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Appendicularians are planktonic tunicates abundant all over the world. Currently, only two complete annotated mitochondrial genome assemblies are available for appendicularians, both for cryptic species of Oikopleura dioica. This underrepresentation of available appendicularian mitochondrial genomes limits environmental DNA sequencing (eDNA) studies that rely on mitochondrial markers as a taxonomic barcode. We report the complete mitochondrial genome assembly and annotation of an unknown appendicularian species isolated from the Amami Oshima island, Kagoshima prefecture, Japan, that has significant sequence difference with other currently available assemblies and will serve as a useful resource for ecological studies and further mitochondrial studies of appendicularians.

Appendicularian tunicates are some of the most abundant mesozooplankton organisms with key roles in marine trophic webs and global carbon flux. Like most appendicularians with cosmopolitan distributions, Oikopleura dioica Fol, 1872 is considered a single species worldwide based on morphological features that distinguish them from other appendicularians. Despite their abundance, however, there are still only ~ 70 described appendicularian species, compared to over 2800 ascidian tunicates. Here we perform a molecular phylogenetic, morphological, and reproductive assessment of O. dioica specimens collected from the Ryukyu Archipelago, mainland Japan, and Europe. The specimens are morphologically very similar, with only detailed examination of the oikoplastic epithelium and quantitative measurements revealing minor distinguishing characteristics. Phylogenetic analyses of the ribosomal gene loci and mitochondrial cytochrome oxidase I (COI) gene strongly indicate that they form three separate genetic clades despite their morphological similarities. Finally, in vitro crosses between the Ryukyu and mainland Japanese specimens show total prezygotic reproductive isolation. Our results reveal that the current taxonomic O. dioica classification likely hides multiple cryptic species, highlighting the genetic diversity and complexity of their population structures. Cryptic organisms are often hidden under a single species name because their morphological similarities make them difficult to distinguish and their correct identification is fundamental to understanding Earth's biodiversity. O. dioica is an attractive model to understand how morphological conservation can be maintained despite pronounced genetic divergence. [ABSTRACT FROM AUTHOR]

Inuit are at the forefront of ecosystem change in the Arctic, yet their observations and interpretations are rarely reported in the literature. Climate change impacts are rapidly unfolding in the Arctic and there is a need for monitoring and reporting unique observations. In this short communication, we draw upon observations and experiential knowledge from western Canadian Inuit (Inuvialuit) harvesters combined with a scientific assessment to describe and interpret an unusual account of gelatinous organisms at high densities during summer 2019 in eastern Amundsen Gulf, near Ulukhaktok, Northwest Territories. The gelatinous organisms were identified as primarily appendicularian larvaceans (Oikopleura spp., pelagic tunicates) and their gelatinous “houses”. The organisms were observed within 3–5 km of the marine coast, from ∼1–2 m below the surface and to depths of ∼30 m with an underwater camera. Pelagic tunicates have rarely been documented in the eastern Amundsen Gulf and, to our knowledge, this was the first time these organisms had been noted by the people of Ulukhaktok. The pelagic tunicates clogged subsistence fishing nets and Inuvialuit harvesters were concerned about negative impacts to marine mammals and fishes, which they depend on for food security. These interpretations highlight major knowledge gaps for appendicularians in the Arctic.Video Inuit Ukiuktaktumi nunamingni tautukpaktut nunaktik aalangnujuhianik, taimaa tautukpaktait ilihimaliktait titiraqtauyuitut titiqani. Nunam aallangujuhia tautuktauyuq kayumikhipluni Ukiuktaktun nunanni, taimatun munariyauyukhak titiraqhimayukhat aallangujuhiit. Uvani tittiqaniInuit tautukpaktait ilihimaliktait titiraqhimayut Inuvialuit anguniaqtiinnit attauttimut iliblugit qablunaat tittiratainnutilituritiarumaplugitumayuutigut tamainnut auyanani 2019mi, tahamani Admundson Gulfmi Ulukhaktuum haniani North west Territoriesmi. Tahapkuat uumayut hauniittut imangmi attauttimiitpaktut. Uumayut tahapkuat tautuktauvaktut pingahunikluunniin tallimanitulluunniin kilometresmik ungahiktilanganik tariukmitalvattauk atauhikmikluunniin, malguknikluunniin metresnik ititigiyumi. Ilaani ititqiyami 30 metresmi takunnaqpaktun ajiliurunmun. Tahapkuninga tautuyuittugaluat taja kihimi tautukpaliktait. Ulukhaktuunmiutat taja tautukpaliktait. Tahapkuat uumayut nuvaktut inmate kuvyanut himiutkpiaqpaktut, kuvyallu iqaluguikhutik. Inuit iqalukhiuktullu anguniaqtullu ihumaalugiliktait anguyamingnik niqiqaramik. Tahapkuat ilituriliktavut ilihimatttaingitnaptigit nunaptingni ukiuktaktumi ilitturrinahuaqqaqhaluavut.

Appendicularians are the most important group among zooplankton communities, widely distributed in all the oceans and they are considered as one of the most important sources of productivity and food supplies for secondary consumers. So this study aimed to identify, introduce and assess the temporal and spatial variations of the Appendicularians in the Persian Gulf, restricted to coastal waters of northern part of Bushehr. Appendicularians were collected during seven sampling programs, from Frake river-estuary, three sites of large creek-estuary of the Shif, one site in the sea, and one site in the Lashkari creek-estuary, at middle and last day of each season, midsummer of 2016 to spring of 2017. During all surveys, depth of the sampling location, Secchi depth, water temperature, pH, and salinity were measured. Finally, 4 species were identified, Oikopleura dioica, Oikopleura longicauda Appendicularia sicula and Fritillaria sp. The highest total density was observed in the sea station at March, with dominance of O. dioica and a relative abundance of 80%, relative abundance of O. longicauda, A sicula andFritillaria sp.were calculated 18.9%, 1.06% and 0.04% respectively, so Fritillaria sp were introduced as rare species. Two species of O. dioica andO. longicauda demonstrated completely different spatial and temporal distribution pattern, and based on dbRDA (distance based Redundancy Analysis), transparency explains effectively the distribution pattern of O longicauda, and alsodepth and salinity are the most important factors which explain the distribution pattern of O dioica and temperature was correlated factor with abundances of A sicula, in our study area.

We examined the feeding habits of the dominant mesopelagic fish Sigmops gracilis larvae across the Kuroshio Current, southern Japan. The feeding incidence tended to decrease toward the offshore area. Copepods, amphipods, ostracods, and appendicularians were microscopically observed in the guts. Larvae fed substantially on copepods such as calanoids and appendicularians. Appendicularians were more frequently ingested toward the offshore area and were the most preferential prey item in all areas. The microbial production integrated by appendicularians would support mesopelagic fish larvae in addition to the ordinary trophic pathway through copepods. [ABSTRACT FROM AUTHOR]

Unfertilized eggs of most animals are arrested at a certain point in the meiotic cell cycles. Reinitiation of meiosis and the start of embryogenesis are triggered by fertilization. This arrest is essential for preventing parthenogenetic activation and for promoting proper initiation of development by fertilization. In the larvacean Oikopleura dioica , which is a simple model organism for studies of chordate development, the unfertilized egg is arrested at metaphase of meiosis I. We show here that protein phosphatase 2A (PP2A) is essential for maintenance of meiotic arrest after spawning of oocytes. Knockdown (KD) of the maternal PP2A catalytic subunit, which was found in functional screening of maternal factors, caused unfertilized eggs to spontaneously release polar bodies after spawning, and then start pseudo-cleavages without fertilization, namely, parthenogenesis. Parthenogenetic embryos failed to undergo proper mitosis and cytokinesis because of lack of a centrosome, which is to be brought into the egg by a sperm. Activation of the KD oocytes was triggered by possible rise of ambient and intracellular pH upon their release from the gonad into seawater at spawning. Live recording of intracellular calcium level of the KD oocytes indicated that the pH rise caused an aberrant Ca2+ burst, which mimicked the Ca2+ burst that occurs at fertilization. Then, the aberrant Ca2+ burst triggered meiosis resumption through Calcium/calmodulin-dependent protein kinase (CaMK II). Therefore, PP2A is essential for maintenance of meiotic arrest and prevention of parthenogenesis by suppressing the aberrant Ca2+ burst at spawning. • PP2A is essential for maintenance of meiotic arrest in O. dioica oocytes. • Knock down of PP2Ac resulted in parthenogenesis after spawning. • Ambient pH rise at spawning promoted aberrant Ca2+ burst in PP2Ac-KD oocytes. • The aberrant Ca2+ burst led oocytes to reinitiate meiosis and pseudo-cleavages via CaMK II. • O. dioica ' s mechanism of meiotic arrest is more similar to that of vertebrates than ascidians. [ABSTRACT FROM AUTHOR]

Abstract We characterized spontaneous Ca2+ signals in Oikopleura dioica embryos from pre-fertilization to gastrula stages following injection of GCaMP6 mRNA into unfertilized eggs. The unfertilized egg exhibited regular, transient elevations in intracellular Ca2+ concentration with an average duration of 4–6 s and an average frequency of about 1 every 2.5 min. Fertilization was accompanied by a longer Ca2+ transient that lasted several minutes. Thereafter, regular Ca2+ transients were reinstated that spread within seconds among blastomeres and gradually increased in duration (by about 50%) and decreased in frequency (by about 20%) by gastrulation. Peak amplitudes also exhibited a dynamic, with a transitory drop occurring at about the 4-cell stage and a subsequent rise. Each peak was preceded by about 15 s by a smaller and shorter Ca2+ increase (about 5% of the main peak amplitude, average duration 3 s), which we term the "minipeak". By gastrulation, Ca2+ transients exhibited a stereotyped initiation site on either side of the 32-64-cell embryo, likely in the nascent muscle precursor cells, and spread thereafter symmetrically in a stereotyped spatial pattern that engaged blastomeres giving rise to all the major tissue lineages. The rapid spread of the transients relative to the intertransient interval created a coordinated wave that, on a coarse time scale, could be considered an approximate synchronization. Treatment with the divalent cations Ni2+ or Cd2+ gradually diminished peak amplitudes, had only moderate effects on wave frequency, but markedly disrupted wave synchronization and normal development. The T-type Ca2+ channel blocker mibefradil similarly disrupted normal development, and eliminated the minipeaks, but did not affect wave synchronization. To assess the role of gap junctions in calcium wave spread and coordination, we first characterized the expression of two Oikopleura connexins, Od-CxA and Od-CxB, both of which are expressed during pre-gastrulation and gastrula stages, and then co-injected double-stranded inhibitory RNAs together with CGaMP6 to suppress connexin expression. Connexin mRNA knockdown led to a gradual increase in Ca2+ transient peak width, a decrease of interpeak interval and a marked disruption of wave synchronization. As seen with divalent cations and mibefradil, this desynchronization was accompanied by a disruption of normal development. Highlights • Spontaneous, regular calcium signals appear in the unfertilized oocyte. • Cleavage through gastrulation, evolve into coordinated intercellular calcium waves. • Calcium waves are initiated consistently in muscle lineage precursors. • Calcium channel blockers disrupt wave coordination and embryonic development. [ABSTRACT FROM AUTHOR]

The appendicularian Oikopleura dioica is a planktonic chordate that retains a tadpole shape throughout its life. Its simple and transparent body, invariant cell lineages, fast development and available genome and transcriptome resources make it a promising model organism for research in developmental biology. However, large-scale analysis of gene expression in O. dioica is limited owing to the laborious and time-consuming process of manual removal of the vitelline membrane, because devitellinisation of pre-hatching embryos causes failure of normal development. Therefore, in this study, modified procedures were developed for whole-mount in situ hybridisation (WISH) and immunohistochemistry (WIHC). This protocol enables rapid mRNA or protein detection without a manual devitellination step for each specimen. The critical procedure is brief treatment of the vitelline membrane of living embryos with 0.05% actinase E before fixation. Two minutes of treatment was optimal for the penetration of antisense RNA probes and antibodies through the vitelline membrane. This WISH protocol was applicable for chromogenic and fluorescent tyramide signal amplification reactions. Using the new protocol, we found eight genes with tissue-specific expression in the tail muscle, trunk epidermis, heart, pharynx, oesophagus, stomach or gill openings of developing larvae. This procedure also allowed for the detection of exogenous FLAG-tagged histone-enhanced green fluorescent protein by WIHC using anti-FLAG antibody. This study provides a useful and convenient tool for studying spatial and temporal gene expression patterns in this simple chordate model and should facilitate handling large amounts of genetic data from transcriptome-based approaches and other techniques such as treatments with chemical inhibitors. [ABSTRACT FROM AUTHOR]

Mechanisms for morphogenetic processes that generate complex patterns in a reproducible manner remain elusive. Live imaging provides a powerful tool to record cell behaviors. The appendicularian, Oikopleura dioica , is a planktonic tunicate that has a rapid developmental speed, small number of cells (less than 3500 cells in a juvenile), and a transparent body. The trunk epidermis, called the oikoplastic epithelium (OE), has elaborate cellular arrangements showing a complex pattern to secrete so-called “house” made of extracellular components. The OE is characterized by invariant number, size, and shape of the monolayer epithelial cells. Pattern formation is achieved during 5 h of larval development without growth of the body, making this a suitable system for live imaging of a two-dimensional (2D) sheet. First, we subdivided the OE and defined several domains by cellular resolution, and systematically gave names to the constituent cells, since there is no variation among individuals. Time-lapse imaging of the epidermal cells revealed region-specific pattern formation processes. Each identified domain served as a compartment into which distribution of descendant cells of founder cells is restricted. Regulation of orientation, timing, and the number of rounds of cell divisions, but not cell death and migration, was a critical mechanism for determination of final cell arrangement and size. In addition, displacement of epithelial sheet plates was observed in the Eisen domain. Stem-cell-like cell divisions, whereby large mother stem cells generate a chain of small daughter cells, were involved in formation of the Nasse region and ventral sensory organ. These are the first examples of this kind of stem-cell-like cell division in deuterostomes. Furthermore, labeling of the left or right blastomere of the two-cell-stage embryo, which roughly gives rise to the left or right side of the body, respectively, revealed that the boundary of the descendant cells does not match with the midline of the trunk epidermis. Left and right descendants largely invade into the opposite side in an invariant way, suggesting the possibility that specification of the OE cell identities may occur later in development, most probably around hatching, and depending on cell position in the OE epithelial sheet. These detailed descriptions of OE patterning processes provide basic and essential information to analyze further cell behaviors in the generation of elaborate and intricate but stereotyped 2D cellular patterns in this advantageous model system for developmental and cell biological studies in chordates. [ABSTRACT FROM AUTHOR]

The appendicularian, Oikopleura dioica, is a planktonic tunicate that retains a swimming tadpole shape throughout its life. It has relatively few cells and exhibits fast development, yet it has a basic chordate body plan. In this study, the morphology of adults was investigated using scanning electron microscopy (SEM) and fine 3D images of most organs were taken. The trunk epidermis is organized into bilateral territories secreting the house that includes the food-trapping filter. The pharynx extends ventrally and posteriorly to the gill openings and esophagus, respectively. The endostyle, with a morphologically distinct ciliated band, is embedded in the pharynx. The digestive tract showed left-right asymmetry as the connection between the pharynx and esophagus tilts leftward. The heart is located ventrally between the left stomach and the intestine and consists of a left muscular sheet and a right thin, non-muscular sheet. The brain is connected to the oral and ventral sensory organs, ciliary funnels and sensory vesicles and axons descend from it that eventually innervate the caudal ganglion. In the tail, a nerve cord with sporadically distributed neuronal somata runs along the left side of the notochord. The gonad is a single syncytium of thousands of gametes. In the ovary, an abundance of cortical membrane invaginate into the cytoplasm during oogenesis and the growing oocytes are interconnected via common cytoplasm through a ring canal. Spermatogenesis progresses synchronously within the common cytoplasm. These descriptions provide a valuable anatomical atlas for studying development and physiology using this simple organism with a chordate body plan. [ABSTRACT FROM AUTHOR]

Background: Larvaceans in the genus Bathochordaeus are large, often abundant filter feeders found throughout much of the world ocean. The first described species, Bathochordaeus charon, was reported over 100 years ago by Chun. However in the time since, few specimens have matched Chun's original description, resulting in ambiguity on the validity of B. charon as a species. Methods: Specimens of Bathochordaeus charon were identified based on morphological traits, molecular data and observations made on high definition video. Results: The first records of Bathochordaeus charon from the northeast Pacific Ocean off central California and Oregon, USA are reported. Morphology and molecular data clearly distinguish B. charon from its congener, B. stygius. Conclusions: This paper establishes the first review of Bathochordaeus charon since its original description, extends the range of this species to the northeast Pacific Ocean, and provides the first molecular evidence for two species of Bathochordaeus. [ABSTRACT FROM AUTHOR]

RNA sequencing analysis was carried out to characterize egg and larval transcriptomes in the appendicularian, Oikopleura dioica, a planktonic chordate, which is characterized by rapid development and short life cycle of 5 days, using a Japanese population of the organism. De novo transcriptome assembly matched with 16,423 proteins corresponding to 95.4 % of the protein-encoding genes deposited in the OikoBase, the genome database of the Norwegian population. Nucleotide and amino acid sequence identities between the Japanese and Norwegian O. dioica were estimated to be around 91.0 and 94.8 %, respectively. We discovered 175 novel protein-encoding genes: 144 unigenes were common to both the Japanese and Norwegian populations, whereas 31 unigenes were not found in the OikoBase genome reference. Among the total 12,311 unigenes, approximately 63 % were detected in egg-stage RNAs, whereas 99 % were detected in larval stage RNAs; 3772 genes were up-regulated, and 1336 genes were down-regulated more than four-fold in the larvae. Gene ontology analyses characterized gene activities in these two developmental stages. We found a messenger RNA (mRNA) 5′ trans-spliced leader, which was observed in 40.8 % of the total unique transcripts. It showed preferential linkage to adenine at the 5′ ends of the downstream exons. Trans-splicing was observed more frequently in egg mRNAs compared with larva-specific mRNAs. [ABSTRACT FROM AUTHOR]

Larvacean (=appendicularian) and pteropod (Limacina helicina) composition and abundance were studied with physical variables each May and late summer across 11 years (2001–2011), along a transect that crosses the continental shelf of the sub-Arctic Gulf of Alaska (GoA) and five stations within Prince William Sound (PWS). Collection with 53-µm plankton nets allowed the identification of larvaceans to species: five occurred in the study area. Temperature was the driving variable in determining larvacean community composition, yielding pronounced differences between spring and late summer, while individual species were also affected differentially by salinity and chlorophyll-a concentration. During the spring Oikopleura labradoriensis and Fritillaria borealis were most abundant, being present at all stations. Late summer had highest abundances of Oikopleura dioica at nearshore stations, while F. borealis dominated numerically at outer stations. The 53-µm plankton nets collected higher abundances of Oikopleura spp., Fritillaria spp. and L. helicina than coarser 150- and 505-µm plankton nets. Limacina helicina abundance had a significant interaction effect among years, seasons and station location. Limacina helicina abundance in nearby PWS explained 30% of the variability in pink salmon survival; however, no significant correlations existed with larvacean or L. helicina abundances from the GoA stations. [ABSTRACT FROM PUBLISHER]

The appendicularian, Oikopleura dioica , is a planktonic chordate. Its simple and transparent body, invariant cell lineages and short life cycle of 5 days make it a promising model organism for studies of chordate development. Here we describe the cell migration that occurs during development of the O. dioica larva. Using time-lapse imaging facilitated by florescent labeling of cells, three cell populations exhibiting long-distance migration were identified and characterized. These included (i) a multinucleated oral gland precursor that migrates anteriorly within the trunk region and eventually separates into the left and right sides, (ii) endodermal strand cells that are collectively retracted from the tail into the trunk in a tractor movement, and (iii) two subchordal cell precursors that individually migrate out from the trunk to the tip of the tail. The migration of subchordal cell precursors starts when all of the endodermal strand cells enter the trunk, and follows the same path but in a direction opposite to that of the latter. Labeling of these cells with a photoconvertible fluorescent protein, Kaede, demonstrated that the endodermal strand cells and subchordal cell precursors have distinct origins and eventual fates. Surgical removal of the trunk from the tail demonstrated that the endodermal strand cells do not require the trunk for migration, and that the subchordal cell precursors would be attracted by the distal part of the tail. This well-defined, invariant and traceable long-distance cell migration provides a unique experimental system for exploring the mechanisms of versatile cell migration in this simple organism with a chordate body plan. [ABSTRACT FROM AUTHOR]

The appendicularian Oikopleura dioica is a chordate that has a remarkably simple adult body with small cell number. Its transparency, stereotyped cell lineages, short life cycle, and small genome make it a promising new experimental model of chordate developmental biology. However, the functions of its various genes are still poorly understood due to lack of a tool for suppression of gene expression. Here, we applied a double-stranded RNA (dsRNA)-based-RNA interference (RNAi) method in O. dioica. For introducing dsRNA into eggs and embryos, we injected dsRNAs into the ovary. dsRNA, which is specific to EGFP or mCherry mRNA, decreased the exogenous mRNA-derived fluorescence in both eggs and embryos. dsRNA specific to the Brachyury gene of O. dioica, which is a homologous gene of a key notochord transcriptional factor in ascidians, triggered degradation of endogenous Brachyury mRNA and induced malformation or loss of the notochord in the tail. This effect was Brachyury sequence specific, as three dsRNAs covering different sequences produced the same phenotype. The result is in accordance with its expression site and also with the key regulatory function of Brachyury in notochord formation in other chordates. RNAi in O. dioica would be a useful tool for gaining insight into the oogenesis and early developmental processes in chordates. [ABSTRACT FROM AUTHOR]

Deibel, D., and Lowen, B. 2012. A review of the life cycles and life-history adaptations of pelagic tunicates to environmental conditions. – ICES Journal of Marine Science, 69: 358–369.Phylogeny, life cycles, and life-history adaptations of pelagic tunicates to temperature and food concentration are reviewed. Using literature data on lifetime egg production and generation time of appendicularians, salps, and doliolids, rmax, the maximum rate of lifetime reproductive fitness, is calculated as a common metric of adaptation to environmental conditions. The rmax values are high for all three groups, ranging from ∼0.1 to 1.9 d−1, so population doubling times range from ∼8 h to 1 week. These high values of rmax are attributable primarily to short generation times, ranging from 2 to 50 d. Clearly, pelagic tunicates are adapted to event-scale (i.e. days to weeks) rather than seasonal-scale changes in environmental conditions. Although they are not closely related phylogenetically, all three groups have a unique life-history adaptation promoting high lifetime fitness. Appendicularians have late oocyte selection, salps are viviparous, and doliolids possess a polymorphic asexual phase. There has been little research on hermaphroditic appendicularians, on large oceanic salps, and on doliolids generally. Research is needed on factors regulating generation time, on the heritability of life-history traits, and on age- and size-specific rates of mortality. [ABSTRACT FROM PUBLISHER]

So far, the role of appendicularians in the biogeochemical cycling of organic matter has been largely overlooked. Appendicularians represent only a fraction of total mesozooplankton biomass, however these ubiquitous zooplankters have very high filtration and growth rates compared to copepods, and produce numerous fecal pellets and filtering houses contributing to export production by aggregating small marine particles. To study their quantitative impact on biogeochemical flux, we have included this group in the biogeochemical flux model, using a recently developed ecophysiological model. One-dimensional annual simulations of the pelagic ecosystem including appendicularians were conducted with realistic surface forcing for the year 2000, using data from the DyFAMed open ocean station. The appendicularian grazing impact was generally low, but appendicularians increased detritus production by 8% and export production by 55% compared to a simulation without appendicularians. Therefore, current biogeochemical models lacking appendicularians probably under, or misestimate the detritus and export production by omitting the pathway from small-sized plankton to fast sinking detritus. Detritus production and export rates are 60% lower than the estimates from mesotrophic sites, showing that appendicularians’ role is lower but still significant in oligotrophic environments. The simulated annual export at 200 m exceeds sediment trap values by 44%, suggesting an intense degradation during the sinking of appendicularian detritus, supported by observations made at other sites. Thus, degradation and grazing of appendicularian detritus need better quantification if we are to accurately assess the role of appendicularia in export flux. [ABSTRACT FROM AUTHOR]

The native form of cellulose is a fibrillar composite of two crystalline phases, the triclinic I and monoclinic I allomorphs. Allomorph ratios are species-specific, and this gives rise to natural structural variations in cellulose crystals. However, the mechanisms contributing to crystal formation remain unknown. We show that the two crystalline phases of cellulose are tailored to distinct structures during different developmental stages of the tunicate chordate Oikopleura dioica. Larval cellulose consisting of I allomorph constitutes the body cuticle fin, whereas adult cellulose consisting of I allomorph frames a mucous filter-feeding device, the 'house.' Both structures are secreted from the epidermis in accordance with the mutually exclusive expression patterns of two distinct putative cellulose synthase genes. We discuss a possible linkage between structural variations of the crystalline phases of cellulose and the underlying evolutionary genetics of cellulose biosynthesis. [ABSTRACT FROM AUTHOR]

Abstract: A model has been developed to simulate the growth of an individual appendicularian (Oikopleura dioica) from egg to spawning. This model uses a new set of experimental data presented in a companion paper in this volume and estimates growth rates and generation times as well as clearance rates, house and fecal pellet production and reproductive capacity at different temperatures and food concentrations. According to the model outputs, the weight of a single house represents 11.5% of the individual''s total weight, a lower value than previously estimated. The relative weight of one fecal pellet varies as a function of food concentration. The model also confirms that the minimum food concentration for growth is about 20–30 µg C l−1 and that growth is maximal for a 100 µg C l−1 feeding concentration. The limits of the growth optimum in relation to food concentration and temperature can be considered as a first description of the fundamental ecological niche of appendicularians. This model can be used both for predicting (1) the impact of in situ observed populations on the pico- and nanoseston and (2) the production of large aggregates. These two pieces of information are often needed for large-scale biogeochemical models. [Copyright &y& Elsevier]

Abstract: Oogenesis in the tunicate, Oikopleura, is unusual for a chordate, in that the thousands of nuclei comprising the entire germline are contained in a unique giant cell, the coenocyst. We examined progression through meiotic prophase I in concert with cellular mechanisms implicated in selection, growth and maturation of oocytes in this shared cytoplasm. Unlike sister vertebrates, no germinal vesicle was formed and maternal transcripts were instead synthesized by polyploid nurse nuclei present in equal numbers to transcriptionally quiescent meiotic nuclei. Meiosis resumption was concomitant with MAPK cascade activation during which pERK translocated to nurse nuclei. Simultaneously, the coenocyst partitioned into hundreds of synchronously growing oocytes. Significantly, only the subset of meiotic nuclei selected to populate maturing oocytes displayed histone H3 serine 28 phosphorylation. Disruption of the MAPK cascade, or microtubule dynamics, did not inhibit meiotic resumption but generated oocytes with multiple nurse and meiotic nuclei. As these supernumerary nuclei also became H3S28P enriched, growing oocytes defined a selective kinase environment in the common coenocyst cytoplasm. Vitellogenesis preceded the timing of oocyte selection among excess germ line nuclei in contrast to Drosophila and vertebrates. This unique feature enables late adjustment of oocyte number in accordance with the cytoplasmic volume of the germline cyst accumulated during vitellogenesis. [Copyright &y& Elsevier]

Appendicularians are planktonic tunicates (urochordates), and retain a swimming tadpole shape throughout their life. Together with ascidians, they are the closest relatives of the vertebrates. Oikopleura dioica is characterized by its simplified life habit and anatomical organization. It has a tiny genome, the smallest ever found in a chordate. Its life cycle is extremely short – about 5 days – and it can be maintained in the laboratory over many generations. Embryos and adults are transparent and consist of a small number of cells. The anatomy of juveniles and adults has been described in detail. Cleavage pattern, cell lineages, and morphogenetic movements during embryogenesis have also been comprehensively documented. A draft genome sequence is now available. These features make this organism a suitable experimental model animal in which genetic manipulations would be feasible, as in Drosophila and Caenorhabditis elegans. In this review, I summarize a hundred years’ knowledge on the development throughout the life cycle of this organism. Oikopleura is an attractive organism for developmental and evolutionary studies of chordates. It offers considerable promise for future genetic approaches. [ABSTRACT FROM AUTHOR]

The appendicularian, Oikopleura dioica is a chordate. Its life cycle is extremely short—approximately 5 days—and its tadpole shape with a beating tail is retained throughout entire life. The tadpole hatches after 3 h of development at 20°C. Here, we describe the cleavage pattern and morphogenetic cell movements during gastrulation and neurulation. Cleavage showed an invariant pattern. It is basically bilateral but also shows various minor left–right asymmetries starting from the four-cell stage. We observed two rounds of unequal cleavage of the posterior-vegetal B-line cells at the posterior pole. The nature of the unequal cleavages is reminiscent of those in ascidian embryos and suggests the presence of a centrosome-attracting body, a special subcellular structure at the posterior pole. The representation of the cell division pattern in this report will aid the identification of each cell, a prerequisite for clarifying the gene expression patterns in early embryos. Gastrulation started as early as the 32-cell stage and progressed in three phases. By the end of the second phase at the 64-cell stage, every vegetal cell had ingressed into the embryo, and animal cells had covered the entire embryo by epiboly. There was no archenteron formation. In the anterior region, eight A-line cells were aligned as a 2 × 4 array along the anterior–posterior axis and become internalized during the 64-cell stage. This process was considered to correspond to neurulation. The simple and accelerated development of Oikopleura, nevertheless giving rise to a conserved chordate body plan, is advantageous for studying developmental mechanisms using molecular and genetic approaches and makes this animal the simplest model organism in the phylum Chordata. [ABSTRACT FROM AUTHOR]

Abstract: Metazoan lamins are implicated in the organization of numerous critical nuclear processes. Among chordates, the appendicularian, Oikopleura dioica, has an unusually short life cycle involving rapid growth through extensive recourse to endoreduplication, a characteristic more associated with some invertebrates. In some tissues, this is accompanied by the formation of elaborate, bilaterally symmetric nuclear morphologies associated with specific gene expression patterns. Lamin composition can mediate nuclear shape in spermiogenesis as well as during pathological and normal aging and we have analyzed the O. dioica lamin and intermediate filament (IF) complement, comparing it to that present in other deuterostomes. O. dioica has one lamin gene coding two splice variants. Both variants share with the sister class ascidians a highly reduced C-terminal tail region lacking the immunoglobulin fold, indicating this derivation occurred at the base of the urochordate lineage. The OdLamin2 variant has a unique insertion of 63 amino acids in the normally short N-terminal region and has a developmental expression profile corresponding to the occurrence of endocycling. O. dioica has 4 cytoplasmic IF proteins, IF-A, C, Dα, and Dβ. No homologues to the ascidian IF-B or F proteins were identified, reinforcing the suggestion that these proteins are unique to ascidians. The degree of sequence evolution in the rod domains of O. dioica cytoplasmic IF proteins and their closest ascidian and vertebrate homologues was similar. In contrast, whereas the rate of lamin B rod domain sequence evolution has also been similar in vertebrates, cephalochordates and the sea urchin, faster rates have occurred among the urochordates, with the O. dioica lamin displaying a far greater rate than any other lamin. [Copyright &y& Elsevier]

Abstract: Developmental signaling by retinoic acid (RA) is thought to be an innovation essential for the origin of the chordate body plan. The larvacean urochordate Oikopleura dioica maintains a chordate body plan throughout life, and yet its genome appears to lack genes for RA synthesis, degradation, and reception. This suggests the hypothesis that the RA-machinery was lost during larvacean evolution, and predicts that Oikopleura development has become independent of RA-signaling. This prediction raises the problem that the anterior–posterior organization of a chordate body plan can be developed without the classical morphogenetic role of RA. To address this problem, we performed pharmacological treatments and analyses of developmental molecular markers to investigate whether RA acts in anterior–posterior axial patterning in Oikopleura embryos. Results revealed that RA does not cause homeotic posteriorization in Oikopleura as it does in vertebrates and cephalochordates, and showed that a chordate can develop the phylotypic body plan in the absence of the classical morphogenetic role of RA. A comparison of Oikopleura and ascidian evidence suggests that the lack of RA-induced homeotic posteriorization is a shared derived feature of urochordates. We discuss possible relationships of altered roles of RA in urochordate development to genomic events, such as rupture of the Hox-cluster, in the context of a new understanding of chordate phylogeny. [Copyright &y& Elsevier]

To clarify the feeding strategy of pelagic larvae of stone flounder in Mutsu Bay, the dietary composition and prey size was investigated from February to April during 1989–1999. Diets were compared with the numerical and volumetric compositions and frequency of occurrence of each prey species. Mensuration formulae were applied to estimate individual prey volume in the diet, while the length of planktonic species was measured from net and water samples. Prey shapes were assumed as sphere, cylinder, ellipsoid, pyramid, two elliptical cones, or a combination of ellipsoid and cylinder. Prey-size range increased as the larvae grew. Preflexion larvae fed mainly on copepod nauplii. Flexion and postflexion larvae ingested primarily appendicularians, with a suggestion that these larvae might depend on some parts of the microbial food web. Low frequencies of flexion and postflexion larvae with empty guts (1.7 and 1.4%, respectively) might be derived from feeding on slow-swimming appendicularians. From a size comparison between ‘house’-like organ length and trunk length of the appendicularian Oikopleura sp., almost all house-like organs with trunks in the larval diet were nonexpanded ‘house rudiments’, not expanded ‘houses’. Thus, stone flounder larvae may not chew the houses, but swallow the house rudiments with trunks. [ABSTRACT FROM AUTHOR]

The feeding ecology of Pacific Bluefin tuna (PBT: Thunnus orientalis) larvae collected from two nursery grounds, namely the Japan Sea and the Nansei area (western North Pacific), was evaluated based on gut content analyses using morphological and metagenetic techniques. The PBT larvae were collected at the surface layer (≥30 m depth) in 2016 and 2017, and 172 and 114 individuals from the Japan Sea and Nansei area, respectively, were analyzed. A generalized additive model applied revealed that the number of prey in the gut increased with growth, in waters above 24 °C and in subsurface (~30 m depth) waters, during daytime. The total dry weights of prey per gut calculated from the size of copepods and cladocerans also increased during daytime and with growth. Both the morphological and metagenetic analyses indicated that the taxonomic composition of gut contents was influenced by the food selectivity of PBT larvae and by zooplankton (prey) availability in the ambient water. Podonidae were positively selected as PBT larvae prey in both areas, and copepodites and adult Corycaeidae and Pontellidae were positively selected in the Nansei area only. A metagenetic approach indicated Calanidae copepods and soft zooplankton such as appendicularians as important preys in both areas. Other copepods were not positively selected using this approach, but Clausocalanidae and Oithonidae were frequently detected in the Japan Sea. This might be related to the higher abundances of these families in the Japan Sea than in the Nansei area. On the other hand, the cladoceran Penilia avirostris and the copepods Echinostomatidae and Oncaeidae were rarely observed in the guts of PBT larvae although were abundant in the water column. Overall, PBT larvae feed abundantly and grow fast in warm and long-day waters rich in Podonidae, Corycaeidae, Pontellidae, and larval copepods, which contributes to their survival and recruitment in the Japan Sea and Nansei area. [ABSTRACT FROM AUTHOR]