Comparative genomics of mortal and immortal cnidarians unveils novel keys behind rejuvenation.

Turritopsis dohrnii is the only metazoan able to rejuvenate repeatedly after its medusae reproduce, hinting at biological immortality and challenging our understanding of aging. We present and compare whole-genome assemblies of T. dohrnii and the nonimmortal Turritopsis rubra using automatic and manual annotations, together with the transcriptome of life cycle reversal (LCR) process of T. dohrnii. We have identified variants and expansions of genes associated with replication, DNA repair, telomere maintenance, redox environment, stem cell population, and intercellular communication. Moreover, we have found silencing of polycomb repressive complex 2 targets and activation of pluripotency targets during LCR, which points to these transcription factors as pluripotency inducers in T. dohrnii. Accordingly, we propose these factors as key elements in the ability of T. dohrnii to undergo rejuvenation.

Apoptosis and cell proliferation during metamorphosis of the planula larva of Clytia hemisphaerica (Hydrozoa, Cnidaria).

BACKGROUND: Metamorphosis in marine species is characterized by profound changes at the ecophysiological, morphological, and cellular levels. The cnidarian Clytia hemisphaerica exhibits a triphasic life cycle that includes a planula larva, a colonial polyp, and a sexually reproductive medusa. Most studies so far have focused on the embryogenesis of this species, whereas its metamorphosis has been only partially studied. RESULTS: We investigated the main morphological changes of the planula larva of Clytia during the metamorphosis, and the associated cell proliferation and apoptosis. Based on our observations of planulae at successive times following artificial metamorphosis induction using GLWamide, we subdivided the Clytia's metamorphosis into a series of eight morphological stages occurring during a pre-settlement phase (from metamorphosis induction to planula ready for settlement) and the post-settlement phase (from planula settlement to primary polyp). Drastic morphological changes prior to definitive adhesion to the substrate were accompanied by specific patterns of stem-cell proliferation as well as apoptosis in both ectoderm and endoderm. Further waves of apoptosis occurring once the larva has settled were associated with morphogenesis of the primary polyp. CONCLUSION: Clytia larval metamorphosis is characterized by distinct patterns of apoptosis and cell proliferation during the pre-settlement phase and the settled planula-to-polyp transformation.

Diminished growth and vitality in juvenile Hydractinia echinata under anticipated future temperature and variable nutrient conditions.

In a warming climate, rising seawater temperatures and declining primary and secondary production will drastically affect growth and fitness of marine invertebrates in the northern Atlantic Ocean. To study the ecological performance of juvenile hydroids Hydractinia echinata we exposed them to current and predicted water temperatures which reflect the conditions in the inter- and subtidal in combination with changing food availability (high and low) in laboratory experiments. Here we show, that the interplay between temperature stress and diminished nutrition affected growth and vitality of juvenile hydroids more than either factor alone, while high food availability mitigated their stress responses. Our numerical growth model indicated that the growth of juvenile hydroids at temperatures beyond their optimum is a saturation function of energy availability. We demonstrated that the combined effects of environmental stressors should be taken into consideration when evaluating consequences of climate change. Interactive effects of ocean warming, decreasing resource availability and increasing organismal energy demand may have major impacts on biodiversity and ecosystem function.

The evolution and development of coloniality in hydrozoans.

Hydrozoan colonies display a variety of shapes and sizes including encrusting, upright, and pelagic forms. Phylogenetic patterns reveal a complex evolutionary history of these distinct colony forms, as well as colony loss. Within a species, phenotypic variation in colonies as a response to changing environmental cues and resources has been documented. The patterns of branching of colony specific tissue, called stolons in encrusting colonies and stalks in upright colonies, are likely under the control of signaling mechanisms whose changing expression in evolution and development are responsible for the diversity of hydrozoan colony forms. Although mechanisms of polyp development have been well studied, little research has focused on colony development and patterning. In the few studies that investigated mechanisms governing colony patterning, the Wnt signaling pathway has been implicated. The diversity of colony form, evolutionary patterns, and mechanisms of colony variation in Hydrozoa are reviewed here.

Arctic coastal benthos long-term responses to perturbations under climate warming.

Climate warming influences structure and function of Arctic benthic ecosystems. Assessing the response of these systems to perturbations requires long-term studies addressing key ecological processes related to recolonization and succession of species. Based on unique time-series (1980-2017), this study addresses successional patterns of hard-bottom benthos in two fjords in NW Svalbard after a pulse perturbation in 1980 and during a period of rapid climate warming. Analysis of seafloor photographs revealed different return rates of taxa, and variability in species densities, through time. It took 13 and 24 years for the community compositions of cleared and control transects to converge in the two fjords. Nearly two decades after the study initiation, an increase in filamentous and foliose macroalgae was observed with a subsequent reorganization in the invertebrate community. Trait analyses showed a decrease in body size and longevity of taxa in response to the pulse perturbation and a shift towards small/medium size and intermediate longevity following the macroalgae takeover. The observed slow recovery rates and abrupt shifts in community structure document the vulnerability of Arctic coastal ecosystems to perturbations and continued effects of climate warming. This article is part of the theme issue 'The changing Arctic Ocean: consequences for biological communities, biogeochemical processes and ecosystem functioning'.

An improved whole life cycle culture protocol for the hydrozoan genetic model Clytia hemisphaerica.

The jellyfish species Clytia hemisphaerica (Cnidaria, Hydrozoa) has emerged as a new experimental model animal in the last decade. Favorable characteristics include a fully transparent body suitable for microscopy, daily gamete production and a relatively short life cycle. Furthermore, whole genome sequence assembly and efficient gene editing techniques using CRISPR/Cas9 have opened new possibilities for genetic studies. The quasi-immortal vegetatively-growing polyp colony stage provides a practical means to maintain mutant strains. In the context of developing Clytia as a genetic model, we report here an improved whole life cycle culture method including an aquarium tank system designed for culture of the tiny jellyfish form. We have compared different feeding regimes using Artemia larvae as food and demonstrate that the stage-dependent feeding control is the key for rapid and reliable medusa and polyp rearing. Metamorphosis of the planula larvae into a polyp colony can be induced efficiently using a new synthetic peptide. The optimized procedures detailed here make it practical to generate genetically modified Clytia strains and to maintain their whole life cycle in the laboratory.This article has an associated First Person interview with the two first authors of the paper.

Genetic examination of the marine bacterium Pseudoalteromonas luteoviolacea and effects of its metamorphosis-inducing factors.

Pseudoalteromonas luteoviolacea is a globally distributed marine bacterium that stimulates the metamorphosis of marine animal larvae, an important bacteria-animal interaction that can promote the recruitment of animals to benthic ecosystems. Recently, different P. luteoviolacea isolates have been shown to produce two stimulatory factors that can induce tubeworm and coral metamorphosis; Metamorphosis-Associated Contractile structures (MACs) and tetrabromopyrrole (TBP) respectively. However, it remains unclear what proportion of P. luteoviolacea isolates possess the genes encoding MACs, and what phenotypic effect MACs and TBP have on other larval species. Here, we show that 9 of 19 sequenced P. luteoviolacea genomes genetically encode both MACs and TBP. While P. luteoviolacea biofilms producing MACs stimulate the metamorphosis of the tubeworm Hydroides elegans, TBP biosynthesis genes had no effect under the conditions tested. Although MACs are lethal to larvae of the cnidarian Hydractinia symbiologicarpus, P. luteoviolacea mutants unable to produce MACs are capable of stimulating metamorphosis. Our findings reveal a hidden complexity of interactions between a single bacterial species, the factors it produces and two species of larvae belonging to different phyla.

Morphology and development of the Portuguese man of war, Physalia physalis.

The Portuguese man of war, Physalia physalis, is one of the most conspicuous, but poorly understood members of the pleuston, a community of organisms that occupy a habitat at the sea-air interface. Physalia physalis is a siphonophore that uses a gas-filled float as a sail to catch the wind. The development, morphology, and colony organization of P. physalis is very different from all other siphonophores. Here, we look at live and fixed larval and juvenile specimens, and use optical projection tomography to build on existing knowledge about the morphology and development of this species. We also propose a framework for homologizing the axes with other siphonophores, and also suggest that the tentacle bearing zooids should be called tentacular palpons. Previous descriptions of P. physalis larvae, especially descriptions of budding order, were often framed with the mature colony in mind. However, we use the simpler organization of larvae and the juvenile specimens to inform our understanding of the morphology, budding order, and colony organization in the mature specimen. Finally, we review what is known about the ecology and lifecycle of P. physalis.

Diversity and life-cycle analysis of Pacific Ocean zooplankton by videomicroscopy and DNA barcoding: Hydrozoa.

Most, but not all cnidarian species in the class Hydrozoa have a life cycle in which a colonial, asexually reproducing hydroid phase alternates with a free-swimming, sexually reproducing medusa phase. They are not well known, in part because many of them are microscopic, at least in the medusa phase. Matching the two phases has previously required rearing of the organism from one phase to another, which has not often been possible. Here we show that DNA barcoding makes it possible to easily link life-cycle phases without the need for laboratory rearing. Hydrozoan medusae were collected by zooplankton tows in Newport Bay and the Pacific Ocean near Newport Beach, California, and hydroid colonies were collected from solid substrates in the same areas. Specimens were documented by videomicroscopy, preserved in ethanol, and sent to the Canadian Centre for DNA Barcoding at the University of Guelph, Ontario, Canada for sequencing of the COI DNA barcode. In the order Anthoathecata (athecate hydroids), DNA barcoding allowed for the discrimination between the medusae of eight putative species of Bougainvillia, and the hydroid stages were documented for two of these. The medusae of three putative species of Amphinema were identified, and the hydroid stages were identified for two of them. DNA barcodes were obtained from medusae of one species of Cladonema, one adult of the by-the wind Sailor, Velella velella, five putative species of Corymorpha with the matching hydroid phase for one; and Coryne eximia, Turritopsis dohrnii and Turritopsis nutricula with the corresponding hydroid phases. The actinula larvae and hydroid for the pink-hearted hydroid Ectopleura crocea were identified and linked by DNA barcoding. In the order Leptothecata (thecate hydroids) medusae were identified for Clytia elsaeoswaldae, Clytia gracilis and Clytia sp. 701 AC and matched with the hydroid phases for the latter two species. Medusae were matched with the hydroid phases for two species of Obelia (including O. dichotoma) and Eucheilota bakeri. Obelia geniculata was collected as a single hydroid. DNA barcodes were obtained for hydroids of Orthopyxis everta and three other species of Orthopyxis. One member of the family Solmarisidae, representing the order Narcomedusae, and one member (Liriope tetraphylla) of the order Trachymedusae were recognized as medusae. The results show the utility of DNA barcoding for matching life-cycle stages as well as for documenting the diversity of this class of organisms.

Transcriptome Characterization of Reverse Development in Turritopsis dohrnii (Hydrozoa, Cnidaria).

Medusae of Turritopsis dohrnii undergo reverse development in response to physical damage, adverse environmental conditions, or aging. Senescent, weakened or damaged medusae transform into a cluster of poorly differentiated cells (known as the cyst stage), which metamorphose back into a preceding life cycle stage, the polyp. During the metamorphosis, cell transdifferentiation occurs. The cyst represents the intermediate stage between a reverting medusa and a healthy polyp, during which cell transdifferentiation and tissue reorganization take place. Here we characterize and compare the transcriptomes of the polyp and newborn medusa stages of T. dohrnii with that of the cyst, to identify biological networks potentially involved in the reverse development and transdifferentiation processes. The polyp, medusa and cyst of T. dohrnii were sequenced through Illumina RNA-sequencing and assembled using a de novo approach, resulting in 92,569, 74,639 and 86,373 contigs, respectively. The transcriptomes were annotated and comparative analyses among the stages identified biological networks that were significantly over-and under-expressed in the cyst as compared to the polyp and medusa stages. Biological processes that occur at the cyst stage such as telomerase activity, regulation of transposable elements and DNA repair systems, and suppression of cell signaling pathways, mitotic cell division and cellular differentiation and development may be involved in T. dohrnii's reverse development and transdifferentiation. Our results are the first attempt to understand T. dohrnii's life-cycle reversal at the genetic level, and indicate possible avenues of future research on developmental strategies, cell transdifferentiation, and aging using T. dohrnii as a non-traditional in vivo system.

Embryonic development of thecate hydrozoan Gonothyraea loveni (Allman, 1859).

Progress of Evo-Devo requires broad phylogenetic sampling providing the data for comparative analysis as well as new objects suitable for experimental investigation. Representatives of the early-branching animal phylum Cnidaria and particularly hydrozoans draw great attention due to the high diversity of embryonic and post-embryonic development and life-cycles in general. Most detailed studies on embryonic development in hydrozoans were performed on the species shedding their gametes with subsequent embryo development in the water column. Widely distributed thecate hydrozoan Gonothyraea loveni broods its embryos within reduced medusae attached to the colony until development of a free-swimming metamorphosis competent planula-larva. In the current essay we present a detailed description of G. loveni embryonic development based on in vivo observations, histology, immuno-cytochemistry, and electron microscopy. Starting from early cleavage, the embryo becomes a morula without any sign of blastocoele. Gastrulation proceeds as mixed delamination and ends with parenchymula formation. The first morphological sign of primary body axis appears only in the beginning of parenchymula-preplanula transition. In mature metamorphosis competent planula only the cells of the oral two-thirds of endoderm retain proliferative activity resulting in accumulation of great number of i-cells and nematoblasts, which can be used during metamorphosis accompanied with essential reorganization of larval tissues. G. loveni demonstrates the diversity as well as evolutionary plasticity of hydrozoans development: in brooding hydrozoans embryonic and larval development is highly embryonized in comparison with the spawning species with free-swimming embryos.

CRISPR/Cas9-mediated gene knockin in the hydroid Hydractinia symbiolongicarpus.

BACKGROUND: Hydractinia symbiolongicarpus, a colonial cnidarian, is a tractable model system for many cnidarian-specific and general biological questions. Until recently, tests of gene function in Hydractinia have relied on laborious forward genetic approaches, randomly integrated transgenes, or transient knockdown of mRNAs. RESULTS: Here, we report the use of CRISPR/Cas9 genome editing to generate targeted genomic insertions in H. symbiolonigcarpus. We used CRISPR/Cas9 to promote homologous recombination of two fluorescent reporters, eGFP and tdTomato, into the Eukaryotic elongation factor 1 alpha (Eef1a) locus. We demonstrate that the transgenes are expressed ubiquitously and are stable over two generations of breeding. We further demonstrate that CRISPR/Cas9 genome editing can be used to mark endogenous proteins with FLAG or StrepII-FLAG affinity tags to enable in vivo and ex vivo protein studies. CONCLUSIONS: This is the first account of CRISPR/Cas9 mediated knockins in Hydractinia and the first example of the germline transmission of a CRISPR/Cas9 inserted transgene in a cnidarian. The ability to precisely insert exogenous DNA into the Hydractinia genome will enable sophisticated genetic studies and further development of functional genomics tools in this understudied cnidarian model.

Identification of jellyfish neuropeptides that act directly as oocyte maturation-inducing hormones.

Oocyte meiotic maturation is crucial for sexually reproducing animals, and its core cytoplasmic regulators are highly conserved between species. By contrast, the few known maturation-inducing hormones (MIHs) that act on oocytes to initiate this process are highly variable in their molecular nature. Using the hydrozoan jellyfish species Clytia and Cladonema, which undergo oocyte maturation in response to dark-light and light-dark transitions, respectively, we deduced amidated tetrapeptide sequences from gonad transcriptome data and found that synthetic peptides could induce maturation of isolated oocytes at nanomolar concentrations. Antibody preabsorption experiments conclusively demonstrated that these W/RPRPamide-related neuropeptides account for endogenous MIH activity produced by isolated gonads. We show that the MIH peptides are synthesised by neural-type cells in the gonad, are released following dark-light/light-dark transitions, and probably act on the oocyte surface. They are produced by male as well as female jellyfish and can trigger both sperm and egg release, suggesting a role in spawning coordination. We propose an evolutionary link between hydrozoan MIHs and the neuropeptide hormones that regulate reproduction upstream of MIHs in bilaterian species.

First record of Craspedacusta sowerbii Lankester, 1880 (Hydrozoa, Limnomedusae) in a natural freshwater lagoon of Uruguay, with notes on polyp stage in captivity / Primeiro registro de Craspedacusta sowerbii Lankester, 1880 (Hydrozoa, Limnomedusae) em uma lagoa natural de água doce do Uruguai, com observações da fase de pólipo em cativeiro

Abstract The freshwater cnidarian Craspedacusta sowerbii Lankester 1880, has invaded lakes and ponds as well as artificial water bodies throughout the world. The first record in Uruguay corresponding to the jellyfish was made in 1961 in two artificial fountains, with no mention of the polyp form. Although local reports of other related polyp species have been made, information on the benthic form of C. sowerbii is lacking. Here we report the finding of live frustules, solitary individuals, medusae and colonies from a natural lagoon in August 2010, allowing us to observe the morphology and behavior of the polyp stage in captivity. In addition, molecular identification and remarks on the potencial path of introduction are presented. This is the first record in Uruguay of both polyp and medusa stages of C. sowerbii in a natural water body, Del Medio Lagoon (Dpto. de Florida), Uruguay.

Resumo O cnidário Craspedacusta sowerbii Lankester 1880 de água doce, tem invadido lagos e lagoas, bem como corpos de água artificiais em todo o mundo. O primeiro registro no Uruguai correspondente à água-viva foi feita em 1961, em duas fontes artificiais, sem mencionar a forma de pólipo. Embora existem relatórios locais de pólipos de outras espécies relacionados, informações sobre a forma bentônica de C. sowerbii ainda são escassas. Neste trabalho se relata a ocorrência de frústulas vivas, de indivíduos solitários, medusas e colônias de uma lagoa natural, em agosto de 2010, onde foi possível observar a morfologia e comportamento da fase de pólipo em cativeiro. Além disso, identificação molecular e observações sobre a potencial via de entrada da espécies são apresentados. Este é o primeiro registro no Uruguai de ambos estágios de vida de C. sowerbii em um corpo de água natural, Lagoa Del Medio (Dpto. De Florida), Uruguai.

Natural products and morphogenic activity of γ-Proteobacteria associated with the marine hydroid polyp Hydractinia echinata.

Illumina 16S rRNA gene sequencing was used to profile the associated bacterial community of the marine hydroid Hydractinia echinata, a long-standing model system in developmental biology. 56 associated bacteria were isolated and evaluated for their antimicrobial activity. Three strains were selected for further in-depth chemical analysis leading to the identification of 17 natural products. Several γ-Proteobacteria were found to induce settlement of the motile larvae, but only six isolates induced the metamorphosis to the primary polyp stage within 24h. Our study paves the way to better understand how bacterial partners contribute to protection, homeostasis and propagation of the hydroid polyp.

First record of Craspedacusta sowerbii Lankester, 1880 (Hydrozoa, Limnomedusae) in a natural freshwater lagoon of Uruguay, with notes on polyp stage in captivity.

The freshwater cnidarian Craspedacusta sowerbii Lankester 1880, has invaded lakes and ponds as well as artificial water bodies throughout the world. The first record in Uruguay corresponding to the jellyfish was made in 1961 in two artificial fountains, with no mention of the polyp form. Although local reports of other related polyp species have been made, information on the benthic form of C. sowerbii is lacking. Here we report the finding of live frustules, solitary individuals, medusae and colonies from a natural lagoon in August 2010, allowing us to observe the morphology and behavior of the polyp stage in captivity. In addition, molecular identification and remarks on the potencial path of introduction are presented. This is the first record in Uruguay of both polyp and medusa stages of C. sowerbii in a natural water body, Del Medio Lagoon (Dpto. de Florida), Uruguay.

A description of two new species of the genus Erenna (Siphonophora: Physonectae: Erennidae), with notes on recently collected specimens of other Erenna species.

Two new Erenna species, E. insidiator sp. nov. and E. sirena sp. nov., are described from specimens collected in the vicinity of Monterey Bay, California, and also, for E. sirena at the southern end of the Gulf of California, Mexico. Further information on the three extant Erenna species is given, based on specimens collected in the same areas. These have enabled, for instance, the identification of three types of tentilla on the tentacles of E. cornuta Pugh, 2001, rather than the two noted on the single previously known specimen. The genus is remarkable for the presence of bioluminescent lures on the tentilla of all five species. In E. sirena sp. nov. the tentilla are also covered by a red-fluorescent layer, which was briefly described by Haddock et al. (2005), and further details are given herein. Another extraordinary feature of the colonies E. sirena sp. nov. is that the main part of the tentacle, with its tentilla, can be extended away from the siphosomal stem on a long peduncle. This phenomenon also appears to occur in E. laciniata Pugh, 2001, and has not been observed before for other physonect species.

The polyps of Oceania armata identified by DNA barcoding (Cnidaria, Hydrozoa).

The polyps of the widely distributed medusa Oceania armata have never been found in nature and only the primary polyp is known from breeding experiments. The fully developed colony is so far unknown. This report shows that DNA sequence data of the medusa stage of O. armata permits to identify several hydroid colonies from different geographic origins as the most likely polyp stage of this medusa. These hydroids had previously been misidentified as Turritopsis species, a closely related genus which also produces medusae resembling Oceania armata. It is concluded that most Turritopsis hydroids are not reliably identifiable to species level using morphological traits only. However, DNA barcodes, particularly 16S sequences, are an excellent tool to identify the species, although we still lack information on a few nominal species and the identities of some sequence-delimited clades need to be corroborated by the addition of topotype samples.

One new genus and three new species of plumulariid hydroids (Cnidaria, Hydrozoa, Plumulariidae) from the western Pacific Ocean, with a re-examination of Plumularia insignis Allman, 1883 and related taxa.

One new genus (Schizoplumularia) and three new species (Schizoplumularia vervoorti, S. geniculata and S. elegans) of plumulariids are recognized and described from large collections of plumularioid hydroids collected in New Caledonia and vicinity during several French expeditions. During taxonomic studies of these hydroids, colonies were compared with type material of Plumularia insignis Allman, 1883 and several other similar species-group taxa. As a result, three of the latter (P. flabellum Allman, 1883, P. conjuncta Billard, 1913, and P. billardi nom. nov.) are recognized as valid in addition to P. insignis. The binomen P. billardi is a replacement name for P. insignis var. gracilis Billard, 1913. In being elevated to the rank of species in this work, it becomes an invalid junior primary homonym of several others having the same name.

Hydrozoan insights in animal development and evolution.

The fresh water polyp Hydra provides textbook experimental demonstration of positional information gradients and regeneration processes. Developmental biologists are thus familiar with Hydra, but may not appreciate that it is a relatively simple member of the Hydrozoa, a group of mostly marine cnidarians with complex and diverse life cycles, exhibiting extensive phenotypic plasticity and regenerative capabilities. Hydrozoan species offer extensive opportunities to address many developmental mechanisms relevant across the animal kingdom. Here we review recent work from non-Hydra hydrozoans - hydromedusae, hydroids and siphonophores - shedding light on mechanisms of oogenesis, embryonic patterning, allorecognition, stem cell regulation and regeneration. We also highlight potential research directions in which hydrozoan diversity can illuminate the evolution of developmental processes at micro- and macro-evolutionary time scales.