Expression profiling and cellular localization of myxozoan minicollagens during nematocyst formation and sporogenesis.

In free-living cnidarians, minicollagens are major structural components in the biogenesis of nematocysts. Recent sequence mining and proteomic analysis demonstrate that minicollagens are also expressed by myxozoans, a group of evolutionarily ancient cnidarian endoparasites. Nonetheless, the presence and abundance of nematocyst-associated genes/proteins in nematocyst morphogenesis have never been studied in Myxozoa. Here, we report the gene expression profiles of three myxozoan minicollagens, ncol-1, ncol-3, and the recently identified noncanonical ncol-5, during the intrapiscine development of Myxidium lieberkuehni, the myxozoan parasite of the northern pike, Esox lucius. Moreover, we localized the myxozoan-specific minicollagen Ncol-5 in the developing myxosporean stages by Western blotting, immunofluorescence, and immunogold electron microscopy. We found that expression of minicollagens was spatiotemporally restricted to developing nematocysts within the myxospores during sporogenesis. Intriguingly, Ncol-5 is localized in the walls of nematocysts and predominantly in nematocyst tubules. Overall, we demonstrate that despite being significantly reduced in morphology, myxozoans retain structural components associated with nematocyst development in free-living cnidarians. Furthermore, our findings have practical implications for future functional and comparative studies as minicollagens are useful markers of the developmental phase of myxozoan parasites.

Muscle systems and motility of early animals highlighted by cnidarians from the basal Cambrian.

Although fossil evidence suggests that various animal groups were able to move actively through their environment in the early stages of their evolution, virtually no direct information is available on the nature of their muscle systems. The origin of jellyfish swimming, for example, is of great interest to biologists. Exceptionally preserved muscles are described here in benthic peridermal olivooid medusozoans from the basal Cambrian of China (Kuanchuanpu Formation, ca. 535 Ma) that have direct equivalent in modern medusozoans. They consist of circular fibers distributed over the bell surface (subumbrella) and most probably have a myoepithelial origin. This is the oldest record of a muscle system in cnidarians and more generally in animals. This basic system was probably co-opted by early Cambrian jellyfish to develop capacities for jet-propelled swimming within the water column. Additional lines of fossil evidence obtained from ecdysozoans (worms and panarthropods) show that the muscle systems of early animals underwent a rapid diversification through the early Cambrian and increased their capacity to colonize a wide range of habitats both within the water column and sediment at a critical time of their evolutionary radiation.

Molecular and morphological description of Myxidium parvum (Cnidaria) from Salaria pavo (Blenniidae) in the Black Sea.

Members of the predominantly coelozioc genus Myxidum Bütschli, 1882 with more than 232 species have been reported from a wide variety of marine and freshwater fish species worldwide. In this study, 25 specimens of peacock blenny, Salaria pavo, were collected from Sinop on the Turkish Black Sea coast. The gills, fins, skin, urinary bladder, gal bladder, kidney, liver, gonads and smooth muscle tissue of the collected samples were investigated for myxosporean parasites. Myxidium parvum Yurakhno, 1991 was the only myxosporean found in the gall bladder of host fishes. Based on spore morphology, M. parvum had mostly overlapping measurement data of original description in spore length and width, polar capsule length but differed slightly in width; however, they were within the ranges previously reported from other blenniid host fish species in the Black Sea. Moreover, in this study, molecular analysis of the 18S rDNA gene of M. parvum isolates from S. pavo was done for the first time and our M. parvum genotypes appeared as sister to Myxidium incurvatum within the "Lineage II" of the marine Myxidium clade.

A comparison of the structure and function of nematocysts in free-living and parasitic cnidarians (Myxozoa).

Myxozoans are obligate parasites that have complex life cycles requiring alternate vertebrate and invertebrate hosts, with transmission via microscopic waterborne spores. Unusually for parasites, they belong to the phylum Cnidaria, alongside thousands of free-living corals, sea anemones, jellyfish and hydrozoans. Their cnidarian affinity is affirmed by genetic relatedness and the presence of nematocysts, historically called "polar capsules" in myxozoan research. Free-living cnidarians utilise this cellular weaponry for defence, predation and adhesion, whereas myxozoans use it to anchor to their hosts as the first step in infection. Despite the ~650 million years of divergence between free-living cnidarians and myxozoans, their nematocysts retain many shared morphological and molecular characters. Both are intra-cellular capsules with a single opening, and contain a coiled, evertable tubule. They are composed of unique nematocyst proteins, nematogalectin and minicollagen, and both likely contain an internal matrix of metal cations covalently bound to the anionic polymer poly-gamma glutamate. The rapid dissociation of this matrix and the resulting increase in internal osmotic potential is the driving force behind tubule elongation during discharge. In this review, we compare the structure and function of nematocysts in Myxozoa and free-living Cnidaria, incorporating recent molecular characterizations. We propose that terminology for homologous myxozoan structures be synonymized with those from other Cnidaria, hence, "polar capsule" as a taxon-specific nematocyst morphotype and "polar filament" as "tubule." Despite taxonomic divergence, genome reduction and an evolution to parasitism, myxozoans maintain nematocysts that are structurally and functionally homologous to those of their free-living cnidarian relatives.

Morphological and molecular descriptions of Sphaeromyxa sevastopoli (Cnidaria) from host fishes from Sinop on the Black Sea coast.

Members of the genus Sphaeromyxa Thélohan, 1892 have been reported from a wide variety of fish species worldwide. In the present study, specimens of rusty blenny, Parablennius sanguinolentus, collected from Sinop on the Turkish Black Sea coast were investigated for myxosporean parasites by using both conventional and molecular methods. Sphaeromyxa sevastopoli Naidenova 1970 was the only myxosporean parasite found in the gall bladder of host fishes. The morphology peculiarities of obtained S. sevastopoli spores are in good agreement with those of original description and the morphometric data overlapped in spore length and width but differed in polar capsule length and width; however, they were within the ranges previously reported from 18 host fish species. Moreover, in the present study, molecular analysis of the 18S rDNA gene of S. sevastopoli isolate in our P. sanguinolentus as well as isolates from shore rockling Gaidropsarus mediterraneus and knout goby Mesogobius batrachocephalus which were previously morphologically identified and reported by Okkay and Özer (Acta Zool Bulg 72(1):123-130, 2020) was done for the first time and our three S. sevastopoli genotypes were allocated to the "balbianii" group which is characterized by straight or slightly curved and fusiform or ovoid spores with ovoid polar capsules.

Size and density of upside-down jellyfish, Cassiopea sp., and their impact on benthic fluxes in a Caribbean lagoon.

Anthropogenic disturbances may be increasing jellyfish populations globally. Epibenthic jellyfish are ideal organisms for studying this phenomenon due to their sessile lifestyle, broad geographic distribution, and prevalence in near-shore coastal environments. There are few studies, however, that have documented epibenthic jellyfish abundance and measured their impact on ecological processes in tropical ecosystems. In this study, the density and size of the upside-down jellyfish (Cassiopea spp.) were measured in Codrington Lagoon, Barbuda. A sediment core incubation study, with and without Cassiopea, also was performed to determine their impact on benthic oxygen and nutrient fluxes. Densities of Cassiopea were 24-168 m-2, among the highest reported values in the literature. Under illuminated conditions, Cassiopea increased oxygen production >300% compared to sediment alone, and they changed sediments from net heterotrophy to net autotrophy. Cassiopea increased benthic ammonium uptake, but reduced nitrate uptake, suggesting they can significantly alter nitrogen cycling. Future studies should quantify the abundance of Cassiopea and measure their impacts on ecosystem processes, in order to further determine how anthropogenic-related changes may be altering the function of tropical coastal ecosystems.

Discovery of bilaterian-type through-guts in cloudinomorphs from the terminal Ediacaran Period.

The fossil record of the terminal Ediacaran Period is typified by the iconic index fossil Cloudina and its relatives. These tube-dwellers are presumed to be primitive metazoans, but resolving their phylogenetic identity has remained a point of contention. The root of the problem is a lack of diagnostic features; that is, phylogenetic interpretations have largely centered on the only available source of information-their external tubes. Here, using tomographic analyses of fossils from the Wood Canyon Formation (Nevada, USA), we report evidence of recognizable soft tissues within their external tubes. Although alternative interpretations are plausible, these internal cylindrical structures may be most appropriately interpreted as digestive tracts, which would be, to date, the earliest-known occurrence of such features in the fossil record. If this interpretation is correct, their nature as one-way through-guts not only provides evidence for establishing these fossils as definitive bilaterians but also has implications for the long-debated phylogenetic position of the broader cloudinomorphs.

A morphological, molecular, and histopathological redescription of Henneguya nyongensis Fomena & Bouix, 1996 (Cnidaria: Myxobolidae) infecting the gills of Peter's elephantnose fish, Gnathonemus petersii (Günther) (Osteoglossiformes: Mormyridae), imported from Nigeria.

A Henneguya sp., morphologically resembling Henneguya nyongensis Fomena & Bouix, 1996, was isolated from the gills of Peter's elephantnose fish, Gnathonemus petersii Günther, imported from Nigeria. Plasmodia were located between lamellae and within the gill epithelium, often leading to lamellar fusion. Although slightly smaller, the myxospores from these fish were morphologically consistent with H. nyongensis. In valvular view, spores are elongate, pyriform with a rounded posterior and tapering caudal processes. Myxospore bodies are 9.6-12.3 (mean 11.2) µm long and 4.0-4.7 (mean 4.3) µm wide. Polar capsules are pyriform, elongate, 4.5-5.2 (4.7) µm long and 1.3-1.6 (1.4) µm wide, with a characteristic neck-like structure at the apical end. Sequence generated for the 18S small subunit rRNA gene did not directly match any sequences available on GenBank, but demonstrated 91% nucleotide similarity to an unpublished Henneguya sp. infecting Mormyrus kannume Forsskål. Herein, the description of H. nyongensis is supplemented with new data on histopathology, molecular characterisation, and expanded host and geographical range.

On the impact of Citizen Science-derived data quality on deep learning based classification in marine images.

The evaluation of large amounts of digital image data is of growing importance for biology, including for the exploration and monitoring of marine habitats. However, only a tiny percentage of the image data collected is evaluated by marine biologists who manually interpret and annotate the image contents, which can be slow and laborious. In order to overcome the bottleneck in image annotation, two strategies are increasingly proposed: "citizen science" and "machine learning". In this study, we investigated how the combination of citizen science, to detect objects, and machine learning, to classify megafauna, could be used to automate annotation of underwater images. For this purpose, multiple large data sets of citizen science annotations with different degrees of common errors and inaccuracies observed in citizen science data were simulated by modifying "gold standard" annotations done by an experienced marine biologist. The parameters of the simulation were determined on the basis of two citizen science experiments. It allowed us to analyze the relationship between the outcome of a citizen science study and the quality of the classifications of a deep learning megafauna classifier. The results show great potential for combining citizen science with machine learning, provided that the participants are informed precisely about the annotation protocol. Inaccuracies in the position of the annotation had the most substantial influence on the classification accuracy, whereas the size of the marking and false positive detections had a smaller influence.

Towards Non-Invasive Methods to Assess Population Structure and Biomass in Vulnerable Sea Pen Fields.

Colonies of the endangered red sea pen Pennatula rubra (Cnidaria: Pennatulacea) sampled by trawling in the northwestern Mediterranean Sea were analyzed. Biometric parameters, such as total length, peduncle length, number of polyp leaves, fresh weight, and dry weight, were measured and related to each other by means of regression analysis. Ad hoc models for future inferencing of colonies size and biomass through visual techniques were individuated in order to allow a non-invasive study of the population structure and dynamics of P. rubra.

Evolution of the bilaterian mouth and anus.

It is widely held that the bilaterian tubular gut with mouth and anus evolved from a simple gut with one major gastric opening. However, there is no consensus on how this happened. Did the single gastric opening evolve into a mouth, with the anus forming elsewhere in the body (protostomy), or did it evolve into an anus, with the mouth forming elsewhere (deuterostomy), or did it evolve into both mouth and anus (amphistomy)? These questions are addressed by the comparison of developmental fates of the blastopore, the opening of the embryonic gut, in diverse animals that live today. Here we review comparative data on the identity and fate of blastoporal tissue, investigate how the formation of the through-gut relates to the major body axes, and discuss to what extent evolutionary scenarios are consistent with these data. Available evidence indicates that stem bilaterians had a slit-like gastric opening that was partially closed in subsequent evolution, leaving open the anus and most likely also the mouth, which would favour amphistomy. We discuss remaining difficulties, and outline directions for future research.

Prolific Origination of Eyes in Cnidaria with Co-option of Non-visual Opsins.

Animal eyes vary considerably in morphology and complexity and are thus ideal for understanding the evolution of complex biological traits [1]. While eyes evolved many times in bilaterian animals with elaborate nervous systems, image-forming and simpler eyes also exist in cnidarians, which are ancient non-bilaterians with neural nets and regions with condensed neurons to process information. How often eyes of varying complexity, including image-forming eyes, arose in animals with such simple neural circuitry remains obscure. Here, we produced large-scale phylogenies of Cnidaria and their photosensitive proteins and coupled them with an extensive literature search on eyes and light-sensing behavior to show that cnidarian eyes originated at least eight times, with complex, lensed-eyes having a history separate from other eye types. Compiled data show widespread light-sensing behavior in eyeless cnidarians, and comparative analyses support ancestors without eyes that already sensed light with dispersed photoreceptor cells. The history of expression of photoreceptive opsin proteins supports the inference of distinct eye origins via separate co-option of different non-visual opsin paralogs into eyes. Overall, our results show eyes evolved repeatedly from ancestral photoreceptor cells in non-bilaterian animals with simple nervous systems, co-opting existing precursors, similar to what occurred in Bilateria. Our study underscores the potential for multiple, evolutionarily distinct visual systems even in animals with simple nervous systems.

The evolution of asymmetric photosensitive structures in metazoans and the Nodal connection.

Asymmetries are observed in a great number of taxa in metazoans. More particularly, functional lateralization and neuroanatomical asymmetries within the central nervous system have been a matter of intense research for at least two hundred years. While asymmetries of some paired structures/organs (e.g. eyes, ears, kidneys, legs, arms) constitute random deviations from a pure bilateral symmetry, brain asymmetries such as those observed in the cortex and epithalamus are directional. This means that molecular and anatomical features located on one side of a given structure are observed in most individuals. For instance, in humans, the neuronal tract connecting the language areas is enlarged in the left hemisphere. When asymmetries are fixed, their molecular mechanisms can be studied using mutants displaying different phenotypes: left or right isomerism of the structure, reversed asymmetry or random asymmetry. Our understanding of asymmetry in the nervous system has been widely enriched thanks to the characterization of mutants affecting epithalamus asymmetry. Furthermore, two decades ago, pioneering studies revealed that a specific morphogen, Nodal, active only on one side of the embryo during development is an important molecule in asymmetry patterning. In this review, I have gathered important data bringing insight into the origin and evolution of epithalamus asymmetry and the role of Nodal in metazoans. After a short introduction on brain asymmetries (chapter I), I secondly focus on the molecular and anatomical characteristics of the epithalamus in vertebrates and explore some functional aspects such as its photosensitive ability related to the pineal complex (chapter II). Third, I discuss homology relationship of the parapineal organ among vertebrates (chapter III). Fourth, I discuss the possible origin of the epithalamus, presenting cells displaying photosensitive properties and/or asymmetry in the anterior part of the body in non-vertebrates (chapter IV). Finally, I report Nodal signaling expression data and functional experiments performed in different metazoan groups (chapter V).

Getting Nervous: An Evolutionary Overhaul for Communication.

The evolution of a nervous system as a control system of the body's functions is a key innovation of animals. Its fundamental units are neurons, highly specialized cells dedicated to fast cell-cell communication. Neurons pass signals to other neurons, muscle cells, or gland cells at specialized junctions, the synapses, where transmitters are released from vesicles in a Ca2+-dependent fashion to activate receptors in the membrane of the target cell. Reconstructing the origins of neuronal communication out of a more simple process remains a central challenge in biology. Recent genomic comparisons have revealed that all animals, including the nerveless poriferans and placozoans, share a basic set of genes for neuronal communication. This suggests that the first animal, the Urmetazoan, was already endowed with neurosecretory cells that probably started to connect into neuronal networks soon afterward. Here, we discuss scenarios for this pivotal transition in animal evolution.

Three Cambrian fossils assembled into an extinct body plan of cnidarian affinity.

The early Cambrian problematica Xianguangia sinica, Chengjiangopenna wangii, and Galeaplumosus abilus from the Chengjiang biota (Yunnan, China) have caused much controversy in the past and their phylogenetic placements remain unresolved. Here we show, based on exceptionally preserved material (85 new specimens plus type material), that specimens previously assigned to these three species are in fact parts of the same organism and propose that C. wangii and G. abilus are junior synonyms of X. sinica Our reconstruction of the complete animal reveals an extinct body plan that combines the characteristics of the three described species and is distinct from all known fossil and living taxa. This animal resembled a cnidarian polyp in overall morphology and having a gastric cavity partitioned by septum-like structures. However, it possessed an additional body cavity within its holdfast, an anchoring pit on the basal disk, and feather-like tentacles with densely ciliated pinnules arranged in an alternating pattern, indicating that it was a suspension feeder rather than a predatory actiniarian. Phylogenetic analyses using Bayesian inference and maximum parsimony suggest that X. sinica is a stem-group cnidarian. This relationship implies that the last common ancestor of X. sinica and crown cnidarians was probably a benthic, polypoid animal with a partitioned gastric cavity and a single mouth/anus opening. This extinct body plan suggests that feeding strategies of stem cnidarians may have been drastically different from that of their crown relatives, which are almost exclusively predators, and reveals that the morphological disparity of total-group Cnidaria is greater than previously assumed.

The nematocyst's sting is driven by the tubule moving front.

The nematocyst is the explosive injection system of the phylum Cnidaria, and is one of the fastest delivery systems found in Nature. Exploring its injection mechanism is key for understanding predator-prey interactions and protection against jellyfish stinging. Here we analyse the injection of jellyfish nematocysts and ask how the build-up of the poly-γ-glutamate (pγGlu) osmotic potential inside the nematocyst drives its discharge. To control the osmotic potential, we used a two-channel microfluidic system to direct the elongating nematocyst tubule through oil, where no osmotic potential can develop, while keeping the nematocyst capsule in water at all times. In addition, the flow inside the tubule and the pγGlu concentration profiles were calculated by applying a one-dimensional mathematical model. We found that tubule elongation through oil is orders of magnitude slower than through water and that the injection rate of the nematocyst content is reduced. These results imply that the capsule's osmotic potential is not sufficient to drive the tubule beyond the initial stage. Our proposed model shows that the tubule is pulled by the high osmotic potential that develops at the tubule moving front. This new understanding is vital for future development of nematocyst-based systems such as osmotic nanotubes and transdermal drug delivery.

Evolution of ligands, receptors and metabolizing enzymes of thyroid signaling.

Thyroid hormones (THs) play important roles in vertebrates such as the control of the metabolism, development and seasonality. Given the pleiotropic effects of thyroid disorders (developmental delay, mood disorder, tachycardia, etc), THs signaling is highly investigated, specially using mammalian models. In addition, the critical role of TH in controlling frog metamorphosis has led to the use of Xenopus as another prominent model to study THs action. Nevertheless, animals regarded as non-model species can also improve our understanding of THs signaling. For instance, studies in amphioxus highlighted the role of Triac as a bona fide thyroid hormone receptor (TR) ligand. In this review, we discuss our current understanding of the THs signaling in the different taxa forming the metazoans (multicellular animals) group. We mainly focus on three actors of the THs signaling: the ligand, the receptor and the deiodinases, enzymes playing a critical role in THs metabolism. By doing so, we also pinpoint many key questions that remain unanswered. How can THs accelerate metamorphosis in tunicates and echinoderms while their TRs have not been yet demonstrated as functional THs receptors in these species? Do THs have a biological effect in insects and cnidarians even though they do not have any TR? What is the basic function of THs in invertebrate protostomia? These questions can appear disconnected from pharmacological issues and human applications, but the investigation of THs signaling at the metazoans scale can greatly improve our understanding of this major endocrinological pathway.

Muscle and nerve net organization in stalked jellyfish (Medusozoa: Staurozoa).

Staurozoan cnidarians display an unusual combination of polyp and medusa characteristics and their morphology may be informative about the evolutionary origin of medusae. We studied neuromuscular morphology of two staurozoans, Haliclystus 'sanjuanensis' and Manania handi, using whole mount immunohistochemistry with antibodies against FMRFamide and α-tubulin to label neurons and phalloidin to label muscles. All muscles appeared to lack striations. Longitudinal interradial muscles are probable homologues of stalk muscles in scyphopolyps, but in adult staurozoans they are elaborated to inwardly flex marginal lobes of the calyx during prey capture; these muscles are pennate in M. handi. Manubrial perradial muscles, like the manubrium itself, are an innovation shared with pelagic medusae and manubrial interradial muscles are shared with scyphozoan ephyra. Marginal muscles of M. handi displayed occasional synchronous contraction reminiscent of a medusa swim pulse, but contractions were not repetitive. The nerve net in both species showed regional variation in density and orientation of neurons. Some areas labeled predominantly by α-tubulin antibodies (exumbrellar epidermis), other areas labeled exclusively by FMRFamide antibodies (dense plexus of neurites surrounding the base of secondary tentacles, neuronal concentration at the base of transformed primary tentacles; gastrodermal nerve net), but most areas showed a mix of neurons labeled by these two antibodies and frequent co-labeling of neurons. Transformed primary tentacles had a concentration of FMRFamide-immunoreactive neurons at their base that was associated with a pigment spot in M. handi; this is consistent with their homology with rhopalia of medusae, which are also derived from primary tentacles. The muscular system of these staurozoans embodies characteristics of both scyphopolyps and pelagic medusae. However, their nerve net is more polyp-like, although marginal concentrations of the net associated with primary and secondary tentacles may facilitate the richer behavioral repertoire of staurozoans relative to polyps of other medusozoans. J. Morphol. 278:29-49, 2017. ©© 2016 Wiley Periodicals,Inc.

Evolution of eumetazoan nervous systems: insights from cnidarians.

Cnidarians, the sister group to bilaterians, have a simple diffuse nervous system. This morphological simplicity and their phylogenetic position make them a crucial group in the study of the evolution of the nervous system. The development of their nervous systems is of particular interest, as by uncovering the genetic programme that underlies it, and comparing it with the bilaterian developmental programme, it is possible to make assumptions about the genes and processes involved in the development of ancestral nervous systems. Recent advances in sequencing methods, genetic interference techniques and transgenic technology have enabled us to get a first glimpse into the molecular network underlying the development of a cnidarian nervous system-in particular the nervous system of the anthozoan Nematostella vectensis. It appears that much of the genetic network of the nervous system development is partly conserved between cnidarians and bilaterians, with Wnt and bone morphogenetic protein (BMP) signalling, and Sox genes playing a crucial part in the differentiation of neurons. However, cnidarians possess some specific characteristics, and further studies are necessary to elucidate the full regulatory network. The work on cnidarian neurogenesis further accentuates the need to study non-model organisms in order to gain insights into processes that shaped present-day lineages during the course of evolution.