Enteropneust Research Articles

Theoretical Study on Chemiluminescence of H2O2-dependent Tetrachloro-1,4-benzoquinone.

Compared with O2-dependent chemiluminescence (CL) and bioluminescence (BL), H2O2-dependent ones are far from being completely understood. A two-step mechanism for H2O2-dependent CL production from tetrachloro-1,4-benzoquinone (TCBQ) was proposed based on experimental evidence of detecting the formation of several intermediates and products. This mechanism is not yet supported by theoretical evidence, and its details remain unknown. In the present paper, we performed multireference and (time-dependent) density functional theory calculations on the complete reaction process of TCBQ with H2O2 to produce CL. The calculations reproduced the experimentally observed two-step CL. Although the reactants are different, the first and second CLs follow very similar reaction processes and mechanisms. First, an anionic dioxetane is formed via five sequential reactions. The intrinsically produced •OH is crucial for forming dioxetane. Subsequently, the anionic dioxetane decomposes to produce an anionic excited-state (S1) product. A conical interaction of the ground and the S1-state potential energy surfaces is responsible for producing the S1-state product. Finally, the S1-state anionic product changes to its neutral form, and the latter emits light as an actual light emitter. This mechanism could be extended to luminescent systems of all H2O2-dependent tetrahalogenated quinoids, including acorn worms, because TCBQ/H2O2 is a typical representative of these luminescent systems.

Unexpected molecular and morphological diversity of hemichordate larvae from the Neotropics

AbstractThe diversity of tropical marine invertebrates is poorly documented, especially those groups for which collecting adults is difficult. We collected the planktonic tornaria larvae of hemichordates (acorn worms) to assess their hidden diversity in the Neotropics. Larvae were retrieved in plankton tows from waters of the Pacific and Caribbean coasts of Panama, followed by DNA barcoding of mitochondrial cytochrome c oxidase subunit I (COI) and 16S ribosomal DNA to estimate their diversity in the region. With moderate sampling efforts, we discovered six operational taxonomic units (OTUs) in the Bay of Panama on the Pacific coast, in contrast to the single species previously recorded for the entire Tropical Eastern Pacific. We found eight OTUs in Bocas del Toro province on the Caribbean coast, compared to seven species documented from adults in the entire Caribbean. All OTUs differed from each other and from named acorn worm sequences in GenBank by >10% pairwise distance in COI and >2% in 16S. Two of our OTUs matched 16S hemichordate sequences in GenBank: one was an unidentified or unnamed Balanoglossus from the Caribbean of Panama, and the other was an unidentified ptychoderid larva from the Bahamas. The species accumulation curves suggest that nearly all the species have been collected and only one more species might still remain undetected in the Pacific. In contrast, the Caribbean species accumulation curve suggests that further sampling could yield more than 10 additional OTUs. Tornaria from the 14 OTUs exhibited typical planktotrophic morphologies, and, in some cases, may be distinguished by differences in pigmentation and by the number of telotrochal ciliary bands, but in general, few diagnostic differences were detected.

Neurotransmitter Receptor Antagonism Inhibits the Sea Urchin Righting Response

The deuterostome superphylum comprises the chordates, which includes all vertebrates, echinoderms (including sea urchins and sea stars), and hemichordates (acorn worms). Despite the close phylogenetic relationship between invertebrate echinoderms and vertebrates, the understanding of echinoderm neurobiology lags far behind that of many other invertebrate species. To investigate molecular components of echinoderm behavior, we used the righting response, in which an inverted sea urchin will use its tube feet and spines to right itself, as a behavioral assay. Using purple sea urchins (Strongylocentrotus purpuratus) we demonstrate that immersion in drug solution is a simple and effective drug administration route. Using the drug immersion procedure, we show that the sea urchin righting response is inhibited in a dose‐dependent manner by the nicotinic acetylcholine receptor antagonist hexamethonium and also affected by the GABAA receptor antagonist bicuculline. In addition, decreasing the concentration of extracellular Ca2+ also inhibited the righting response. These data start to elucidate the receptor systems involved the control of sea urchin motor activity.Support or Funding InformationFunding was provided by the University of the South Conduff Fund and the University of the South Kresge Fund.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Cloning and characterization of a novel aspartate/glutamate racemase from the acorn worm Saccoglossus kowalevskii.

Previously, we demonstrated that the animal aspartate racemase (AspR) gene has evolved from the serine racemase (SerR) gene by acquisition of three consecutive serine residues (Ser155-Ser156-Ser157) involved in the strong AspR activity, and this event has occurred independently and frequently during animal evolution. In the present study, we cloned and characterized two mammalian SerR homologous genes from the hemichordate acorn worm (Saccoglossus kowalevskii). The enzymes have been identified as an AspR and an aspartate/glutamate racemase (Asp/GluR) on the basis of their kinetic parameters. The S. kowalevskii Asp/GluR shows comparable substrate affinity and high catalytic efficiency (kcat/Km) for both aspartate and glutamate and is the first reported enzyme from animals that can synthesize d-glutamate. Amino acid sequence alignment analysis and site-directed mutagenesis studies have revealed that the amino acid residue at position 156, which is serine in AspR and alanine in Asp/GluR, is associated with binding and recognition of glutamate and aspartate. Phylogenetic analysis suggests that the S. kowalevskii AspR gene has evolved from the SerR gene after the divergence of hemichordata and vertebrate lineages by acquisition of the three serine residues at position 155 to 157 as in the case of other animal AspR genes. Furthermore, the S. kowalevskii Asp/GluR gene is the result of AspR gene duplication and several amino acid substitutions including that of the 156th serine residue with alanine. The fact that SerR has acquired substrate specificity towards aspartate or glutamate raises the possibility that synthesis of other d-amino acids is carried out by enzymes evolved from SerR.

Arms of larval seastars of Pisaster ochraceus provide versatility in muscular and ciliary swimming.

Larval swimming with cilia, unaided by muscles, is the presumed ancestral condition for echinoderms, but use of muscles in swimming has evolved several times. Ciliation and musculature of the arms of brachiolaria-stage larvae in the family Asteriidae provide unusual versatility in the use of muscles in swimming. The muscles affect swimming in two different ways. (1) Contraction of muscles moves the arms, propelling the larva. (2) Contraction of muscles changes orientation of the arms, thereby changing direction of ciliary currents and direction of swimming. New observations of the brachiolaria of the asteriid seastar Pisaster ochraceus demonstrate more versatility in both of these uses of muscles than had been previously described: the posterolateral arms stroke in more ways to propel the larva forward and to change the direction of swimming, and more pairs of the arms point ciliary currents in more directions for changes in direction of swimming. Morphology of brachiolariae suggests that these uses of muscles in swimming evolved before divergence of the families Stichasteridae and Asteriidae within forcipulate asteroids. This versatile use of muscles for swimming, both alone and in combination with ciliary currents, further distinguishes the swimming of these brachiolariae from swimming by larvae of other echinoderms and larvae of acorn worms in the sister phylum Hemichordata.

Regeneration of the acorn worm pygochord with the implication for its convergent evolution with the notochord.

The origin of the notochord is a central issue in chordate evolution. This study examined the development of the acorn worm pygochord, a putative homologue of the notochord. Because the pygochord differentiates only after metamorphosis, the developmental was followed process by inducing regeneration after artificial amputation in Ptychodera flava. It was found that although the regeneration of the posterior part of the body did not proceed via formation of an obvious regeneration bud, pygochord regeneration was observed within a few weeks, possibly via trans-differentiation of endoderm cells. The expression of the fibrillary collagen gene (Fcol) and elav in the pygochord during regeneration was detected. This indicates that pygochord cells are not part of gut epithelial cells, but that they differentiated as a distinct cell type. Our gene expression analyses do not provide supporting evidence for the homology between the pygochord and notochord, but rather favored the convergent evolution between them.

Biogeography and adaptations of torquaratorid acorn worms (Hemichordata: Enteropneusta) including two new species from the Canadian Arctic

The enteropneust family Torquaratoridae, discovered in 2005, has the fewest species of the four living families. It is composed of seven species that live on the cold, deep-sea floor. Torquarator bullocki Holland, Clague, Gordon, Gebruk, Pawson and Vecchione, 2005 was the first species described and collected from the Northeastern Pacific. Two new species of Torquaratoridae were collected from the Eastern Pacific and described as Tergivelum baldwinae Holland, Jones, Ellena, Ruhl and Smith, 2009 and Allapasus aurantiacus Holland, Kuhnz and Osborn, 2012. A further three species have been collected from the North Atlantic: Yoda purpurata Priede, Osborn, Gebruk, Jones, Shale, Rogacheva and Holland, 2012, Allapasus isidis Priede, Osborn, Gebruk, Jones, Shale, Rogacheva and Holland, 2012, and Tergivelum cinnabarinum Priede, Osborn, Gebruk, Jones, Shale, Rogacheva and Holland, 2012. The latest Torquaratoridae species was collected from the Russian Arctic in 2013. We add two new species to the Torquaratoridae from Baffin Bay and Viscount Melville Sound, in the eastern Arctic of Canada. Terminstomo arcticus gen. nov. and sp. nov. was collected at a depth of 505 m and is characterized by lacking a heart and a stomochord that extends from the posterior end of the proboscis through the entire length of the collar. Allapasus fuscus sp. nov. was collected from a depth of 444 m and is characterized by a typhlosole process along the dorsal midline of the collar buccal cavity.

Regeneration in the enteropneust hemichordate, Ptychodera flava, and its evolutionary implications.

Hemichordates are marine invertebrates that are closely related to chordates, but while their body plans are comparable to those of chordates, they possess a remarkable capacity for regeneration, even as adults. A small fragment is sufficient to form a complete individual. Unlike echinoderms, their larvae transform directly into adults; therefore, hemichordate systems offer clear morphological and molecular parallels between regeneration and development. Morphological events in regeneration are generally similar to organogenesis in juveniles. Nonetheless, comparative analysis of gene expression in these two morphological phenomena suggests that hemichordate regeneration is regulated by regeneration-specific mechanisms, as well as by developmental mechanisms. Dependency upon resident pluripotent/multipotent stem cells is a significant difference in metazoan regeneration, and such stem cells are essential for regeneration in many lineages. Based on the present gene expression study, regeneration in acorn worms is more closely related to that in vertebrates, because it employs endogenous stem cell-independent transdifferentiation.

About the Cover

Next article FreeAbout the CoverPDFPDF PLUSFull Text Add to favoritesDownload CitationTrack CitationsPermissionsReprints Share onFacebookTwitterLinked InRedditEmailQR Code SectionsMoreCoverThe image on the cover is a Ptychodera flava hemichordate in the Krohn stage of larval development. This larva will undergo a dramatic metamorphosis to become a tripartite, marine acorn worm. The nuclei are stained with DAPI (blue), and dividing cells are stained with the marker EdU (green). The ciliary bands that wrap around the animal are densely populated with cells and can be seen as rich blue, wavy lines, a prominent character of the Krohn stage. Significant cell proliferation is also apparent throughout the ciliary bands and C-shaped collar and trunk coeloms sprouting from the midgut region.Ptychodera flava larvae undergo a long larval stage and can drift in the plankton for approximately a year before undergoing metamorphosis. The larval body must develop though several progressive developmental stages before becoming competent for metamorphosis.On pages 152–164, S. M. Luttrell, Y.-H. Su, and B. J. Swalla show that within two to four weeks after bisection through the sagittal, coronal, or axial planes, P. flava larvae completely regenerate all structures. Regeneration fidelity is not equal between the different larval halves and between the different bisection planes. This suggests that an unknown stem cell niche may be required for complete regeneration. EdU staining was performed to determine whether dividing cells are bona fide stem cells. While adult P. flava acorn worms are known to regenerate all tissues, this report provides new knowledge about regeneration in P. flava during the larval stage of development.Credits: Photo, Shawn Luttrell, University of Washington. Cover design, Jeannie Harrell, University of Chicago Press. Next article DetailsFiguresReferencesCited by The Biological Bulletin Volume 234, Number 3June 2018 Published in association with the Marine Biological Laboratory Article DOIhttps://doi.org/10.1086/699540 © 2018 by The University of Chicago. All rights reserved.PDF download Crossref reports no articles citing this article.

Getting a Head with Ptychodera flava Larval Regeneration

Severe injury to the central nervous system of chordates often results in permanent and irreversible mental and physical challenges. While some chordates are able to repair and/or regenerate portions of their nervous system, no chordate has been shown to be able to regenerate all regions of its central nervous system after catastrophic injury or amputation. Some hemichordates, on the other hand, are able to efficiently regenerate all neural structures, including their dorsal, hollow neural tube after complete ablation. Solitary hemichordates are marine acorn worms and a sister group to the echinoderms. The hemichordate Ptychodera flava progresses from a pelagic, feeding tornaria larva to a tripartite benthic worm with an anterior proboscis, a middle collar region, and a long posterior trunk. The adult worm regenerates all body parts when bisected in the trunk, but it was unknown whether the regeneration process was present in tornaria larvae. Now, we show that P. flava larvae are capable of robust regeneration after bisection through the sagittal, coronal, and axial planes. We also use antibody staining to show that the apical sensory organ regenerates a rich, serotonin-positive complex of cells within two weeks after amputation. Cells labeled with 5-ethynyl-2'-deoxyuridine confirm that regeneration is occurring through epimorphic processes as new cells are added at the cut site and throughout the regenerating tissue. This study verifies that P. flava larvae can be used for future functional studies aimed at identifying the genetic and morphological mechanisms controlling central nervous system regeneration in a stem deuterostome.

Horizontal transfer of retrotransposons between bivalves and other aquatic species of multiple phyla

The LTR retrotransposon Steamer is a selfish endogenous element in the soft-shell clam genome that was first detected because of its dramatic amplification in bivalve transmissible neoplasia afflicting the species. We amplified and sequenced related retrotransposons from the genomic DNA of many other bivalve species, finding evidence of horizontal transfer of retrotransposons from the genome of one species to another. First, the phylogenetic tree of the Steamer-like elements from 19 bivalve species is markedly discordant with host phylogeny, suggesting frequent cross-species transfer throughout bivalve evolution. Second, sequences nearly identical to Steamer were identified in the genomes of Atlantic razor clams and Baltic clams, indicating recent transfer. Finally, a search of the National Center for Biotechnology Information sequence database revealed that Steamer-like elements are present in the genomes of completely unrelated organisms, including zebrafish, sea urchin, acorn worms, and coral. Phylogenetic incongruity, a patchy distribution, and a higher similarity than would be expected by vertical inheritance all provide evidence for multiple long-distance cross-phyla horizontal transfer events. These data suggest that over both short- and long-term evolutionary timescales, Steamer-like retrotransposons, much like retroviruses, can move between organisms and integrate new copies into new host genomes.

Selected Least Studied but not Forgotten Bioluminescent Systems.

Bioluminescence is a form of chemiluminescence generated by luminous organisms. Luminous taxa have currently been reported from about 800 genera and probably over 10 000 species in the world. On the other hand, their bioluminescent systems, including chemical structures of luciferins/chromophores and the genes encoding luciferases/photoproteins, have been elucidated from only a few taxonomic groups, for example beetles, bacteria, dinoflagellates, ostracods and some cnidarians. Research efforts to understand unknown bioluminescence systems are being conducted around the world, and recently, for example, novel luciferin structures of luminous enchytraeid potworms and fungi were identified by the authors. In this study, we review the current status and perspectives, in the context of postgenomic era, of most likely novel but less-revealed bioluminescence systems of ten selected organisms: earthworm, parchment tubeworm, fireworm, scaleworm, limpet, millipede, brittle star, acorn worms, tunicate and shark, which indeed are the next focus of our international collaboration.

Larval Evolution: I’ll Tail You Later…

Larval stages can be astonishingly different from their adult forms. A new study in acorn worms shows that the whole larval body is patterned only with a subset of anterior genes, revealing the intricate developmental bases that underlie the evolution of larval forms.

Head regeneration in hemichordates is not a strict recapitulation of development.

Background: Head or anterior body part regeneration is commonly associated with protostome, but not deuterostome invertebrates. However, it has been shown that the solitary hemichordate Ptychodera flava possesses the remarkable capacity to regenerate their entire nervous system, including their dorsal neural tube and their anterior head‐like structure, or proboscis. Hemichordates, also known as acorn worms, are marine invertebrate deuterostomes that have retained chordate traits that were likely present in the deuterostome ancestor, placing these animals in a vital position to study regeneration and chordate evolution. All acorn worms have a tripartite body plan, with an anterior proboscis, middle collar region, and a posterior trunk. The collar houses a hollow, dorsal neural tube in ptychoderid hemichordates and numerous chordate genes involved in brain and spinal cord development are expressed in a similar anterior–posterior spatial arrangement along the body axis. Results: We have examined anterior regeneration in the hemichordate Ptychodera flava and report the spatial and temporal morphological changes that occur. Additionally, we have sequenced, assembled, and analyzed the transcriptome for eight stages of regenerating P. flava, revealing significant differential gene expression between regenerating and control animals. Conclusions: Importantly, we have uncovered developmental steps that are regeneration‐specific and do not strictly follow the embryonic program. Developmental Dynamics 245:1159–1175, 2016. © 2016 The Authors. Developmental Dynamics published by Wiley Periodicals, Inc. on behalf of American Association of Anatomists

The Global Diversity of Hemichordata

Phylum Hemichordata, composed of worm-like Enteropneusta and colonial Pterobranchia, has been reported to only contain about 100 species. However, recent studies of hemichordate phylogeny and taxonomy suggest the species number has been largely underestimated. One issue is that species must be described by experts, and historically few taxonomists have studied this group of marine invertebrates. Despite this previous lack of coverage, interest in hemichordates has piqued in the past couple of decades, as they are critical to understanding the evolution of chordates–as acorn worms likely resemble the deuterostome ancestor more closely than any other extant animal. This review provides an overview of our current knowledge of hemichordates, focusing specifically on their global biodiversity, geographic distribution, and taxonomy. Using information available in the World Register of Marine Species and published literature, we assembled a list of 130 described, extant species. The majority (83%) of these species are enteropneusts, and more taxonomic descriptions are forthcoming. Ptychoderidae contained the greatest number of species (41 species), closely followed by Harrimaniidae (40 species), of the recognized hemichordate families. Hemichordates are found throughout the world’s oceans, with the highest reported numbers by regions with marine labs and diligent taxonomic efforts (e.g. North Pacific and North Atlantic). Pterobranchs are abundant in Antarctica, but have also been found at lower latitudes. We consider this a baseline report and expect new species of Hemichordata will continue to be discovered and described as new marine habitats are characterized and explored.

Characterization of paramyosin and thin filaments in the smooth muscle of acorn worm, a member of hemichordates.

Paramyosin is a myosin-binding protein characteristic of invertebrate animals, while troponin is a Ca2+-dependent regulator of muscle contraction. Both proteins are widely distributed in protostomes, while in deuterostomes, their distribution is limited; namely, presence of paramyosin and absence of troponin are common features in echinoderm muscles, while muscles of chordates contain troponin but lack paramyosin. In this study, we examined the muscle of a hemichordate, acorn worm, to clarify whether this animal is like echinoderms or like the other deuterostome animals. We found a 100-kDa protein in the smooth muscle of acorn worm. This protein was identified with paramyosin, since the purified protein formed paracrystals with a constant axial periodicity in the presence of divalent cations as paramyosin of other animals, showed ability to interact with myosin and shared common antigenicity with echinoderm paramyosin. On the other hand, troponin band was not detected in isolated thin filaments, and the filaments increased myosin-ATPase activity in a Ca2+-independent manner. The results indicate that troponin is lacking in thin filaments of acorn worm muscle just as in those of echinoderms. The muscle of hemichordate acorn worm is quite similar to echinoderm muscles, but different from chordate muscles.

Brachiopods, sipunculans, enteropneusts, and metals from two estuarine tidal flats, Pacific, Costa Rica

Reports on the abundances and on metal concentrations in intertidal estuarine invertebrates from the Eastern Tropical Pacific are rare. Thus, the objectives of this report are to make accessible data on the abundances (1984-1987, 49 dates; 2013, 12 dates) of sipunculans, brachiopods and hemichordates from a sand-mud flat; and on trace metals (1996, 2000) and abundances (2015, 3 dates) of sipunculans and brachiopods at a sand flat in the Gulf of Nicoya estuary (10o N-85o W). Cores (17.7 cm2) were collected at the sand-mud flat, and quadrats (0.2 m2) at the sand flat. The flats contrasted in their sand (65 % vs 90 %) and silt+clay (31.5 % vs 5.6 %) contents. At the sand-mud flat (1984-87: 1.83 m2) the sipunculans were represented by 13 individuals, the brachiopods by 129 and the acorn worms by 185, with estimated maximum densities of: 5.7, 29, and 40 ind./m2, respectively. Trace metal (Fe, Mn, Ni, Cr, Cd, Zn, and Pb) analysis (Atomic Absorption Spectrometry) were conducted in specimens of Sipunculus nudus (Sipuncula) and Glottidia audebarti (Brachiopoda). Maximum mean concentrations in S. nudus were: For non-depurated worms, Fe (16.0 mg/g dw) > Mn (165 μg/g dw) > Zn (81 μg/g dw) > Cu (26 μg/g dw) > Cr (11 μg/g dw) > Ni (10.4 μg/g dw) > Pb (9.3 μg/g dw) > Cd (1.2 μg/g dw). For 72 hour depurated worms: Fe (5.0 mg/g dw) > Mn (61 μg/g dw) > Zn (39 μg/g dw) > Cu (24 μg/g dw) > Ni (8.4 μg/g dw) > Pb (2.7 μg/g dw) > Cd (0.62 μg/g dw). For G. audebarti: Fe (1.6 mg/g dw-soft parts) > Zn (123.5 μg/g dw-soft parts) > Cu (31.4 μg/g dw-pedicles) > Pb (21.0 μg/g dw-shells) > Cd (5.2 μg/g dw-soft parts) > Cr (4.7 μg/g dw-shells). For sediments; Fe (46 mg/g dw) > Mn (41.3 μg/g dw) > Zn (63 μg/g dw) > Cu (36.2 μg/g dw) > Cr (31.5 μg/g dw) > Pb (21.1 μg/g dw) > Ni (16.1 μg/g dw) > Cd (1.1 μg/g dw). These concentrations were expected for a nonindustrialized estuary. At the sand flat (Area sampled: 10.6 m2 ) 76 individuals of G. audebarti, 112 of G. albida, and 366 of S. nudus were collected in 2015, with estimated maximum densities of: 7.1, 10.5, and 31 ind./m2, respectively. Densities of G. audebarti and G. albida were relatively low, while those of S. nudus were relatively high when compared with other reports. The shell lenght of G. audebarti ranged from 9.0 mm to 38.0 mm and from 6.0 mm to 29.0 mm for G. albida. These ranges were within those found for these lingulides elsewhere. The mean length of S. nudus was 41 mm and the maximum weight was 1.6 g, which are small. No brachiopods were found at the sand-mud flat in 2013, nor enteropneusts at the sand flat in 2015. G. audebarti had a relatively stable presence, while G. albida almost vanished from the samples at the end of 2015. The spatial distributions of the three invertebrates were found aggregated at both intertidal flats. Strong ENSO warming events during 1983 and 2015, and red tides in 1985 may have influenced the abundances.

Saccoglossus testafrom the Mazon Creek fauna (Pennsylvanian of Illinois) and the evolution of acorn worms (Enteropneusta: Hemichordata)

The limited fossil record of enteropneust hemichordates (acorn worms) and the few external features that distinguish the four families have provided a challenge to our understanding of the evolution of the group and their various feeding adaptations. The middle Pennsylvanian Saccoglossus testa sp. nov. from the Mazon Creek, Westfalian D Carbonate Formation, Francis Creek Shale of northern Illinois provides evidence for the exploitation of surface sediments. Saccoglossus testa has a long proboscis characteristic of the extant genus Saccoglossus, a specialist in surface deposit feeding. The collar is as long as it is wide. The anterior trunk lacks a distinctively wide branchial region. These three features distinguish it from its sympatric enteropneust species Mazoglossus ramsdelli Bardack that has a proboscis characteristic of an infaunal deposit feeder. It is the seventh known species of fossil enteropneust, including a resting trace of a Lower Triassic fossil that has collar lips that characterize the extant deep-sea family Torquaratoridae, and which represents a second parallel evolution of surface deposit feeding. An analysis of the seven fossils shows that the earliest Enteropneusta had a relatively simple harrimaniid-like body plan, and that the spengelid, the torquaratorid and lastly the most complex ptychoderid body plan appeared in that chronological order.

Reproductive periodicity, spawning induction, and larval metamorphosis of the hemichordate acorn worm Ptychodera flava.

The indirect-developing enteropneust acorn worm Ptychodera flava has been used as a hemichordate model system for studying the developmental evolution of deuterostome body plans and the origins of chordate characteristics. However, research progress has been hindered by the limited accessibility of its embryonic materials and metamorphosing larvae. In this study, we identified an abundant population of P. flava in Penghu, Taiwan, and examined the feasibility of using this animal for developmental studies. Through histological examination, we established that the reproductive season of this population is between September and December, with a peak breeding period in October and November. In addition, we have developed new procedures that can induce P. flava spawning at any time of the day during the breeding season, with a higher successful rate than that achieved using a previously published method. Moreover, the culturing system we developed enables rearing of P. flava larvae through various planktonic stages and eventual metamorphosis into benthic juveniles, all under laboratory conditions. We anticipate that the animal resources and new technical procedures reported here will further facilitate the use of P. flava as a model organism for evolutionary and developmental biology research.

FGF signaling repertoire of the indirect developing hemichordate Ptychodera flava

Fibroblast growth factors (FGFs) are a group of ligands that play multiple roles during development by transducing signals through FGF receptors (FGFRs) to downstream factors. At least 22 FGF ligands and 4 receptors have been identified in vertebrates, while six to eight FGF ligands and a single FGFR are present in invertebrate chordates, such as tunicates and amphioxus. The chordate FGFs can be categorized into at least seven subfamilies, and the members of which expanded during the evolution of early vertebrates. In contrast, only one FGF and two FGFRs have been found in sea urchins. Thus, it is unclear whether the FGF subfamilies duplicated in the lineage leading to the chordates, or sea urchins lost several fgf genes. Analyses of the FGF signaling repertoire in hemichordates, which together with echinoderms form the closest group to the chordates, may provide insights into the evolution of FGF signaling in deuterostomes. In this study, we identified five FGFs and three FGFRs from Ptychodera flava, an indirect-developing hemichordate acorn worm. Phylogenetic analyses revealed that hemichordates possess a conserved FGF8/17/18 in addition to several putative hemichordate-specific FGFs. Analyses of sequence similarity and protein domain organizations suggested that the sea urchin and hemichordate FGFRs arose from independent lineage-specific duplications. Furthermore, the acorn worm fgf and fgfr genes were demonstrated to be expressed during P. flava embryogenesis. These results set the foundations for further functional studies of FGF signaling in hemichordates and provided insights into the evolutionary history of the FGF repertoire.