Leech Segmental Repeats Develop Normally in the Absence of Signals from either Anterior or Posterior Segments

AbstractWe have investigated whether the development of segmental repeats is autonomous in the embryo of the leech Helobdella robusta. The segmental tissues of the germinal band arise from progeny of five stem cells called teloblasts. Asymmetric divisions of the teloblasts form chains of segment founder cells (called primary blast cells) that divide in a stereotypical manner to produce differentiated descendants. Using two distinct techniques, we have looked for potential interactions between neighboring blast cell clones along the anterior–posterior axis. In one technique, we prevented the birth of primary blast cells by injection of DNase I into the teloblast, thereby depriving the last blast cell produced before the ablation of its normal posterior neighbors. We also ablated single blast cells with a laser microbeam, which allowed us to assess potential signals acting on either more anterior or more posterior primary blast cell clones. Our results suggest that interactions along the anterior–posterior axis between neighboring primary blast cell clones are not required for development of normal segmental organization within the blast cell clone. We also examined the possibility that blast cells receive redundant signals from both anterior and posterior neighboring clones and that either is sufficient for normal development. Using double blast cell laser ablations to isolate a primary blast cell clone by removal of both its anterior and its posterior neighbor, we found that the isolated clone still develops normally. These results reveal that the fundamental segmental repeat in the leech embryo, the primary blast cell clone, can develop normally in the apparent absence of signals from adjacent repeats along the anterior–posterior axis

Expression of FoxA and GATA transcription factors correlates with regionalized gut development in two lophotrochozoan marine worms: Chaetopterus (Annelida) and Themiste lageniformis (Sipuncula)

<p>Abstract</p> <p>Background</p> <p>A through gut is present in almost all metazoans, and most likely represents an ancient innovation that enabled bilaterian animals to exploit a wide range of habitats. Molecular developmental studies indicate that <it>Fox </it>and <it>GATA </it>regulatory genes specify tissue regions along the gut tube in a broad diversity of taxa, although little is known about gut regionalization within the Lophotrochozoa. In this study, we isolated <it>FoxA </it>and <it>GATA456 </it>orthologs and used whole mount <it>in situ </it>hybridization during larval gut formation in two marine worms: the segmented, polychaete annelid <it>Chaetopterus</it>, which develops a planktotrophic larva with a tripartite gut, and the non-segmented sipunculan <it>Themiste lageniformis</it>, which develops a lecithotrophic larva with a U-shaped gut.</p> <p>Results</p> <p><it>FoxA </it>and <it>GATA456 </it>transcripts are predominantly restricted to gut tissue, and together show regional expression spanning most of the alimentary canal in each of these lophotrochozoans, although neither <it>FoxA </it>nor <it>GATA456 </it>is expressed in the posterior intestine of <it>Chaetopterus</it>. In both species, <it>FoxA </it>is expressed at the blastula stage, transiently in presumptive endoderm before formation of a definitive gut tube, and throughout early larval development in discrete foregut and hindgut domains. <it>GATA456 </it>genes are expressed during endoderm formation, and in endoderm and mesoderm associated with the midgut in each species. Several species-specific differences were detected, including an overlap of <it>FoxA </it>and <it>GATA456 </it>expression in the intestinal system of <it>Themiste</it>, which is instead complimentary in <it>Chaetopterus</it>. Other differences include additional discrete expression domains of <it>FoxA </it>in ectodermal trunk cells in <it>Themiste </it>but not <it>Chaetopterus</it>, and expression of <it>GATA456 </it>in anterior ectoderm and midgut cells unique to <it>Chaetopterus</it>.</p> <p>Conclusions</p> <p>This study of gene expression in a sipunculan contributes new comparative developmental insights from lophotrochozoans, and shows that <it>FoxA </it>and <it>GATA456 </it>transcription factors are part of an ancient patterning mechanism that was deployed during early evolution of the metazoan through gut. The common utilization of <it>FoxA </it>and <it>GATA456 </it>throughout gut formation by species with contrasting life history modes indicates that both genes are core components of a gut-specific gene regulatory network in spiralians. Despite a highly conserved pattern of early development, and probably similar ontogenic origins of gut tissue, there are molecular differences in gut regionalization between lophotrochozoan species.</p

Expression of the pair-rule gene homologs runt, Pax3/7, even-skipped-1 and even-skipped-2 during larval and juvenile development of the polychaete annelid Capitella teleta does not support a role in segmentation

<p>Abstract</p> <p>Background</p> <p>Annelids and arthropods each possess a segmented body. Whether this similarity represents an evolutionary convergence or inheritance from a common segmented ancestor is the subject of ongoing investigation.</p> <p>Methods</p> <p>To investigate whether annelids and arthropods share molecular components that control segmentation, we isolated orthologs of the <it>Drosophila melanogaster </it>pair-rule genes, <it>runt</it>, <it>paired </it>(<it>Pax3/7</it>) and <it>eve</it>, from the polychaete annelid <it>Capitella teleta </it>and used whole mount <it>in situ </it>hybridization to characterize their expression patterns.</p> <p>Results</p> <p>When segments first appear, expression of the single <it>C. teleta runt </it>ortholog is only detected in the brain. Later, <it>Ct-runt </it>is expressed in the ventral nerve cord, foregut and hindgut. Analysis of <it>Pax </it>genes in the <it>C. teleta </it>genome reveals the presence of a single <it>Pax3/7 </it>ortholog. <it>Ct-Pax3/7 </it>is initially detected in the mid-body prior to segmentation, but is restricted to two longitudinal bands in the ventral ectoderm. Each of the two <it>C. teleta eve </it>orthologs has a unique and complex expression pattern, although there is partial overlap in several tissues. Prior to and during segment formation, <it>Ct-eve1 </it>and <it>Ct-eve2 </it>are both expressed in the bilaterial pair of mesoteloblasts, while <it>Ct-eve1 </it>is expressed in the descendant mesodermal band cells. At later stages, <it>Ct-eve2 </it>is expressed in the central and peripheral nervous system, and in mesoderm along the dorsal midline. In late stage larvae and adults, <it>Ct-eve1 </it>and <it>Ct-eve2 </it>are expressed in the posterior growth zone.</p> <p>Conclusions</p> <p><it>C. teleta eve, Pax3/7 </it>and <it>runt </it>homologs all have distinct expression patterns and share expression domains with homologs from other bilaterians. None of the pair-rule orthologs examined in <it>C. teleta </it>exhibit segmental or pair-rule stripes of expression in the ectoderm or mesoderm, consistent with an independent origin of segmentation between annelids and arthropods.</p

A comprehensive fate map by intracellular injection of identified blastomeres in the marine polychaete Capitella teleta

<p>Abstract</p> <p>Background</p> <p>The polychaete annelid <it>Capitella teleta </it>(formerly <it>Capitella </it>sp. I) develops by spiral cleavage and has been the focus of several recent developmental studies aided by a fully sequenced genome. Fate mapping in polychaetes has lagged behind other spiralian taxa, because of technical limitations.</p> <p>Results</p> <p>To generate a modern fate map for <it>C. teleta</it>, we injected 1,1'-dioctadecyl-3,3,3'3'-tetramethylindocarbocyanine perchlorate (DiI) into individual identified blastomeres through fourth-quartet micromere formation. Confocal laser scanning microscopy at single-cell resolution was used to characterize blastomere fates during larval stages. Our results corroborate previous observations from classic studies, and show a number of similarities with other spiralian fate maps, including unique and stereotypic fates for individual blastomeres, presence of four discrete body domains arising from the A, B, C and D cell quadrants, generation of anterior ectoderm from first quartet micromeres, and contributions to trunk ectoderm and ventral nerve cord by the 2d somatoblast. Of particular interest are several instances in which the <it>C. teleta </it>fate map deviates from other spiralian fate maps. For example, we identified four to seven distinct origins of mesoderm, all ectomesodermal. In addition, the left and right mesodermal bands arise from 3d and 3c, respectively, whereas 4d generates a small number of trunk muscle cells, the primordial germ cells and the anus. We identified a complex set of blastomere contributions to the posterior gut in <it>C. teleta</it>, which establishes the most complete map of posterior gut territories to date.</p> <p>Conclusions</p> <p>Our detailed cellular descriptions reveal previously underappreciated complexity in the ontogenetic contributions to several spiralian larval tissues, including the mesoderm, nervous system and gut. The formation of the mesodermal bands by 3c and 3d is in stark contrast to other spiralians, in which 4d generates the mesodermal bands. The results of this study provide a framework for future phylogenetic comparisons and functional analyses of cell-fate specification.</p

Expression and phylogenetic analysis of the zic gene family in the evolution and development of metazoans

<p>Abstract</p> <p>Background</p> <p><it>zic </it>genes are members of the <it>gli/glis/nkl/zic </it>super-family of C2H2 zinc finger (ZF) transcription factors. Homologs of the <it>zic </it>family have been implicated in patterning neural and mesodermal tissues in bilaterians. Prior to this study, the origin of the metazoan <it>zic </it>gene family was unknown and expression of <it>zic </it>gene homologs during the development of early branching metazoans had not been investigated.</p> <p>Results</p> <p>Phylogenetic analyses of novel <it>zic </it>candidate genes identified a definitive <it>zic </it>homolog in the placozoan <it>Trichoplax adhaerens</it>, two <it>gli/glis/nkl-</it>like genes in the ctenophore <it>Mnemiopsis leidyi</it>, confirmed the presence of three <it>gli/glis/nkl</it>-like genes in Porifera, and confirmed the five previously identified <it>zic </it>genes in the cnidarian <it>Nematostella vectensis</it>. In the cnidarian <it>N. vectensis</it>, <it>zic </it>homologs are expressed in ectoderm and the gastrodermis (a bifunctional endomesoderm), in presumptive and developing tentacles, and in oral and sensory apical tuft ectoderm. The <it>Capitella teleta zic </it>homolog (<it>Ct-zic</it>) is detectable in a subset of the developing nervous system, the foregut, and the mesoderm associated with the segmentally repeated chaetae. Lastly, expression of <it>gli </it>and <it>glis </it>homologs in <it>Mnemiopsis</it>. <it>leidyi </it>is detected exclusively in neural cells in floor of the apical organ.</p> <p>Conclusions</p> <p>Based on our analyses, we propose that the <it>zic </it>gene family arose in the common ancestor of the Placozoa, Cnidaria and Bilateria from a <it>gli/glis/nkl</it>-like gene and that both ZOC and ZF-NC domains evolved prior to cnidarian-bilaterian divergence. We also conclude that <it>zic </it>expression in neural ectoderm and developing neurons is pervasive throughout the Metazoa and likely evolved from neural expression of an ancestral <it>gli/glis/nkl/zic </it>gene. <it>zic </it>expression in bilaterian mesoderm may be related to the expression in the gastrodermis of a cnidarian-bilaterian common ancestor.</p

Genomic Organization and Expression Demonstrate Spatial and Temporal Hox Gene Colinearity in the Lophotrochozoan Capitella sp. I

Hox genes define regional identities along the anterior–posterior axis in many animals. In a number of species, Hox genes are clustered in the genome, and the relative order of genes corresponds with position of expression in the body. Previous Hox gene studies in lophotrochozoans have reported expression for only a subset of the Hox gene complement and/or lack detailed genomic organization information, limiting interpretations of spatial and temporal colinearity in this diverse animal clade. We studied expression and genomic organization of the single Hox gene complement in the segmented polychaete annelid Capitella sp. I. Total genome searches identified 11 Hox genes in Capitella, representing 11 distinct paralog groups thought to represent the ancestral lophotrochozoan complement. At least 8 of the 11 Capitella Hox genes are genomically linked in a single cluster, have the same transcriptional orientation, and lack interspersed non-Hox genes. Studying their expression by situ hybridization, we find that the 11 Capitella Hox genes generally exhibit spatial and temporal colinearity. With the exception of CapI-Post1, Capitella Hox genes are all expressed in broad ectodermal domains during larval development, consistent with providing positional information along the anterior–posterior axis. The anterior genes CapI-lab, CapI-pb, and CapI-Hox3 initiate expression prior to the appearance of segments, while more posterior genes appear at or soon after segments appear. Many of the Capitella Hox genes have either an anterior or posterior expression boundary coinciding with the thoracic–abdomen transition, a major body tagma boundary. Following metamorphosis, several expression patterns change, including appearance of distinct posterior boundaries and restriction to the central nervous system. Capitella Hox genes have maintained a clustered organization, are expressed in the canonical anterior–posterior order found in other metazoans, and exhibit spatial and temporal colinearity, reflecting Hox gene characteristics that likely existed in the protostome–deuterostome ancestor

Developmental expression of COE across the Metazoa supports a conserved role in neuronal cell-type specification and mesodermal development

The transcription factor COE (collier/olfactory-1/early B cell factor) is an unusual basic helix–loop–helix transcription factor as it lacks a basic domain and is maintained as a single copy gene in the genomes of all currently analysed non-vertebrate Metazoan genomes. Given the unique features of the COE gene, its proposed ancestral role in the specification of chemosensory neurons and the wealth of functional data from vertebrates and Drosophila, the evolutionary history of the COE gene can be readily investigated. We have examined the ways in which COE expression has diversified among the Metazoa by analysing its expression from representatives of four disparate invertebrate phyla: Ctenophora (Mnemiopsis leidyi); Mollusca (Haliotis asinina); Annelida (Capitella teleta and Chaetopterus) and Echinodermata (Strongylocentrotus purpuratus). In addition, we have studied COE function with knockdown experiments in S. purpuratus, which indicate that COE is likely to be involved in repressing serotonergic cell fate in the apical ganglion of dipleurula larvae. These analyses suggest that COE has played an important role in the evolution of ectodermally derived tissues (likely primarily nervous tissues) and mesodermally derived tissues. Our results provide a broad evolutionary foundation from which further studies aimed at the functional characterisation and evolution of COE can be investigated

Annelids shed light on the evolution of spiralian development

Spiralian development is characterized by stereotypic cell geometry and spindle orientation in early cleavage-stage embryos, and of ultimate fates of descendent clones. Diverse taxa such as molluscs, annelids, flatworms and nemerteans exhibit spiralian development, but it is a mystery how such a conserved developmental program gives rise to such diverse body plans. This review highlights examples of variation during early development among spiralians, emphasizing recent experimental studies in the annelid Capitella teleta(Blake, 2009). Intracellular fate mapping studies in C. teleta reveal that many of its cellsâ fates are shared among spiralians, but it also has a previously undescribed novel origin for trunk mesoderm (3c and 3d micromeres). Studies have identified an inductive signal in spiralians that has â organizing activityâ , and influences cell fates in the surrounding embryo. C. teleta also has an organizing activity, however, surprisingly, it is localized to a different cell, it signals at a different developmental stage, and likely utilizes a distinct molecular signaling pathway compared with that in molluscs. A model is presented to provide a mechanistic explanation of evolutionary changes in the cellular identity of the organizer. Detailed experimental investigations in spiralian embryos demonstrate variation in developmental features that may influence the evolution of novel forms.The accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author

Regeneration in the Segmented Annelid Capitella teleta

The segmented worms, or annelids, are a clade within the Lophotrochozoa, one of the three bilaterian superclades. Annelids have long been models for regeneration studies due to their impressive regenerative abilities. Furthermore, the group exhibits variation in adult regeneration abilities with some species able to replace anterior segments, posterior segments, both or neither. Successful regeneration includes regrowth of complex organ systems, including the centralized nervous system, gut, musculature, nephridia and gonads. Here, regenerative capabilities of the annelid Capitella teleta are reviewed. C. teleta exhibits robust posterior regeneration and benefits from having an available sequenced genome and functional genomic tools available to study the molecular and cellular control of the regeneration response. The highly stereotypic developmental program of C. teleta provides opportunities to study adult regeneration and generate robust comparisons between development and regeneration