200 results on '"McClay, David R."'
Search Results
2. A Genomic Regulatory Network for Development
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Davidson, Eric H., Rast, Jonathan P., Oliveri, Paola, Ransick, Andrew, Calestani, Cristina, Yuh, Chiou-Hwa, Minokawa, Takuya, Amore, Gabriele, Hinman, Veronica, Arenas-Mena, César, Otim, Ochan, Brown, C. Titus, Livi, Carolina B., Lee, Pei Yun, Revilla, Roger, Rust, Alistair G., Pan, Zheng jun, Schilstra, Maria J., Arnone, Maria I., Rowen, Lee, Cameron, R. Andrew, McClay, David R., Hood, Leroy, and Bolouri, Hamid
- Published
- 2002
3. An RNA interference approach for functional studies in the sea urchin and its use in analysis of nodal signaling gradients
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Wilson, Keen, Manner, Carl, Miranda, Esther, Berrio, Alejandro, Wray, Gregory A., and McClay, David R.
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- 2024
- Full Text
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4. Molecular Cloning of the First Metazoan β -1,3 Glucanase from Eggs of the Sea Urchin Strongylocentrotus purpuratus
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Bachman, Eric Scott and McClay, David R.
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- 1996
5. Spatial and temporal patterns of gene expression during neurogenesis in the sea urchin Lytechinus variegatus
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Slota, Leslie A., Miranda, Esther M., and McClay, David R.
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- 2019
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6. Feedback circuits are numerous in embryonic gene regulatory networks and offer a stabilizing influence on evolution of those networks.
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Massri, Abdull Jesus, McDonald, Brennan, Wray, Gregory A., and McClay, David R.
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CELL differentiation ,STRONGYLOCENTROTUS purpuratus ,REGULATOR genes ,SEA urchins ,LINE drivers (Integrated circuits) ,GENE regulatory networks - Abstract
The developmental gene regulatory networks (dGRNs) of two sea urchin species, Lytechinus variegatus (Lv) and Strongylocentrotus purpuratus (Sp), have remained remarkably similar despite about 50 million years since a common ancestor. Hundreds of parallel experimental perturbations of transcription factors with similar outcomes support this conclusion. A recent scRNA-seq analysis suggested that the earliest expression of several genes within the dGRNs differs between Lv and Sp. Here, we present a careful reanalysis of the dGRNs in these two species, paying close attention to timing of first expression. We find that initial expression of genes critical for cell fate specification occurs during several compressed time periods in both species. Previously unrecognized feedback circuits are inferred from the temporally corrected dGRNs. Although many of these feedbacks differ in location within the respective GRNs, the overall number is similar between species. We identify several prominent differences in timing of first expression for key developmental regulatory genes; comparison with a third species indicates that these heterochronies likely originated in an unbiased manner with respect to embryonic cell lineage and evolutionary branch. Together, these results suggest that interactions can evolve even within highly conserved dGRNs and that feedback circuits may buffer the effects of heterochronies in the expression of key regulatory genes. [ABSTRACT FROM AUTHOR]
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- 2023
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7. Specification to Biomineralization : Following a Single Cell Type as It Constructs a Skeleton
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Lyons, Deirdre C., Martik, Megan L., Saunders, Lindsay R., and McClay, David R.
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- 2014
8. On the Ultrastructure of Hyalin, A Cell Adhesion Protein of the Sea Urchin Embryo Extracellular Matrix
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Adelson, David L., Alliegro, Mark C., and McClay, David R.
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- 1992
9. Intercellular Recognition: Quantitation of Initial Binding Events
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McClay, David R., Wessel, Gary M., and Marchase, Richard B.
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- 1981
10. Echinonectin: A New Embryonic Substrate Adhesion Protein
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Alliegro, Mark C., Ettensohn, Charles A., Burdsal, Carol A., Erickson, Harold P., and McClay, David R.
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- 1988
11. A Requirement for Trypsin-Sensitive Cell-Surface Components for Cell-Cell Interactions of Embryonic Neural Retina Cells
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McClay, David R., Gooding, Linda R., and Fransen, Margaret E.
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- 1977
12. Cell Adhesion to Fibronectin and Tenascin: Quantitative Measurements of Initial Binding and Subsequent Strengthening Response
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Lotz, Margaret M., Burdsal, Carol A., Erickson, Harold P., and McClay, David R.
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- 1989
13. Molecular Heterochronies and Heterotopies in Early Echinoid Development
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Wray, Gregory A. and McClay, David R.
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- 1989
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14. An Autoradiographic Analysis of the Species Specificity during Sponge Cell Reaggregation
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McClay, David R.
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- 1971
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15. Hedgehog Signaling Requires Motile Cilia in the Sea Urchin
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Warner, Jacob F., McCarthy, Ali M., Morris, Robert L., and McClay, David R.
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- 2014
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16. Hedgehog signaling patterns mesoderm in the sea urchin
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Walton, Katherine D., Warner, Jacob, Hertzler, Philip H., and McClay, David R.
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Biological sciences - Abstract
To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.ydbio.2009.04.018 Byline: Katherine D. Walton (a), Jacob Warner (a), Philip H. Hertzler (b), David R. McClay (a) Keywords: Hedgehog; Patched; Smoothened; Morphogenesis; Endoderm; Mesoderm; Signal transduction Abstract: The Hedgehog (Hh) signaling pathway is essential for patterning many structures in vertebrates including the nervous system, chordamesoderm, limb and endodermal organs. In the sea urchin, a basal deuterostome, Hh signaling is shown to participate in organizing the mesoderm. At gastrulation the Hh ligand is expressed by the endoderm downstream of the Brachyury and FoxA transcription factors in the endomesoderm gene regulatory network. The co-receptors Patched (Ptc) and Smoothened (Smo) are expressed by the neighboring skeletogenic and non-skeletogenic mesoderm. Perturbations of Hh, Ptc and Smo cause embryos to develop with skeletal defects and inappropriate non-skeletogenic mesoderm patterning, although initial specification of mesoderm occurs without detectable abnormalities. Perturbations of the pathway caused late defects in skeletogenesis and in the non-skeletogenic mesoderm, including altered numbers of pigment and blastocoelar cells, randomized left-right asymmetry of coelomic pouches, and disorganized circumesophageal muscle causing an inability to swallow. Together the data support the requirement of Hh signaling in patterning each of the mesoderm subtypes in the sea urchin embryo. Author Affiliation: (a) Department of Biology, Duke University, Box 90338, Durham, NC 27708, USA (b) Department of Biology, Central Michigan University, Mount Pleasant, MI 48859, USA Article History: Received 23 May 2008; Revised 30 March 2009; Accepted 17 April 2009
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- 2009
17. Twist is an essential regulator of the skeletogenic gene regulatory network in the sea urchin embryo
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Wu, Shu-Yu, Yang, Yu-Ping, and McClay, David R.
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Genetic research ,Biological sciences - Abstract
To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.ydbio.2008.04.003 Byline: Shu-Yu Wu, Yu-Ping Yang, David R. McClay Keywords: Twist; Mesoderm; Epithelial-mesenchymal transition; Gene regulatory network; Primary mesenchyme cell; Skeletogenesis Abstract: Recent work on the sea urchin endomesoderm gene regulatory network (GRN) offers many opportunities to study the specification and differentiation of each cell type during early development at a mechanistic level. The mesoderm lineages consist of two cell populations, primary and secondary mesenchyme cells (PMCs and SMCs). The micromere-PMC GRN governs the development of the larval skeleton, which is the exclusive fate of PMCs, and SMCs diverge into four lineages, each with its own GRN state. Here we identify a sea urchin ortholog of the Twist transcription factor, and show that it plays an essential role in the PMC GRN and later is involved in SMC formation. Perturbations of Twist either by morpholino knockdown or by overexpression result in defects in progressive phases of PMC development, including specification, ingression/EMT, differentiation and skeletogenesis. Evidence is presented that Twist expression is required for the maintenance of the PMC specification state, and a reciprocal regulation between Alx1 and Twist offers stability for the subsequent processes, such as PMC differentiation and skeletogenesis. These data illustrate the significance of regulatory state maintenance and continuous progression during cell specification, and the dynamics of the sequential events that depend on those earlier regulatory states. Author Affiliation: Department of Biology, French Family Science Center, Duke University, Durham, NC 27708, USA Article History: Received 19 September 2007; Revised 16 January 2008; Accepted 1 April 2008
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- 2008
18. Lineage-specific expansions provide genomic complexity among sea urchin GTPases
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Beane, Wendy S., Voronina, Ekaterina, Wessel, Gary M., and McClay, David R.
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Anopheles -- Physiological aspects ,Anopheles -- Analysis ,Membrane proteins -- Physiological aspects ,Membrane proteins -- Analysis ,Animal genetics -- Physiological aspects ,Animal genetics -- Analysis ,Universities and colleges -- Physiological aspects ,Universities and colleges -- Analysis ,G proteins -- Physiological aspects ,G proteins -- Analysis ,Protein biosynthesis -- Physiological aspects ,Protein biosynthesis -- Analysis ,Biological sciences - Abstract
To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.ydbio.2006.08.046 Byline: Wendy S. Beane (a), Ekaterina Voronina (b), Gary M. Wessel (b), David R. McClay (a) Keywords: Ras GTPase; Rho; G protein; Galpha; Dynamin; SRP; Signal recognition particle receptor; Translation; Gene duplication; Phylogenomics; Echinoderm Abstract: In every organism, GTP-binding proteins control many aspects of cell signaling. Here, we examine in silico several GTPase families from the Strongylocentrotus purpuratus genome: the monomeric Ras superfamily, the heterotrimeric G proteins, the dynamin superfamily, the SRP/SR family, and the 'protein biosynthesis' translational GTPases. Identified were 174 GTPases, of which over 90% are expressed in the embryo as shown by tiling array and expressed sequence tag data. Phylogenomic comparisons restricted to Drosophila, Ciona, and humans (protostomes, urochordates, and vertebrates, respectively) revealed both common and unique elements in the expected composition of these families. G[alpha] and dynamin families contain vertebrate expansions, consistent with whole genome duplications, whereas SRP/SR and translational GTPases are highly conserved. Unexpectedly, Ras superfamily analyses revealed several large (5+) lineage-specific expansions in the sea urchin. For Rho, Rab, Arf, and Ras subfamilies, comparing total human gene numbers to the number of sea urchin genes with vertebrate orthologs suggests reduced genomic complexity in the sea urchin. However, gene duplications in the sea urchin increase overall numbers such that total sea urchin gene numbers approximate vertebrate gene numbers for each monomeric GTPase family. These findings suggest that lineage-specific expansions may be an important component of genomic evolution in signal transduction. Author Affiliation: (a) Department of Biology, Developmental, Cell and Molecular Group, Duke University, Box 91000, Durham, NC 27708, USA (b) Department of Molecular and Cell Biology and Biochemistry, Brown University, 69 Brown Street, Providence, RI 02912, USA Article History: Received 11 May 2006; Revised 18 August 2006; Accepted 19 August 2006
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- 2006
19. Genomics and expression profiles of the Hedgehog and Notch signaling pathways in sea urchin development
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Walton, Katherine D., Croce, Jenifer C., Glenn, Thomas D., Wu, Shu-Yu, and McClay, David R.
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Genetic research ,RNA ,Cell research ,Biological sciences - Abstract
To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.ydbio.2006.08.064 Byline: Katherine D. Walton, Jenifer C. Croce, Thomas D. Glenn, Shu-Yu Wu, David R. McClay Keywords: Hedgehog; Notch; Endoderm; Mesoderm; Sea urchin Abstract: The Hedgehog (Hh) and Notch signal transduction pathways control a variety of developmental processes including cell fate choice, differentiation, proliferation, patterning and boundary formation. Because many components of these pathways are conserved, it was predicted and confirmed that pathway components are largely intact in the sea urchin genome. Spatial and temporal location of these pathways in the embryo, and their function in development offer added insight into their mechanistic contributions. Accordingly, all major components of both pathways were identified and annotated in the sea urchin Strongylocentrotus purpuratus genome and the embryonic expression of key components was explored. Relationships of the pathway components, and modifiers predicted from the annotation of S. purpuratus, were compared against cnidarians, arthropods, urochordates, and vertebrates. These analyses support the prediction that the pathways are highly conserved through metazoan evolution. Further, the location of these two pathways appears to be conserved among deuterostomes, and in the case of Notch at least, display similar capacities in endomesoderm gene regulatory networks. RNA expression profiles by quantitative PCR and RNA in situ hybridization reveal that Hedgehog is produced by the endoderm beginning just prior to invagination, and signals to the secondary mesenchyme-derived tissues at least until the pluteus larva stage. RNA in situ hybridization of Notch pathway members confirms that Notch functions sequentially in the vegetal-most secondary mesenchyme cells and later in the endoderm. Functional analyses in future studies will embed these pathways into the growing knowledge of gene regulatory networks that govern early specification and morphogenesis. Author Affiliation: Developmental, Cellular, and Molecular Biology Group, Duke University, Durham, NC 27710, USA Article History: Received 10 May 2006; Revised 18 August 2006; Accepted 28 August 2006
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- 2006
20. The genomic underpinnings of apoptosis in Strongylocentrotus purpuratus
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Robertson, Anthony J., Croce, Jenifer, Carbonneau, Seth, Voronina, Ekaterina, Miranda, Esther, McClay, David R., and Coffman, James A.
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Sea urchins -- Genetic aspects ,Cell death -- Analysis ,Genomes -- Analysis ,Apoptosis -- Analysis ,Biological sciences - Abstract
A study on the death of cell of sea urchins examines the proteins found in its genome.
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- 2006
21. Lineage-specific expansions provide genomic complexity among sea urchin GTPases
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Wendy S. Beane, Voronina, Ekaterina, Wessel, Gary M., and McClay, David R.
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G proteins -- Research ,Sea urchins -- Research ,Sea urchins -- Observations ,Biological sciences - Abstract
A study on cell signaling examines GTP binding proteins in the genome of sea urchin strongylocentrotus purpuratus.
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- 2006
22. A genome-wide survey of the evolutionarily conserved Wnt pathways in the sea urchin Strongylocentrotus purpuratus
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Croce, Jenifer C., Wu, Shu-Yu, Byrum, Christine, Xu, Ronghui, Duloquin, Louise, Wikramanayake, Athula H., Gache, Christian, and McClay, David R.
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Echinodermata -- Research ,Echinodermata -- Observations ,Genes -- Research ,Sea urchins -- Research ,Cladistic analysis -- Usage ,Biological sciences - Abstract
A study examines the Wnt genes present in the genome of echinoderm Strongylocentrotus purpuratus. A phylogenetic study of the proteins present in the gene of the organism is discussed.
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- 2006
23. RhoA regulates initiation of invagination, but not convergent extension, during sea urchin gastrulation
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Beane, Wendy S., Gross, Jeffrey M., and McClay, David R.
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Muscle proteins -- Analysis ,Biological sciences - Abstract
To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.ydbio.2005.12.031 Byline: Wendy S. Beane (a), Jeffrey M. Gross (b), David R. McClay (a) Keywords: Sea urchin; RhoA; Gastrulation; Invagination; Convergent extension; Brachyury Abstract: During gastrulation, the archenteron is formed using cell shape changes, cell rearrangements, filopodial extensions, and convergent extension movements to elongate and shape the nascent gut tube. How these events are coordinated remains unknown, although much has been learned from careful morphological examinations and molecular perturbations. This study reports that RhoA is necessary to trigger archenteron invagination in the sea urchin embryo. Inhibition of RhoA results in a failure to initiate invagination movements, while constitutively active RhoA induces precocious invagination of the archenteron, complete with the actin rearrangements and extracellular matrix secretions that normally accompany the onset of invagination. Although RhoA activity has been reported to control convergent extension movements in vertebrate embryos, experiments herein show that RhoA activity does not regulate convergent extension movements during sea urchin gastrulation. Instead, the results support the hypothesis that RhoA serves as a trigger to initiate invagination, and once initiation occurs, RhoA activity is no longer involved in subsequent gastrulation movements. Author Affiliation: (a) Department of Biology, Developmental, Cell and Molecular Group, Duke University, PO Box 91000, Durham, NC 27708, USA (b) Molecular Cell and Developmental Biology, University of Texas at Austin, 1 University Station A6700, Austin, TX 78712, USA Article History: Received 6 October 2005; Revised 6 December 2005; Accepted 14 December 2005
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- 2006
24. A Fringe-modified Notch signal affects specification of mesoderm and endoderm in the sea urchin embryo
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Peterson, Robert E. and McClay, David R.
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Endoderm -- Research ,Mesoderm -- Research ,Sea urchin embryo -- Research ,Sea urchin embryo -- Genetic aspects ,Developmental biology -- Research ,Biological sciences - Abstract
Fringe proteins are O-fucose-specific [beta]-1,3 N-acetylglucosanlinyltransferases that glycosylate the extracellular EGF repeats of Notch and enable Notch to be activated by the ligand Delta. In the sea urchin, signaling between Delta and Notch is known to be necessary for specification of secondary mesenchyme cells (SMCs). The Lytechinus variegatus Fringe homologue is expressed in both the signaling and receiving cells during this first Delta-Notch signal. Perturbation of Fringe expression through morpholino antisense oligonucleotide (MO) injection results in fewer SMCs but also causes decreased and delayed archenteron invagination. Partial endoderm specification occurs but expression of some endoderm genes is compromised. The data are consistent with a Fringe-requiting Notch signal as one upstream component of archenteron morphogenesis. Finally, Fringe perturbations result in more severe phenotypes than those previously reported for Notch dominant-negative (Lv[N.sup.neg]) injections or reported here for Notch MO (NMO) injections. Injecting a combination of Lv[N.sup.neg] and NMO results in a more severe phenotype than either treatment alone, and this combination phenocopies the fringe MO embryos. Taken together, the results show that Fringe is necessary both for maternal and zygotic Notch signals, and these Notch signals affect specification of mesoderm and endoderm. Keywords: Sea urchin; Fringe; Notch; Specification; Gastrulation; Morphogenesis; Morpholino antisense oligonucleotide
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- 2005
25. LvGroucho and nuclear [beta]-catenin functionally compete for Tcf binding to influence activation of the endomesoderm gene regulatory network in the sea urchin embryo
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Range, Ryan C., Venuti, Judith M., and McClay, David R.
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Genetics -- Research ,Sea urchins -- Research ,Genetic research ,Biological sciences - Abstract
In the sea urchin embryo, specification of the endomesoderm is accomplished by the activity of a network of regulatory genes in the vegetal hemisphere, called the endomesoderm gene regulatory network (GRN). The activation of this network is mediated primarily through the activity of the Wnt pathway, though details of pathway activation remain unclear. To gain further insight into control of endomesoderm GRN activation, we have identified a sea urchin homologue of the co-repressor Groucho (LvGroucho) that has been shown to antagonize [beta]catenin/Tcf activation complexes during Wnt signaling in other systems. Groucho functions by recruiting the histone deacetylase Rpd3 to the DNA template via interaction with site-specific transcription factors, resulting in localized chromatin condensation and transcriptional silencing. Our results show that the LvGroucho protein localizes to all nuclei throughout embryonic development. Interaction assays demonstrate that LvGroucho interacts with Tcf via both the Q and the WD domains of the protein. LvGroucho interacts with Tcf to antagonize the expression of key endomesoderm regulatory genes. Assays demonstrate that LvGroucho and n[beta]-catenin functionally compete for binding to Tcf as a major mechanism by which the Tcf-control switch is regulated. Functional analysis of the N-terminal AES 197 domain of LvGroucho shows that it is sufficient to recapitulate the function of full-length LvGroucho. This finding strongly supports the conclusion that the effects of LvGro overexpression are due primarily to its interactions with Tcf and not other Groucho interacting partners, since Tcf is the only protein present in the sea urchin known to interact with AES197. Because the Q domain is unable to bind Rpd3, it was expected to behave as a dominant negative LvGroucho. Unexpectedly, overexpression of the Q domain gave functional results similar to LvGroucho and the AES197 domain. This is the first evidence for an inherent repressive function for the Q domain alone. Together, our results indicate that LvGroucho functionally competes with [beta]-catenin for Tcf binding, and this competitive mechanism regulates one of the earliest steps in the initiation of the sea urchin endomesoderm GRN. Keywords: Groucho; [beta]-catenin; Tcf; Endoderm; Mesoderm; Cell fate specification; Sea urchin
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- 2005
26. SpHnf6, a transcription factor that executes multiple functions in sea urchin embryogenesis
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Otim, Ochan, Amore, Gabriele, Minokawa, Takuya, McClay, David R., and Davidson, Eric H.
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Developmental biology -- Research ,Sea urchin embryo -- Research ,Sea urchin embryo -- Genetic aspects ,Genetic transcription -- Research ,Biological sciences - Abstract
The Strongylocentrotus purpuratus hnf6 (Sphnf6) gene encodes a new member of the ONECUT family of transcription factors. The expression of hnf6 in the developing embryo is triphasic, and loss-of-function analysis shows that the Hnf6 protein is a transcription factor that has multiple distinct roles in sea urchin development, hnf6 is expressed maternally, and before gastrulation its transcripts are distributed globally. Early in development, its expression is required for the activation of PMC differentiation genes such as sm50, pm27, and msp130, but not for the activation of any known PMC regulatory genes, for example, alx, ets1, pmar1, or tbrain. Micromere transplantation experiments show that the gene is not involved in early micromere signaling. Early hnf6 expression is also required for expression of the mesodermal regulator gatac. The second known role of hnf6 is its participation after gastrulation in the oral ectoderm gene regulatory network (GRN), in which its expression is essential for the maintenance of the state of oral ectoderm specification. The third role is in the neurogenic ciliated band, which is foreshadowed exactly by a trapezoidal band of hnf6 expression at the border of the oral ectoderm and where it continues to be expressed through the end of embryogenesis. Neither oral ectoderm regulatory functions nor ciliated band formation occur normally in the absence of hnf6 expression. Keywords: Morpholino antisense oligonucleotide; Transcription factor; hnf6; ONECUT; Oral ectoderm; PMC; Sea urchin embryo
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- 2004
27. Spdeadringer, a sea urchin embryo gene required separately in skeletogenic and oral ectoderm gene regulatory networks
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Amore, Gabriele, Yavrouian, Robert G., Peterson, Kevin J., Ransick, Andrew, McClay, David R., and Davidson, Eric H.
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Ectoderm -- Research ,Genetic regulation -- Research ,Genetic transcription -- Research ,Sea urchins -- Genetic aspects ,Sea urchins -- Research ,Biological sciences - Abstract
The Spdeadringer (Spdri) gene encodes an ARID-class transcription factor not previously known in sea urchin embryos. We show that Spdri is a key player in two separate developmental gene regulatory networks (GRNs). Spdri is expressed in a biphasic manner, first, after 12 h and until ingression in the skeletogenic descendants of the large micromeres; second, after about 20 h in the oral ectoderm, where its transcripts remain present at 30-50 mRNA molecules/cell far into development. In both territories, the periods of Spdri expression follow prior territorial specification events. The functional significance of each phase of expression was assessed by determining the effect of an [alpha]Spdri morpholino antisense oligonucleotide (MASO) on expression of 17 different mesodermal genes, 8 different oral ectoderm genes, and 18 other genes expressed specifically during endomesoderm specification. These effects were measured by quantitative PCR, supplemented by whole-mount in situ hybridization and morphological observations. Spdri is shown to act in the micromere descendants in the pathways that result in the expression of batteries of terminal skeletogenic genes. But, in the oral ectoderm, the same gene participates in the central GRN controlling oral ectoderm identity. Spdri is linked in the oral ectoderm GRN with several other genes encoding transcriptional regulators that are expressed specifically in various regions of the oral ectoderm. If its expression is blocked by treatment with [alpha]Spdri MASO, oral-specific features disappear and expression of the aboral ectoderm marker spec1 encompasses the whole of the ectoderm. In addition to disappearance of the oral ectoderm, morphological consequences of [alpha]Spdri MASO treatment include failure of spiculogenesis and of correct primary mesenchyme cell (pmc) patterning in the postgastrular embryo, and also failure of gastrulation. To further analyze these phenotypes, chimeric embryos were constructed consisting of two labeled micromeres combined with micromereless 4th cleavage host embryos; either the micromeres or the hosts contained [alpha]Spdri MASO. These experiments showed that, while Spdri expression is required autonomously for expression of skeletogenic genes prior to ingression, complete skeletogenesis also requires the expression of oral ectoderm patterning information. Presentation of this information on the oral side of the blastocoel in turn depends on Spdri expression in the oral ectoderm. Failure of gastrulation is not due to indirect interference with endomesodermal specification per se, since all endomesodermal genes tested function normally in [alpha]Spdri MASO embryos. Part of its cause is interference by [alpha]Spdri MASO with a late signaling function on the part of the micromere descendants that is needed to complete clearance of the Soxb1 repressor of gastrulation from the prospective endoderm, but in addition there is a nonautonomous oral ectoderm effect. Keywords: Gene regulatory network; Oral ectoderm; deadringer; Sea urchin embryo
- Published
- 2003
28. Activation of pmar1 controls specification of micromeres in the sea urchin embryo
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Oliveri, Paola, Davidson, Eric H., and McClay, David R.
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Genetic regulation -- Research ,Genetic transcription -- Research ,Sea urchins -- Genetic aspects ,Sea urchins -- Research ,Biological sciences - Abstract
pmarl is a transcription factor in the paired class homeodomain family that was identified and found to be transcribed in micromeres beginning at the fourth cleavage of sea urchin development [Dev. Biol. 246 (2002), 209]. Based on in situ data, molecular perturbation studies, and QPCR data, the recently published gene regulatory network (GRN) model for endomesoderm specification [Science 295 (2002) 1669; Dev. Biol. 246 (2002), 162] places pmarl early in the micromere specification pathway, and upstream of two important micromere induction signals. The goal of this study was to test these three predictions of the network model. A series of embryo chimeras were produced in which pmarl activity was perturbed in one cell that was transplanted to control hosts. At the fourth cleavage, micromeres bearing altered pmarl activity were combined with a normal micromereless host embryo. If [beta]-catenin signaling is blocked, the micromeres remain unspecified and are unable to signal to the host cells. When such [beta]-catenin-blocked micromeres also express Pmarl, all observed micromere functions are rescued. The rescue includes expression of the primary mesenchyme cell (PMC) differentiation program, expression and execution of the Delta signal to induce secondary mesoderm cell (SMC) specification in macromere progeny, and expression of the early endomesoderm induction signal necessary for full specification of the endoderm. Additionally, Pmarl expressed mosaically from inserted DNA constructs causes induction of ectopic Endo 16 in adjacent cells, demonstrating further that Pmarl controls expression of the early endomesoderm induction signal. Based on these experiments, Pmarl is an important transcription factor necessary for initiating the micromere specification program and for the expression of two inductive signals produced by micromeres. Each of the tests we describe supports the placement and function of Pmarl in the endomesoderm GRN model. Keywords: Gene regulatory network; Sox; Delta; [beta]-Catenin; Paired homeodomain
- Published
- 2003
29. Primary mesenchyme cell patterning during the early stages following ingression
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Peterson, Robert E. and McClay, David R.
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Messenger RNA ,Stem cells ,Biological sciences - Abstract
To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/S0012-1606(02)00025-8 Byline: Robert E Peterson (a), David R McClay (a) Keywords: Ingression; Epithelial to mesenchymal transition; EMT; Sea urchin; Micromere; Primary mesenchyme cells; PMC; Morphogenesis; GFP; mRNA microinjection Abstract: Sea urchin primary mesenchyme cells (PMCs) ingress into the blastocoel during an epithelial-to-mesenchymal transition (EMT), migrate along the blastocoelar wall for a period of time, and then settle into a subequatorial ring to form the larval skeleton. Fluorescent-marked blastomeres alone, or in combination with blastomere recombination, were used to track the position of PMCs during the early phases of this movement. Micromeres expressing Golgi-tethered GFP (galtase-GFP) were transplanted onto TRITC-stained hosts (in place of the endogenous micromere) to observe the progeny of a single micromere. Galtase-GFP as a Golgi marker is not transferred between PMCs when the syncytium forms. Thus, the position of cells can be followed relative to beginning position for longer periods than previously reported. The PMC progeny of a single micromere do not disperse upon ingression, but instead remain in a closely associated cluster. Generally, progeny of a single micromere remain in the quadrant of origin. In total, greater than [approximately equal to]94% of labeled PMCs remain within the local region of ingression. By contrast, when a transplanted micromere is placed at the vegetal plate after removing all 4 host micromeres, the resultant PMCs ingress and migrate into all 4 quadrants. Similarly, if 1 blastomere is injected at the 2-cell stage, and later the 2 unlabeled micromeres are removed at the 16-cell stage, the remaining PMCs ingress into all 4 quadrants of the vegetal plate. We conclude that the normal restriction of PMCs to a quadrant is due to mechanical constraint from other micromere-PMCs. If a labeled micromere is placed ectopically at the macromere/mesomere boundary, the PMC progeny ingress ectopically and migrate longitudinally along the animal-vegetal axis only. Injection of galtase-GFP into one blastomere at the 4-cell stage shows a 2-step pattern of localization. At late mesenchyme blastula and early gastrula stages, greater than 90% of GFP-expressing PMCs remain in the injected quadrant, while at mid- to late-gastrula stage and beyond, more PMCs are found outside the injected quadrant. The migration that sets up the asymmetry of the larval skeleton first occurs around mid- to late-gastrula stages, when some PMCs from an aboral quadrant migrate to the adjacent oral quadrant. In all, these data combined with previous data suggest that freshly ingressed PMCs migrate along a longitudinal path toward the animal pole and back toward the vegetal pole. Beginning at mid- to late-gastrula stage, PMCs utilize oral-aboral cues from the ectoderm for the first time. At this time, some aboral PMCs migrate into the adjacent oral quadrant to assist in the formation of the ventrolateral cluster. Author Affiliation: (a) Duke University, DCMB Group, Durham, NC 27710, USA Article History: Received 3 December 2001; Revised 16 October 2002; Accepted 23 October 2002
- Published
- 2003
30. Developmental single-cell transcriptomics in the Lytechinus variegatus sea urchin embryo.
- Author
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Massri, Abdull J., Greenstreet, Laura, Afanassiev, Anton, Berrio, Alejandro, Wray, Gregory A., Schiebinger, Geoffrey, and McClay, David R.
- Subjects
SEA urchins ,GENE regulatory networks ,EMBRYOS ,ENDODERM ,GASTRULATION - Abstract
Using scRNA-seq coupled with computational approaches, we studied transcriptional changes in cell states of sea urchin embryos during development to the larval stage. Eighteen closely spaced time points were taken during the first 24 h of development of Lytechinus variegatus (Lv). Developmental trajectories were constructed using Waddington-OT, a computational approach to 'stitch' together developmental time points. Skeletogenic and primordial germ cell trajectories diverged early in cleavage. Ectodermal progenitors were distinct from other lineages by the 6th cleavage, although a small percentage of ectoderm cells briefly co-expressed endoderm markers that indicated an early ecto-endoderm cell state, likely in cells originating from the equatorial region of the egg. Endomesoderm cells also originated at the 6th cleavage and this state persisted for more than two cleavages, then diverged into distinct endoderm and mesoderm fates asynchronously, with some cells retaining an intermediate specification status until gastrulation. Seventy-nine out of 80 genes (99%) examined, and included in published developmental gene regulatory networks (dGRNs), are present in the Lv-scRNA-seq dataset and are expressed in the correct lineages in which the dGRN circuits operate. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
31. Regulated exocytosis and sequential construction of the extracellular matrix surrounding the sea urchin zygote
- Author
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Matese, John C., Black, Steven, and McClay, David R.
- Subjects
Antigens ,Biological sciences - Abstract
Byline: John C. Matese, Steven Black, David R. McClay Abstract: After fertilization most eggs become surrounded by a complex extracellular matrix. This study examines those matrix assembly processes that are triggered by fertilization of the sea urchin egg. The study uses antibodies that identify five different storage compartments in the egg. These compartments release their protein contents in a highly regulated fashion to assemble and modify the extraembryonic layers. The exocytosis sequence begins with a fertilization wave that progresses from the site of sperm entry and elevates the fertilization envelope above a water-filled perivitelline space. The immediate surface of the zygote then becomes covered by a newly secreted hyaline layer. Prior to fertilization some of the antigens are localized to cortical granules. Others are found in 'basal laminar vesicles' that are released in a wave beginning at about 30 sec, or roughly at the same time as cortical granule exocytosis. The remaining antigens are exocytosed with a rather precise timing, but with a delay of several to tens of minutes relative to the first wave of exocytosis. 'Apical vesicles,' so named because antigens from this class are preferentially exocytosed toward the apical cell surface of polarized cells, include antigens that are exocytosed beginning at about 5 min postfertilization. The fourth compartment, named 'echinonectin vesicles' release echinonectin, a protein that is deposited to the inner side of the hyaline layer. Surface staining of echinonectin is first detected about 10-15 min following sperm contact. Finally, maternal cadherin, which is stored in yet a fifth distinct compartment, is not detected on the surface until at least 30 min following fertilization. The data are also consistent with the notion that the tightly regulated timing of exocytosis contributes to the ordered assembly of the hyaline layer and elevation of the fertilization envelope. Finally, two of the vesicle classes continue to exocytose after the cells become polarized. In polarized cells apical and basal laminar antigens are trafficked toward opposite sides of the same cell after passing through the same trans-Golgi network-like compartment. Author Affiliation: Developmental, Cell and Molecular Biology Group, Box 91000, Duke University, Durham, North Carolina 27708, USA Article History: Received 24 December 1996; Accepted 7 April 1997
- Published
- 1997
32. A provisional regulatory gene network for specification of endomesoderm in the sea urchin embryo
- Author
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Davidson, Eric H, Rast, Jonathan P, Oliveri, Paola, Ransick, Andrew, Calestani, Cristina, Yuh, Chiou-Hwa, Minokawa, Takuya, Amore, Gabriele, Hinman, Veronica, Arenas-Mena, Cesar, Otim, Ochan, Brown, C. Titus, Livi, Carolina B, Lee, Pei Yun, Revilla, Roger, Schilstra, Maria J, Clarke, Peter J C, Rust, Alistair G, Pan, Zhengjun, Arnone, Maria I, Rowen, Lee, Cameron, R. Andrew, McClay, David R, Hood, Leroy, and Bolouri, Hamid
- Subjects
Life Sciences (General) - Abstract
We present the current form of a provisional DNA sequence-based regulatory gene network that explains in outline how endomesodermal specification in the sea urchin embryo is controlled. The model of the network is in a continuous process of revision and growth as new genes are added and new experimental results become available; see http://www.its.caltech.edu/~mirsky/endomeso.htm (End-mes Gene Network Update) for the latest version. The network contains over 40 genes at present, many newly uncovered in the course of this work, and most encoding DNA-binding transcriptional regulatory factors. The architecture of the network was approached initially by construction of a logic model that integrated the extensive experimental evidence now available on endomesoderm specification. The internal linkages between genes in the network have been determined functionally, by measurement of the effects of regulatory perturbations on the expression of all relevant genes in the network. Five kinds of perturbation have been applied: (1) use of morpholino antisense oligonucleotides targeted to many of the key regulatory genes in the network; (2) transformation of other regulatory factors into dominant repressors by construction of Engrailed repressor domain fusions; (3) ectopic expression of given regulatory factors, from genetic expression constructs and from injected mRNAs; (4) blockade of the beta-catenin/Tcf pathway by introduction of mRNA encoding the intracellular domain of cadherin; and (5) blockade of the Notch signaling pathway by introduction of mRNA encoding the extracellular domain of the Notch receptor. The network model predicts the cis-regulatory inputs that link each gene into the network. Therefore, its architecture is testable by cis-regulatory analysis. Strongylocentrotus purpuratus and Lytechinus variegatus genomic BAC recombinants that include a large number of the genes in the network have been sequenced and annotated. Tests of the cis-regulatory predictions of the model are greatly facilitated by interspecific computational sequence comparison, which affords a rapid identification of likely cis-regulatory elements in advance of experimental analysis. The network specifies genomically encoded regulatory processes between early cleavage and gastrula stages. These control the specification of the micromere lineage and of the initial veg(2) endomesodermal domain; the blastula-stage separation of the central veg(2) mesodermal domain (i.e., the secondary mesenchyme progenitor field) from the peripheral veg(2) endodermal domain; the stabilization of specification state within these domains; and activation of some downstream differentiation genes. Each of the temporal-spatial phases of specification is represented in a subelement of the network model, that treats regulatory events within the relevant embryonic nuclei at particular stages. (c) 2002 Elsevier Science (USA).
- Published
- 2002
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33. Chromosomal-Level Genome Assembly of the Sea Urchin Lytechinus variegatus Substantially Improves Functional Genomic Analyses.
- Author
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Davidson, Phillip L, Guo, Haobing, Wang, Lingyu, Berrio, Alejandro, Zhang, He, Chang, Yue, Soborowski, Andrew L, McClay, David R, Fan, Guangyi, and Wray, Gregory A
- Subjects
SEA urchins ,FUNCTIONAL analysis ,GENE families ,GENOMES ,FUNCTIONAL genomics ,CHROMOSOMES - Abstract
Lytechinus variegatus is a camarodont sea urchin found widely throughout the western Atlantic Ocean in a variety of shallow-water marine habitats. Its distribution, abundance, and amenability to developmental perturbation make it a popular model for ecologists and developmental biologists. Here, we present a chromosomal-level genome assembly of L. variegatus generated from a combination of PacBio long reads, 10× Genomics sequencing, and HiC chromatin interaction sequencing. We show L. variegatus has 19 chromosomes with an assembly size of 870.4 Mb. The contiguity and completeness of this assembly are reflected by a scaffold length N50 of 45.5 Mb and BUSCO completeness score of 95.5%. Ab initio and transcript-informed gene modeling and annotation identified 27,232 genes with an average gene length of 12.6 kb, comprising an estimated 39.5% of the genome. Repetitive regions, on the other hand, make up 45.4% of the genome. Physical mapping of well-studied developmental genes onto each chromosome reveals nonrandom spatial distribution of distinct genes and gene families, which provides insight into how certain gene families may have evolved and are transcriptionally regulated in this species. Lastly, aligning RNA-seq and ATAC-seq data onto this assembly demonstrates the value of highly contiguous, complete genome assemblies for functional genomics analyses that is unattainable with fragmented, incomplete assemblies. This genome will be of great value to the scientific community as a resource for genome evolution, developmental, and ecological studies of this species and the Echinodermata. [ABSTRACT FROM AUTHOR]
- Published
- 2020
- Full Text
- View/download PDF
34. Branching out: origins of the sea urchin larval skeleton in development and evolution
- Author
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McIntyre, Daniel C., Lyons, Deirdre C., Martik, Megan, and McClay, David R.
- Subjects
Vascular Endothelial Growth Factor A ,animal structures ,Biological Evolution ,Models, Biological ,Article ,Calcium Carbonate ,Fibroblast Growth Factors ,Wnt Proteins ,Cell Movement ,Larva ,Sea Urchins ,Ectoderm ,Animals ,Body Patterning ,Signal Transduction - Abstract
It is a challenge to understand how the information encoded in DNA is used to build a three-dimensional structure. To explore how this works the assembly of a relatively simple skeleton has been examined at multiple control levels. The skeleton of the sea urchin embryo consists of a number of calcite rods produced by 64 skeletogenic cells. The ectoderm supplies spatial cues for patterning, essentially telling the skeletogenic cells where to position themselves and providing the factors for skeletal growth. Here, we describe the information known about how this works. First the ectoderm must be patterned so that the signaling cues are released from precise positions. The skeletogenic cells respond by initiating skeletogenesis immediately beneath two regions (one on the right and the other on the left side). Growth of the skeletal rods requires additional signaling from defined ectodermal locations, and the skeletogenic cells respond to produce a membrane-bound template in which the calcite crystal grows. Important in this process are three signals, fibroblast growth factor, vascular endothelial growth factor, and Wnt5. Each is necessary for explicit tasks in skeleton production.
- Published
- 2014
35. Identification of neural transcription factors required for the differentiation of three neuronal subtypes in the sea urchin embryo.
- Author
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Slota, Leslie A. and McClay, David R.
- Subjects
- *
TRANSCRIPTION factors , *NERVOUS system , *SEA urchin embryos , *NOTCH signaling pathway , *SEROTONINERGIC mechanisms - Abstract
Correct patterning of the nervous system is essential for an organism's survival and complex behavior. Embryologists have used the sea urchin as a model for decades, but our understanding of sea urchin nervous system patterning is incomplete. Previous histochemical studies identified multiple neurotransmitters in the pluteus larvae of several sea urchin species. However, little is known about how, where and when neural subtypes are differentially specified during development. Here, we examine the molecular mechanisms of neuronal subtype specification in 3 distinct neural subtypes in the Lytechinus variegatus larva. We show that these subtypes are specified through Delta/Notch signaling and identify a different transcription factor required for the development of each neural subtype. Our results show achaete-scute and neurogenin are proneural for the serotonergic neurons of the apical organ and cholinergic neurons of the ciliary band, respectively. We also show that orthopedia is not proneural but is necessary for the differentiation of the cholinergic/catecholaminergic postoral neurons. Interestingly, these transcription factors are used similarly during vertebrate neurogenesis. We believe this study is a starting point for building a neural gene regulatory network in the sea urchin and for finding conserved deuterostome neurogenic mechanisms. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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- View/download PDF
36. New insights from a high-resolution look at gastrulation in the sea urchin, Lytechinus variegatus.
- Author
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Martik, Megan L. and McClay, David R.
- Subjects
- *
SEA urchins , *GASTRULATION , *CELL division , *CELL migration , *CELL transplantation , *FISHES - Abstract
Background Gastrulation is a complex orchestration of movements by cells that are specified early in development. Until now, classical convergent extension was considered to be the main contributor to sea urchin archenteron extension, and the relative contributions of cell divisions were unknown. Active migration of cells along the axis of extension was also not considered as a major factor in invagination. Results Cell transplantations plus live imaging were used to examine endoderm cell morphogenesis during gastrulation at high-resolution in the optically clear sea urchin embryo. The invagination sequence was imaged throughout gastrulation. One of the eight macromeres was replaced by a fluorescently labeled macromere at the 32 cell stage. At gastrulation those patches of fluorescent endoderm cell progeny initially about 4 cells wide, released a column of cells about 2 cells wide early in gastrulation and then often this column narrowed to one cell wide by the end of archenteron lengthening. The primary movement of the column of cells was in the direction of elongation of the archenteron with the narrowing (convergence) occurring as one of the two cells moved ahead of its neighbor. As the column narrowed, the labeled endoderm cells generally remained as a contiguous population of cells, rarely separated by intrusion of a lateral unlabeled cell. This longitudinal cell migration mechanism was assessed quantitatively and accounted for almost 90% of the elongation process. Much of the extension was the contribution of Veg2 endoderm with a minor contribution late in gastrulation by Veg1 endoderm cells. We also analyzed the contribution of cell divisions to elongation. Endoderm cells in Lytechinus variagatus were determined to go through approximately one cell doubling during gastrulation. That doubling occurs without a net increase in cell mass, but the question remained as to whether oriented divisions might contribute to archenteron elongation. We learned that indeed there was a biased orientation of cell divisions along the plane of archenteron elongation, but when the impact of that bias was analyzed quantitatively, it contributed a maximum 15% to the total elongation of the gut. Conclusions The major driver of archenteron elongation in the sea urchin, Lytechinus variagatus, is directed movement of Veg2 endoderm cells as a narrowing column along the plane of elongation. The narrowing occurs as cells in the column converge as they migrate, so that the combination of migration and the angular convergence provide the major component of the lengthening. A minor contributor to elongation is oriented cell divisions that contribute to the lengthening but no more than about 15%. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
- View/download PDF
37. Morphogenesis in sea urchin embryos: linking cellular events to gene regulatory network states
- Author
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Lyons, Deidre, Kaltenbach, Stacy, and McClay, David R.
- Subjects
animal structures ,Osteogenesis ,Sea Urchins ,Stem Cells ,embryonic structures ,Animals ,Cell Lineage ,Gene Regulatory Networks ,Article - Abstract
Gastrulation in the sea urchin begins with ingression of the primary mesenchyme cells (PMCs) at the vegetal pole of the embryo. After entering the blastocoel the PMCs migrate, form a syncitium, and synthesize the skeleton of the embryo. Several hours after the PMCs ingress the vegetal plate buckles to initiate invagination of the archenteron. That morphogenetic process occurs in several steps. The nonskeletogenic cells produce the initial inbending of the vegetal plate. Endoderm cells then rearrange and extend the length of the gut across the blastocoel to a target near the animal pole. Finally, cells that will form part of the midgut and hindgut are added to complete gastrulation. Later, the stomodeum invaginates from the oral ectoderm and fuses with the foregut to complete the archenteron. In advance of, and during these morphogenetic events, an increasingly complex input of transcription factors controls the specification and the cell biological events that conduct the gastrulation movements.
- Published
- 2011
38. Comparative Developmental Transcriptomics Reveals Rewiring of a Highly Conserved Gene Regulatory Network during a Major Life History Switch in the Sea Urchin Genus Heliocidaris.
- Author
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Israel, Jennifer W., Martik, Megan L., Byrne, Maria, Raff, Elizabeth C., Raff, Rudolf A., McClay, David R., and Wray, Gregory A.
- Subjects
SEA urchin development ,FOSSIL sea urchins ,RNA sequencing ,GENE expression ,GENE regulatory networks ,HISTORY - Abstract
The ecologically significant shift in developmental strategy from planktotrophic (feeding) to lecithotrophic (nonfeeding) development in the sea urchin genus Heliocidaris is one of the most comprehensively studied life history transitions in any animal. Although the evolution of lecithotrophy involved substantial changes to larval development and morphology, it is not known to what extent changes in gene expression underlie the developmental differences between species, nor do we understand how these changes evolved within the context of the well-defined gene regulatory network (GRN) underlying sea urchin development. To address these questions, we used RNA-seq to measure expression dynamics across development in three species: the lecithotroph Heliocidaris erythrogramma, the closely related planktotroph H. tuberculata, and an outgroup planktotroph Lytechinus variegatus. Using well-established statistical methods, we developed a novel framework for identifying, quantifying, and polarizing evolutionary changes in gene expression profiles across the transcriptome and within the GRN. We found that major changes in gene expression profiles were more numerous during the evolution of lecithotrophy than during the persistence of planktotrophy, and that genes with derived expression profiles in the lecithotroph displayed specific characteristics as a group that are consistent with the dramatically altered developmental program in this species. Compared to the transcriptome, changes in gene expression profiles within the GRN were even more pronounced in the lecithotroph. We found evidence for conservation and likely divergence of particular GRN regulatory interactions in the lecithotroph, as well as significant changes in the expression of genes with known roles in larval skeletogenesis. We further use coexpression analysis to identify genes of unknown function that may contribute to both conserved and derived developmental traits between species. Collectively, our results indicate that distinct evolutionary processes operate on gene expression during periods of life history conservation and periods of life history divergence, and that this contrast is even more pronounced within the GRN than across the transcriptome as a whole. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
39. Perturbations to the Hedgehog Pathway in Sea Urchin Embryos.
- Author
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Warner, Jacob F. and McClay, David R.
- Published
- 2014
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40. Evolution of the Wnt Pathways.
- Author
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Croce, Jenifer C. and McClay, David R.
- Published
- 2009
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- View/download PDF
41. Left-right asymmetry in the sea urchin.
- Author
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Warner, Jacob F. and McClay, David R.
- Published
- 2014
- Full Text
- View/download PDF
42. Sub-circuits of a gene regulatory network control a developmental epithelial-mesenchymal transition.
- Author
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Saunders, Lindsay R. and McClay, David R.
- Subjects
- *
GENE regulatory networks , *NUCLEOTIDE sequence , *EPITHELIAL cells , *MESENCHYMAL stem cells , *TRANSCRIPTION factors , *EMBRYOLOGY , *METASTASIS - Abstract
Epithelial-mesenchymal transition (EMT) is a fundamental cell state change that transforms epithelial to mesenchymal cells during embryonic development, adult tissue repair and cancer metastasis. EMT includes a complex series of intermediate cell state changes including remodeling of the basement membrane, apical constriction, epithelial de-adhesion, directed motility, loss of apical-basal polarity, and acquisition of mesenchymal adhesion and polarity. Transcriptional regulatory state changes must ultimately coordinate the timing and execution of these cell biological processes. A wellcharacterized gene regulatory network (GRN) in the sea urchin embryo was used to identify the transcription factors that control five distinct cell changes during EMT. Single transcription factors were perturbed and the consequences followed with in vivo time-lapse imaging or immunostaining assays. The data show that five different sub-circuits of the GRN control five distinct cell biological activities, each part of the complex EMT process. Thirteen transcription factors (TFs) expressed specifically in pre-EMT cells were required for EMT. Three TFs highest in the GRN specified and activated EMT (alx1, ets1, tbr) and the 10 TFs downstream of those (tel, erg, hex, tgif, snail, twist, foxn2/3, dri, foxb, foxo) were also required for EMT. No single TF functioned in all five sub-circuits, indicating that there is no EMT master regulator. Instead, the resulting sub-circuit topologies suggest EMT requires multiple simultaneous regulatory mechanisms: forward cascades, parallel inputs and positive-feedback lock downs. The interconnected and overlapping nature of the sub-circuits provides one explanation for the seamless orchestration by the embryo of cell state changes leading to successful EMT. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
- View/download PDF
43. Short-range Wnt5 signaling initiates specification of sea urchin posterior ectoderm.
- Author
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McIntyre, Daniel C., Seay, N. Winn, Croce, Jenifer C., and McClay, David R.
- Subjects
SEA urchins ,CONNECTIVE tissues ,TRANSCRIPTION factors ,HEREDITY ,CELLS - Abstract
The border between the posterior ectoderm and the endoderm is a location where two germ layers meet and establish an enduring relationship that also later serves, in deuterostomes, as the anatomical site of the anus. In the sea urchin, a prototypic deuterostome, the ectoderm-endoderm boundary is established before gastrulation, and ectodermal cells at the boundary are thought to provide patterning inputs to the underlying mesenchyme. Here we show that a short-range Wnt5 signal from the endoderm actively patterns the adjacent boundary ectoderm. This signal activates a unique subcircuit of the ectoderm gene regulatory network, including the transcription factors IrxA, Nk1, Pax2/5/8 and Lim1, which are ultimately restricted to subregions of the border ectoderm (BE). Surprisingly, Nodal and BMP2/4, previously shown to be activators of ectodermal specification and the secondary embryonic axis, instead restrict the expression of these genes to subregions of the BE. A detailed examination showed that endodermal Wnt5 functions as a short-range signal that activates only a narrow band of ectodermal cells, even though all ectoderm is competent to receive the signal. Thus, cells in the BE integrate positive and negative signals from both the primary and secondary embryonic axes to correctly locate and specify the border ectoderm. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
44. Morphogenesis in sea urchin embryos: linking cellular events to gene regulatory network states.
- Author
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Lyons, Deirdre C., Kaltenbach, Stacy L., and McClay, David R.
- Published
- 2012
- Full Text
- View/download PDF
45. Evolutionary crossroads in developmental biology: sea urchins.
- Author
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McClay, David R.
- Subjects
- *
DEVELOPMENTAL biology , *SEA urchins , *LIFE sciences , *EMBRYOS , *STARFISHES , *CELL lines - Abstract
Embryos of the echinoderms, especially those of sea urchins and sea stars, have been studied as model organisms for over 100 years. The simplicity of their early development, and the ease of experimentally perturbing this development, provides an excellent platform for mechanistic studies of cell specification and morphogenesis. As a result, echinoderms have contributed significantly to our understanding of many developmental mechanisms, including those that govern the structure and design of gene regulatory networks, those that direct cell lineage specification, and those that regulate the dynamic morphogenetic events that shape the early embryo. [ABSTRACT FROM AUTHOR]
- Published
- 2011
- Full Text
- View/download PDF
46. Dynamics of Delta/Notch signaling on endomesoderm segregation in the sea urchin embryo.
- Author
-
Croce, Jenifer C. and McClay, David R.
- Subjects
- *
MESODERM , *EMBRYONIC physiology , *VETERINARY embryology , *SEA urchin physiology , *CELL proliferation , *TRANSCRIPTION factors - Abstract
Endomesoderm is the common progenitor of endoderm and mesoderm early in the development of many animals. In the sea urchin embryo, the Delta/Notch pathway is necessary for the diversification of this tissue, as are two early transcription factors, Gcm and FoxA, which are expressed in mesoderm and endoderm, respectively. Here, we provide a detailed lineage analysis of the cleavages leading to endomesoderm segregation, and examine the expression patterns and the regulatory relationships of three known regulators of this cell fate dichotomy in the context of the lineages. We observed that endomesoderm segregation first occurs at hatched blastula stage. Prior to this stage, Gcm and FoxA are co-expressed in the same cells, whereas at hatching these genes are detected in two distinct cell populations. Gcm remains expressed in the most vegetal endomesoderm descendant cells, while FoxA is downregulated in those cells and activated in the above neighboring cells. Initially, Delta is expressed exclusively in the micromeres, where it is necessary for the most vegetal endomesoderm cell descendants to express Gcm and become mesoderm. Our experiments show a requirement for a continuous Delta input for more than two cleavages (or about 2.5 hours) before Gcm expression continues in those cells independently of further Delta input. Thus, this study provides new insights into the timing mechanisms and the molecular dynamics of endomesoderm segregation during sea urchin embryogenesis and into the mode of action of the Delta/Notch pathway in mediating mesoderm fate. [ABSTRACT FROM AUTHOR]
- Published
- 2010
- Full Text
- View/download PDF
47. Blocking Dishevelled signaling in the noncanonical Wnt pathway in sea urchins disrupts endoderm formation and spiculogenesis, but not secondary mesoderm formation.
- Author
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Byrum, Christine A., Xu, Ronghui, Bince, Joanna M., McClay, David R., and Wikramanayake, Athula H.
- Abstract
Dishevelled (Dsh) is a phosphoprotein key to beta-catenin dependent (canonical) and beta-catenin independent (noncanonical) Wnt signaling. Whereas canonical Wnt signaling has been intensively studied in sea urchin development, little is known about other Wnt pathways. To examine roles of these beta-catenin independent pathways in embryogenesis, we used Dsh-DEP, a deletion construct blocking planar cell polarity (PCP) and Wnt/Ca
2+ signaling. Embryos overexpressing Dsh-DEP failed to gastrulate or undergo skeletogenesis, but produced pigment cells. Although early mesodermal gene expression was largely unperturbed, embryos exhibited reduced expression of genes regulating endoderm specification and differentiation. Overexpressing activated beta-catenin failed to rescue Dsh-DEP embryos, indicating that Dsh-DEP blocks endoderm formation downstream of initial canonical Wnt signaling. Because Dsh-DEP-like constructs block PCP signaling in other metazoans, and disrupting RhoA or Fz 5/8 in echinoids blocks subsets of the Dsh-DEP phenotypes, our data suggest that noncanonical Wnt signaling is crucial for sea urchin endoderm formation and skeletogenesis. Developmental Dynamics 238:1649-1665, 2009. © 2009 Wiley-Liss, Inc. [ABSTRACT FROM AUTHOR]- Published
- 2009
- Full Text
- View/download PDF
48. Ingression of primary mesenchyme cells of the sea urchin embryo: A precisely timed epithelial mesenchymal transition.
- Author
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Wu, Shu-Yu, Ferkowicz, Michael, and McClay, David R.
- Published
- 2007
- Full Text
- View/download PDF
49. p38 MAPK in Development and Cancer.
- Author
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Bradham, Cynthia and McClay, David R.
- Published
- 2006
- Full Text
- View/download PDF
50. The canonical Wnt pathway in embryonic axis polarity
- Author
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Croce, Jenifer C. and McClay, David R.
- Subjects
- *
SEA urchins , *CELL polarity , *PIPIDAE , *TISSUES - Abstract
Abstract: The canonical Wnt pathway plays crucial roles in multiple developmental processes, including in axis specification. Throughout the animal kingdom, this pathway has been reported to drive patterning of axes as different as the animal–vegetal axis in echinoderms to the dorsal–ventral axis in vertebrates. Intriguingly enough, this pathway appears structurally and functionally well conserved during evolution. However, differences between these phyla are observed that explain how a same pathway can mediate establishment of two such apparently distinct axes. This review compares the axis specification processes used in two evolutionarily distant embryos, the sea urchin and Xenopus. [Copyright &y& Elsevier]
- Published
- 2006
- Full Text
- View/download PDF
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