intimate relationship between brain and mouth development, with a thin canal connecting the neural tube lumen to the mouth primordium at larval stages. This so-called neurohypophyseal canal was previously thought to be a secondary connection that formed relatively late, after the independent formation of the mouth primordium and the neural tube. Here we show that the Ciona neurohypophyseal canal is present from the end of neurulation and represents the anteriormost neural tube, and that the future mouth opening is actually derived from the anterior neuropore. The mouth thus forms at the anterior midline transition between neural tube and surface ectoderm. In the vertebrate Xenopus, we find that although the mouth primordium is not topologically continuous with the neural tube lumen, it nonetheless forms at this same transition point. This close association between the mouth primordium and the anterior neural tube in both ascidians and amphibians suggests that the evolution of these two structures may be more closely linked than previously appreciated.     

Effects of sediment-bound Cd, Pb, and Ni on the growth, feeding, and survival of Capitella sp. I

Anthropogenic metal pollutants bioaccumulated in benthic animals by means of feeding and osmotic diffusion. These metals may affect the physiology of the benthos. In this study, we exposed Capitella sp. I to three metals (Cd, Pb, and Ni), each in eight different concentrations, to determine the effects of metals on the animals. Growth rate, ingestion rate, and percent survival were estimated in three separated experiments. The growth and feeding of the worms were sensitive to even the lowest concentrations of each metal added to the sediments. The lowest observable adverse effect levels for Cd, Ni, and Pb were 0.03, 1.59, and 0.41 μmol g^− 1 sediment, respectively. Growth rates in the elevated metal contaminant treatments decreased drastically at slightly contaminated levels, lessened detrimental effects at moderately contaminated levels, and showed incompensable intoxication at heavily contaminated levels. The trends in ingestion rates were similar to those of growth rates. No significant difference in survivorship was found among the different contaminant levels for any of the three heavy metals. Capitella sp. I was most sensitive to Cd, followed by Ni and Pb, which had similar effects. The rapid physiological responses of Capitella sp. I allowed the animals to survive metal exposure. Sediment productivity remained unchanged at different contamination levels of Ni and Pb, but was drastically reduced at 4.75 μmol g^− 1 Cd in the sediment. This further demonstrated Capitella sp. I can adjust their ingestion rates to maintain constant sediment productivities in moderate pollution conditions; however, when threshold concentration was exceeded, homeostasis collapsed.     

Morphological features and viability of Scrippsiella trochoidea cysts isolated from fecal pellets of the polychaete Capitella sp.

To investigate the impact of grazing by the polychaete Capitella sp. on the two cyst morphotypes of Scrippsiella trochoidea, the typical morphotype with short calcareous spines (spiny-type cyst) and artificially induced the transparent type without calcareous spines (naked-type cyst), we examined the morphological features and germination capability of the two cyst morphotypes isolated from fecal pellets of the polychaete Capitella sp. produced in a restricted habitat. The morphological destruction was observed in both spiny- and naked type cysts after passage through the gut of Capitella sp., and this seemed to occur rapidly for naked-type cysts. In addition, the germination of both spiny- and naked-type cysts isolated from fecal pellets on day 2 of harvesting was significantly reduced and subsequently completely abolished, in contrast to previous findings from ingestion studies. Our results indicate that continual grazing by Capitella sp. within a restricted habitat can compromise the survival of S. trochoidea cysts.     

Fine Structure of the Stoma of Bunonema sp. and Teratorhabditis palmarum (Nematoda) and Its Phylogenetic Significance

Fine structure of the stoma, including the cheilostom, gymnostom, and stegostom of Bunonema sp. and Teratorhabditis palmarum was compared with Caenorhabditis elegans to consider fine structural characters that may be phylogenetically informative. The stegostom, enclosed by the anterior end of the pharynx, includes a triradiate lumen surrounded by radial cells (interradial or pairs of adradial cells) repeated in the dorsal and subventral sectors; in Rhabditina, typically the stegostom includes anteriorly two sets of epithelial and posteriorly two sets of muscular radial cells. These muscle cells are anteriorly m1 and posteriorly m2. In Bunonema sp., unlike T. palmarum and C. elegans, the stegostom has a third set of interradial epithelial cells. In Bunonema sp., m1 is expressed by three interradial cells, whereas in T. palmarum and C. elegans m1 is three pairs of adradial muscle cells (i.e., six cells). In all three taxa m2 is expressed as three pairs of adradial muscle cells. Posterior processes of adjacent adradial cells fuse, and closely apposed nuclei may present a figure-eight shape. However, in Bunonema the three interradial m1 cells each have a long posterior process enclosing two separate round nuclei. In combination with additional characters, these diverse stoma features may prove phylogenetically informative. Specifically, the radial epithelial cells of the stegostom appear to be a synapomorphy consistent with a bunonemid-diplogastrid-rhabditid clade, whereas a thickening in the dorsal sector of the stoma cuticle lining is interpreted as a synapomorphy supporting a bunonemid-diplogastrid clade.     

Ordered progression of nematogenesis from stem cells through differentiation stages in the tentacle bulb of Clytia hemisphaerica (Hydrozoa, Cnidaria)

Nematogenesis, the production of stinging cells (nematocytes) in Cnidaria, can be considered as a model neurogenic process. Most molecular data concern the freshwater polyp Hydra, in which nematocyte production is scattered throughout the body column ectoderm, the mature cells then migrating to the tentacles. We have characterized tentacular nematogenesis in the Clytia hemisphaerica hydromedusa and found it to be confined to the ectoderm of the tentacle bulb, a specialized swelling at the tentacle base. Analysis by a variety of light and electron microscope techniques revealed that while cellular aspects of nematogenesis are similar to Hydra, the spatio-temporal characteristics are markedly more ordered. The tentacle bulb nematogenic ectoderm (TBE) was found to be polarized, with a clear progression of successive nematoblast stages from a proximal zone (comprising a majority of undifferentiated cells) to the distal end where the tentacle starts. Pulse-chase labelling experiments demonstrated a continuous displacement of differentiating nematoblasts towards the tentacle tip, and that nematogenesis proceeds more rapidly in Clytia than in Hydra. Compact expression domains of orthologues of known nematogenesis-associated genes (Piwi, dickkopf-3, minicollagens and NOWA) were correspondingly staggered along the TBE. These distinct characteristics make the Clytia TBE a promising experimental system for understanding the mechanisms regulating nematogenesis.     

Mesoderm induction in the future tail region of Xenopus

Summary We have used interspecific grafts between Xenopus borealis and Xenopus laevis to study the signalling system that produces tail mesoderm. Early gastrula ectoderm grafted into the posterior neural plate region of neurulae responds to a mesodermal inducing signal in this region and forms mainly tail somites; this signal persists until at least the early tail bud stage. Ventral ectoderm grafted into the posterior neural plate loses its competence to respond to this signal after stage 10 1/2. We have established the specification of anterior and posterior neural plate ectoderm. In ectodermal sandwiches or when grafted into unusual positions, anterior regions gave rise to mainly nervous system and posterior regions to large amounts of muscle, together with some nervous system. Thus it was impossible to assess the competence of posterior neural plate ectoderm to form further mesoderm and hence to establish if mesodermal induction continues during neurulation in unmanipulated embryos.