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William C Smith

Publications and source records attributed to William C Smith.

15 recordsLinked to original sources
PubMed2006

ADMP2 is essential for primitive blood and heart development in Xenopus.

We describe here the cloning of a new member of the TGF-beta family with similarity to the anti-dorsalizing morphogenetic proteins (ADMPs). This new gene, ADMP2, is expressed in a broad band of mesendoderm cells that appear to include the progenitors of the endoderm and the ventral mesoderm. Antisense morpholino oligonucleotide knockdown of ADMP2 results in near-complete disruption of primitive blood and heart development, while the development of other mesoderm derivatives, including pronephros, muscle and lateral plate is not disrupted. Moreover, the development of the primitive blood in ADMP2 knockdown embryos cannot be rescued by BMP. These results suggests that ADMP2 plays an early role in specifying presumptive ventral mesoderm in the leading edge mesoderm, and that ADMP2 activity may be necessary to respond to BMP signaling in the context of ventral mesoderm induction.

Amino Acid Sequence↗
PubMed2005

Ascidian prickle regulates both mediolateral and anterior-posterior cell polarity of notochord cells.

The ascidian notochord follows a morphogenetic program that includes convergent extension (C/E), followed by anterior-posterior (A/P) elongation [1-4]. As described here, developing notochord cells show polarity first in the mediolateral (M/L) axis during C/E, and subsequently in the A/P axis during elongation. Previous embryological studies [3] have shown that contact with neighboring tissues is essential for directing M/L polarity of ascidian notochord cells. During C/E, the planar cell polarity (PCP) gene products prickle (pk) and dishevelled (dsh) show M/L polarization. pk and dsh colocalize at the notochord cell membranes, with the exception of those in contact with neighboring muscle cells. In the mutant aimless (aim), which carries a deletion in pk, notochord morphogenesis is disrupted, and cell polarization is lost. After C/E, there is a dynamic relocalization of PCP proteins in the notochord cells with dsh localized to the lateral edges of the membrane, and pk and strabismus (stbm) at the anterior edges. An A/P polarity is present in the extending notochord cells and is evident by the position of the nuclei, which in normal embryos are invariably found at the posterior edge of each cell. In the aim mutant, all appearances of A/P polarity in the notochord are lost.

Adaptor Proteins, Signal Transducing↗
PubMed2005

Did the first chordates organize without the organizer?

Models of vertebrate development frequently portray the organizer as acting on a largely unpatterned embryo to induce major components of the body plan, such as the neural plate and somites. Recent experiments examining the molecular and genetic basis of major inductive events of vertebrate embryogenesis force a re-examination of this view. These newer observations, along with a proposed revised fate map for the frog Xenopus laevis, suggest a possible reconciliation between the seemingly disparate mechanisms present in the ontogeny of the common chordate body plan of vertebrate and invertebrate chordates. Here, we review data from vertebrates and from an ascidian urochordate and propose that the organizer was not present at the base of the chordate lineage, but could have been a later innovation in the lineage leading to vertebrates, where its role was more permissive than instructive.

Animals↗
PubMed2005

Pigmentation in the sensory organs of the ascidian larva is essential for normal behavior.

Free-living animals and their larvae utilize light and gravity as cues to navigate in open space. Detection and response to these environmental stimuli are important for the dispersal and settlement of ascidian larvae. Two pigmented structures in the brain of the ascidian larva, the ocellus and the otolith, have been shown to function as the photoreceptive and gravity sensitive organs, respectively. Here, we show that pigmentation is essential for proper phototactic and geotactic behavior in larvae of the ascidian species Ciona savignyi. Two recessive and complementing mutant lines of C. savignyi, immaculate and spotless, that specifically disrupt the pigmentation of the sensory organs during larval development are described. Homozygous mutant larvae are unable to respond properly to gravity and illumination cues while settling. Genetic analysis shows that spotless is caused by a point mutation within the tyrosinase gene that creates a premature stop codon, while the molecular nature of immaculate is unknown. Although the role of pigmentation in the ocellus of C. savignyi is similar to that in vertebrate visual systems, our results demonstrate a novel use of melanin in geotactic behavior.

Animals↗
PubMed2005

Self- and cross-fertilization in the solitary ascidian Ciona savignyi.

Solitary ascidians are hermaphrodites that release sperm and eggs simultaneously. However, many species are self-sterile, owing to a self/non-self recognition system operating at the outer surface of the chorion during sperm-egg interaction. In Ciona intestinalis, self-incompatibility is thought to have a genetic basis. Here, we report a survey of the self-fertilization potential of a Santa Barbara, California, population of Ciona savignyi, a close relative of C. intestinalis. We found that, in contrast to reports on C. intestinalis, C. savignyi is highly self-fertile. However, using two nonlethal recessive mutant strains, aimless (aim) and immaculate (imc), and a stable transgenic strain that expresses green fluorescent protein (GFP) in the notochord to follow offspring genotype, we demonstrate that non-self sperm outcompete self-sperm in fertilization competition assays. When the chorion was removed, both self- and non-self sperm performed equally well in the competition assay. Thus the non-self/self gamete recognition in C. savignyi is not absolute but relative, and is mediated by one or more components in the chorion. We discuss the significance of this finding in the context of natural populations in the wild, where individuals of C. savignyi are typically found growing in large groups that spawn in unison and where self-fertilization would be expected to be very rare.

Animals↗
PubMed2004

Noncoding regulatory sequences of Ciona exhibit strong correspondence between evolutionary constraint and functional importance.

We show that sequence comparisons at different levels of resolution can efficiently guide functional analyses of regulatory regions in the ascidians Ciona savignyi and Ciona intestinalis. Sequence alignments of several tissue-specific genes guided discovery of minimal regulatory regions that are active in whole-embryo reporter assays. Using the Troponin I (TnI) locus as a case study, we show that more refined local sequence analyses can then be used to reveal functional substructure within a regulatory region. A high-resolution saturation mutagenesis in conjunction with comparative sequence analyses defined essential sequence elements within the TnI regulatory region. Finally, we found a significant, quantitative relationship between function and sequence divergence of noncoding functional elements. This work demonstrates the power of comparative sequence analysis between the two Ciona species for guiding gene regulatory experiments.

Animals↗
PubMed2004

A functional cellulose synthase from ascidian epidermis.

Among animals, urochordates (e.g., ascidians) are unique in their ability to biosynthesize cellulose. In ascidians cellulose is synthesized in the epidermis and incorporated into a protective coat know as the tunic. A putative cellulose synthase-like gene was first identified in the genome sequences of the ascidian Ciona intestinalis. We describe here a cellulose synthase gene from the ascidian Ciona savignyi that is expressed in the epidermis. The predicted C. savignyi cellulose synthase amino acid sequence showed conserved features found in all cellulose synthases, including plants, but was most similar to cellulose synthases from bacteria, fungi, and Dictyostelium discoidium. However, unlike other known cellulose synthases, the predicted C. savignyi polypeptide has a degenerate cellulase-like region near the carboxyl-terminal end. An expression construct carrying the C. savignyi cDNA was found to restore cellulose biosynthesis to a cellulose synthase (CelA) minus mutant of Agrobacterium tumefaciens, showing that the predicted protein has cellulose synthase activity. The lack of cellulose biosynthesis in all other groups of metazoans and the similarity of the C. savignyi cellulose synthase to enzymes from cellulose-producing organisms support the hypothesis that the urochordates acquired the cellulose biosynthetic pathway by horizontal transfer.

Agrobacterium tumefaciens↗
PubMed2003

Generation of Ci-Brachyury-GFP stable transgenic lines in the ascidian Ciona savignyi.

We report generation of stable transgenic lines of the ascidian Ciona savignyi carrying a Ciona intestinalis-Brachyury-promoter/Green Fluorescent Protein-reporter (Ci-Bra-GFP) construct. The transgenic lines were made using a technique in which the endonuclease I-SceI was coinjected into fertilized eggs with a transgene construct containing flanking recognition sites for I-SceI. Two founder animals, out of 12 F(0) adults tested, were found to transmit the transgene to their offspring (F(1)s) at frequencies of 42% and 23%. The transgene was further inherited by the F(2) in a Mendelian fashion and displayed nonmosaic expression, indicating integration into the genome. The Mendelian inheritance and the absence of mosaicism persisted through the F(3) and F(4) generations. Southern blot analyses showed that the transgene was organized in tandem arrays of no more than 10 copies. Using these Ci-Bra-GFP transgenics, we describe cellular movements and shape changes involved in notochord morphogenesis in both wildtype and mutant embryos.

Animals↗
PubMed2002

Pitx genes in Tunicates provide new molecular insight into the evolutionary origin of pituitary.

We have initiated a project aimed at documenting molecular and cellular changes underlying the emergence of the hypothalamo-hypophyseal axis in Chordates. Considering the phylogenetic position of Tunicates and the 'pan-hypophyseal' expression pattern of Pitx genes in Vertebrate pituitary, we searched for a Pitx-related homeobox gene in the ascidian Ciona intestinalis, and identified Ci-Pitx (ona intestinalis uitary homeobo gene). We also isolated Cs-Pitx and Bs-Pitx, the Ci-Pitx respective counterparts of Ciona savignyi and Botryllus schlosseri, two other Tunicate species. Ci-Pitx mRNA encodes a putative protein exhibiting the diagnostic K50-Paired-class homeodomain and a conserved C-terminal Aristaless domain. Embryonic expression pattern of Ci-Pitx revealed a conserved expression domain in the anterior neural ridge and subsequently in the pharyngeal primordium, defined in Vertebrates as the stomodeal ectomere, which encompasses the presumptive pituitary territory. This shows that expression at early steps of pituitary development is a feature of Pitx-related genes that was already present in the last common ancestor of Chordates.

Amino Acid Sequence↗
PubMed2002

Expression cloning in ascidians: isolation of a novel member of the asctacin protease family.

The small genome size and gene number of ascidians makes them an ideal model system in which to screen for conserved genes that regulate the development of chordates. Expression cloning has proven to be an effective strategy for isolating genes that play a role in embryogenesis. We have taken advantage of the large size and ease of manipulation of Xenopus embryos for use as an assay system to screen for developmental regulatory genes from the ascidian Ciona intestinalis. Many invertebrate genes have been shown to function in vertebrates, providing us with precedent for our cross-species analysis. The first clone isolated from this screen is an astacin class metalloprotease. This ascidian astacin, named no va, causes a gastrulation defect in Xenopus. In C. intestinalis, no va is expressed both maternally and zygotically. The zygotic expression is seen in the mesenchyme of gastrula and neurula staged embryos.

Amino Acid Sequence↗
PubMed2002

Revisions to the Xenopus gastrula fate map: implications for mesoderm induction and patterning.

A revised fate map of the gastrula Xenopus embryo predicts the existence of patterning mechanisms that operate within the animal/vegetal axis of the mesoderm-forming marginal zone. We review here molecular and embryologic data that demonstrate that such mechanisms are present and that they operate independently of the Spemann organizer. Evidence suggests that polarized fibroblast growth factor activity in the animal/vegetal axis patterns this axis. We present a model of mesoderm induction and patterning that integrates the new data on Spemann organizer-independent animal/vegetal patterning with data on other inductive pathways known to act on the gastrula marginal zone.

Animals↗
PubMed2002

An ascidian engrailed gene.

The engrailed genes play roles in the maintenance of segment polarity in a variety of animals and in the establishment and maintenance of the mid-brain/hind-brain boundary (MHB) in vertebrates. We isolated an ascidian engrailed gene and analyzed its expression pattern during early development. Expression begins at the neurula stage and is restricted to two cells within the neuroectoderm. At the tail-bud stage engrailed-expressing cells are in the "neck" region of the neural tube, which has been proposed to be the ascidian equivalent of the MHB. These same cells also express PAX2/5/8. We speculate that a structure equivalent to the MHB existed before the split of the three chordate sub-phyla.

Amino Acid Sequence↗
PubMed2002

The nodal target gene Xmenf is a component of an FGF-independent pathway of ventral mesoderm induction in Xenopus.

The interplay of fibroblast growth factor (FGF) and nodal signaling in the Xenopus gastrula marginal zone specifies distinct populations of presumptive mesodermal cells. Cells in the vegetal marginal zone, making up the presumptive leading edge mesoderm, are exposed to nodal signaling, as evidenced by SMAD2 activation, but do not appear to be exposed to FGF signaling, as evidenced by the lack of MAP kinase (MAPK) activation. However, in the animal marginal zone, activation of both SMAD2 and MAPK occurs. The differential activation of these two signaling pathways in the marginal zone results in the vegetal and animal marginal zones expressing different genes at gastrulation, and subsequently having different fates, with the vegetal marginal zone contributing to ventral mesoderm (e.g. ventral blood island) and the animal marginal zone giving rise to dorsal fates (e.g. notochord and somite). We report here the cloning of a cDNA encoding a novel nuclear protein, Xmenf, that is expressed in the vegetal marginal zone. The expression of Xmenf is induced by nodal signaling and negatively regulated by FGF signaling. Results from animal cap studies indicate that Xmenf plays a role in the pathway of ventral mesoderm induction in the vegetal marginal zone.

Activins↗