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Nervous systems of bilaterian animals generally consist of two cell types: neurons and glial cells. Despite accumulating data about the many important functions glial cells serve in bilaterian nervous systems, the evolutionary origin of this abundant cell type remains unclear. Current hypotheses regarding glial evolution are mostly based on data from model bilaterians. Non-bilaterian animals have been largely overlooked in glial studies and have been subjected only to morphological analysis. Here, we provide a comprehensive overview of conservation of the bilateral gliogenic genetic repertoire of non-bilaterian phyla (Cnidaria, Placozoa, Ctenophora, and Porifera). We overview molecular and functional features of bilaterian glial cell types and discuss their possible evolutionary history. We then examine which glial features are present in non-bilaterians. Of these, cnidarians show the highest degree of gliogenic program conservation and may therefore be crucial to answer questions about glial evolution., (© 2024. The Author(s).)

Medusozoans, the Cnidarian subphylum, have multiple life stages including sessile polyps and free-swimming medusae or jellyfish, which are typically bell-shaped gelatinous zooplanktons that exhibit diverse morphologies. Despite having a relatively complex body structure with well-developed muscles and nervous systems, the adult medusa stage maintains a high regenerative ability that enables organ regeneration as well as whole body reconstitution from the part of the body. This remarkable regeneration potential of jellyfish has long been acknowledged in different species; however, recent studies have begun dissecting the exact processes underpinning regeneration events. In this article, we introduce the current understanding of regeneration mechanisms in medusae, particularly focusing on cellular behaviors during regeneration such as wound healing, blastema formation by stem/progenitor cells or cell fate plasticity, and the organism-level patterning that restores radial symmetry. We also discuss putative molecular mechanisms involved in regeneration processes and introduce a variety of novel model jellyfish species in the effort to understand common principles and diverse mechanisms underlying the regeneration of complex organs and the entire body.

Disruption of the continuous cutaneous membrane in the integumentary system is considered a health problem of high cost for any nation. Several attempts have been made for developing skin substitutes in order to restore injured tissue including autologous implants and the use of scaffolds based on synthetic and natural materials. Current biomaterials used for skin tissue repair include several scaffold matrices types, synthetic or natural, absorbable, degradable or non-degradable polymers, porous or dense scaffolds, and cells capsulated in hydrogels or spheroids systems so forth. These materials have advantages and disadvantages and its use will depend on the desired application. Recently, marine organisms such as jellyfish have attracted renewed interest, because both its composition and structure resemble the architecture of human dermic tissue. In this context, the present study aims to generate scaffolds from Cassiopea andromeda (C. andromeda), with application in skin tissue engineering, using a decellularization process. The obtained scaffold was studied by infrared spectroscopy (FT-IR), thermogravimetric analysis (TGA), differential scanning calorimetry analysis (DSC), and scanning electron microscopy (SEM). Crystal violet staining and DNA quantification assessed decellularization effectiveness while the biocompatibility of scaffold was determined with human dermic fibroblasts. Results indicated that the decellularization process reduce native cell population leading to 70% reduction in DNA content. In addition, SEM showed that the macro and microstructure of the collagen I-based scaffold were preserved allowing good adhesion and proliferation of human dermic fibroblasts. The C. andromeda scaffold mimics human skin and therefore represents great potential for skin tissue engineering., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

In Cnidaria, modes of gastrulation to produce the two body layers vary greatly between species. In the hydrozoan species Clytia hemisphaerica gastrulation involves unipolar ingression of presumptive endoderm cells from an oral domain of the blastula, followed by migration of these cells to fill the blastocoel with concomitant narrowing of the gastrula and elongation along the oral-aboral axis. We developed a 2D computational boundary model capable of simulating the morphogenetic changes during embryonic development from early blastula stage to the end of gastrulation. Cells are modeled as polygons with elastic membranes and cytoplasm, colliding and adhering to other cells, and capable of forming filopodia. With this model we could simulate compaction of the embryo preceding gastrulation, bottle cell formation, ingression, and intercalation between cells of the ingressing presumptive endoderm. We show that embryo elongation is dependent on the number of endodermal cells, low endodermal cell-cell adhesion, and planar cell polarity (PCP). When the strength of PCP is reduced in our model, resultant embryo morphologies closely resemble those reported previously following morpholino-mediated knockdown of the core PCP proteins Strabismus and Frizzled. Based on our results, we postulate that cellular processes of apical constriction, compaction, ingression, and then reduced cell-cell adhesion and mediolateral intercalation in the presumptive endoderm, are required and when combined, sufficient for Clytia gastrulation., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

Epithelial folding (EF) is a fundamental morphogenetic process that can be observed in the development of many organisms ranging from metazoans to green algae. Being early branching metazoans, cnidarians represent the best models to study evolutionarily conserved morphogenetic processes, including EF. Hydrozoa is the most evolutionary advanced group of the phylum Cnidaria. All colonial hydrozoans grow continuously, changing the shape of their colonies and spreading over the substrate with the help of elongating stolons. Owing to high diversity of colony architecture, they are ideal objects for comparative and evolutionary morphology. In the hydrozoan Dynamena pumila, the growth of the colony proceeds via a variety of morphogenetic processes. Our work is focused on the formation of the anchoring disc of the stolon, which is accompanied by inward-folding morphogenesis of the ectodermal layer. Successive stages of anchoring disc development were described with light, confocal transmission electron microscopy. We have shown that EF in Dynamena is associated with accumulation of F-actin in the constricting apical domains of forming bottle cells located at the bottom of the emerging fold. In addition, the nuclei of these cells are displaced to the basal domains. Taken together, these features may indicate that EF in Dynamena proceeds as an active invagination, although this process has never been described in the development of hydrozoans. Apparently, development of the anchoring disc can be viewed as a reliable and versatile model system for studying the cell-shape-change-driven epithelial sheet morphogenesis, which can be easily observed and analysed., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Polypodium hydriforme, the only species in Polypodiozoa, which is currently considered a class of Cnidaria, and likely a sister group to Medusozoa (together with Myxozoa), is a cnidarian adapted to intracellular parasitism inside sturgeon oocytes. Free-living P. hydriforme lives on river bottoms; it walks on supporting tentacles and uses sensory tentacles to capture food and bring it to the mouth. The nervous system of free-living P. hydriforme was studied by confocal microscopy and immunohistochemistry using antibodies to FMRF-amide and α-tubulin combined with phalloidin-staining of F-actin fibres. A sensory FMRF-amide immunoreactive (IR) nerve net and an α-tubulin IR nerve net have been identified. The FMRF-amide IR nerve net underlies the epidermis along the tentacles and around the mouth; it consists of neurites emanating from epidermal sensory cells and basiepidermal ganglion cells, and it connects with cnidocytes. A deeper-lying α-tubulin IR nerve net occurs only in tentacles and looks like chains of different-sized beads crossing the mesoglea and entwining muscles. Anti-α-tubulin staining also reveals microtubules in muscle cells following the longitudinal muscle fibres or the thin circular F-actin fibres of the tentacles. Cnidocytes in the tentacles are embedded in a regular hexagonal non-neural network formed by the tubulin IR cytoskeleton of epidermal cells. Cnidocils of the cnidocytes around the mouth and in walking tentacles are identical, but those in sensory tentacles differ in length and width. The possible homology of the tubulin IR nerve net with motor nerve nets of cnidarians is discussed. The absence of a classic nerve ring around the mouth and the lack of specialised sense organs are considered to be plesiomorphic characters for Cnidaria., (Copyright © 2015 Elsevier GmbH. All rights reserved.)

The data revealed by comparative embryology of the basal (diploblastic) metazoans is traditionally considered a valuable potential source of information on the origin and early evolution of the animal kingdom and its major clades. Special attention is paid to the fundamental morphogenetic process of gastrulation during which the cells of the early embryo differentiate into the germ layers and the primary body plan is formed. Comparative analysis of gastrulation in different cnidarian taxa reveals high level of intergroup, intragroup, and individual variation. With few exceptions, there is no robust correlation between the type of gastrulation and the taxon. Current data do not support the idea that morphogenetic processes underlying cnidarian gastrulation can be divided into several distinct types. Rather, there is a continuum of equifinal ontogenetic trajectories. In cnidarians, the mode of gastrulation apparently depends less on the macroevolutionary history of the species than on various evolutionary plastic features, such as the oocyte size, the amount of yolk, the number of cells at the blastula (or morula) stage, the presence of phototrophic symbionts, or the ecology of the larva. Thus, in cnidarians, morphogenetic basis of gastrulation contains only a very weak phylogenetic signal and can have only limited application in phylogenetic reconstructions. On the other hand, comparative studies of the ontogeny of the basal metazoans shed light on the general rules of the evolution of morphogenetic processes that is crucial for understanding the early history of the animal kingdom.

In studies of both the establishment and breakdown of cnidarian-dinoflagellate symbiosis, it is often necessary to determine the number of Symbiodinium cells relative to the quantity of host tissue. Ideally, the methods used should be rapid, precise, and accurate. In this study, we systematically evaluated methods for sample preparation and storage and the counting of algal cells using the hemocytometer, a custom image-analysis program for automated counting of the fluorescent algal cells, the Coulter Counter, or the Millipore Guava flow-cytometer. We found that although other methods may have value in particular applications, for most purposes, the Guava flow cytometer provided by far the best combination of precision, accuracy, and efficient use of investigator time (due to the instrument's automated sample handling), while also allowing counts of algal numbers over a wide range and in small volumes of tissue homogenate. We also found that either of two assays of total homogenate protein provided a precise and seemingly accurate basis for normalization of algal counts to the total amount of holobiont tissue.

Cnidarians possess remarkable powers of regeneration, but the cellular and molecular mechanisms underlying this capability are unclear. Studying the hydrozoan Hydractinia echinata we show that a burst of stem cell proliferation occurs following decapitation, forming a blastema at the oral pole within 24 hr. This process is necessary for head regeneration. Knocking down Piwi1, Vasa, Pl10 or Ncol1 expressed by blastema cells inhibited regeneration but not blastema formation. EdU pulse-chase experiments and in vivo tracking of individual transgenic Piwi1(+) stem cells showed that the cellular source for blastema formation is migration of stem cells from a remote area. Surprisingly, no blastema developed at the aboral pole after stolon removal. Instead, polyps transformed into stolons and then budded polyps. Hence, distinct mechanisms act to regenerate different body parts in Hydractinia. This model, where stem cell behavior can be monitored in vivo at single cell resolution, offers new insights for regenerative biology.

Understanding how new cell types arise is critical for understanding the evolution of organismal complexity. Questions of this nature, however, can be difficult to answer due to the challenge associated with defining the identity of a truly novel cell. Cnidarians (anemones, jellies, and their allies) provide a unique opportunity to investigate the molecular regulation and development of cell-novelty because they possess a cell that is unique to the cnidarian lineage and that also has a very well-characterized phenotype: the cnidocyte (stinging cell). Because cnidocytes are thought to differentiate from the cell lineage that also gives rise to neurons, cnidocytes can be expected to express many of the same genes expressed in their neural "sister" cells. Conversely, only cnidocytes posses a cnidocyst (the explosive organelle that gives cnidocytes their sting); therefore, those genes or gene-regulatory relationships required for the development of the cnidocyst can be expected to be expressed uniquely (or in unique combination) in cnidocytes. This system provides an important opportunity to: (1) construct the gene-regulatory network (GRN) underlying the differentiation of cnidocytes, (2) assess the relative contributions of both conserved and derived genes in the cnidocyte GRN, and (3) test hypotheses about the role of novel regulatory relationships in the generation of novel cell types. In this review, we summarize common challenges to studying the evolution of novelty, introduce the utility of cnidocyte differentiation in the model cnidarian, Nematostella vectensis, as a means of overcoming these challenges, and describe an experimental approach that leverages comparative tissue-specific transcriptomics to generate hypotheses about the GRNs underlying the acquisition of the cnidocyte identity., (© The Author 2014. Published by Oxford University Press on behalf of the Society for Integrative and Comparative Biology. All rights reserved. For permissions please email: journals.permissions@oup.com.)

Background: Wound healing is the first stage of a series of cellular events that are necessary to initiate a regenerative response. Defective wound healing can block regeneration even in animals with a high regenerative capacity. Understanding how signals generated during wound healing promote regeneration of lost structures is highly important, considering that virtually all animals have the ability to heal but many lack the ability to regenerate missing structures. Cnidarians are the phylogenetic sister taxa to bilaterians and are highly regenerative animals. To gain a greater understanding of how early animals generate a regenerative response, we examined the cellular and molecular components involved during wound healing in the anthozoan cnidarian Nematostella vectensis., Results: Pharmacological inhibition of extracellular signal-regulated kinases (ERK) signaling blocks regeneration and wound healing in Nematostella. We characterized early and late wound healing events through genome-wide microarray analysis, quantitative PCR, and in situ hybridization to identify potential wound healing targets. We identified a number of genes directly related to the wound healing response in other animals (metalloproteinases, growth factors, transcription factors) and suggest that glycoproteins (mucins and uromodulin) play a key role in early wound healing events. This study also identified a novel cnidarian-specific gene, for a thiamine biosynthesis enzyme (vitamin B synthesis), that may have been incorporated into the genome by lateral gene transfer from bacteria and now functions during wound healing. Lastly, we suggest that ERK signaling is a shared element of the early wound response for animals with a high regenerative capacity., Conclusions: This research describes the temporal events involved during Nematostella wound healing, and provides a foundation for comparative analysis with other regenerative and non-regenerative species. We have shown that the same genes that heal puncture wounds are also activated after oral-aboral bisection, indicating a clear link with the initiation of regenerative healing. This study demonstrates the strength of using a forward approach (microarray) to characterize a developmental phenomenon (wound healing) at a phylogenetically important crossroad of animal evolution (cnidarian-bilaterian ancestor). Accumulation of data on the early wound healing events across numerous systems may provide clues as to why some animals have limited regenerative abilities.

Cell-extracellular matrix (ECM) and cell-cell adhesion systems are fundamental to the multicellularity of metazoans. Members of phylum Cnidaria were classified historically by their radial symmetry as an outgroup to bilaterian animals. Experimental study of Hydra and jellyfish has fascinated zoologists for many years. Laboratory studies, based on dissection, biochemical isolations, or perturbations of the living organism, have identified the ECM layer of cnidarians (mesoglea) and its components as important determinants of stem cell properties, cell migration and differentiation, tissue morphogenesis, repair, and regeneration. Studies of the ultrastructure and functions of intercellular gap and septate junctions identified parallel roles for these structures in intercellular communication and morphogenesis. More recently, the sequenced genomes of sea anemone Nematostella vectensis, Hydra magnipapillata, and coral Acropora digitifera have opened up a new frame of reference for analyzing the cell-ECM and cell-cell adhesion molecules of cnidarians and examining their conservation with bilaterians. This chapter integrates a review of literature on the structure and functions of cell-ECM and cell-cell adhesion systems in cnidarians with current analyses of genome-encoded repertoires of adhesion molecules. The postgenomic perspective provides a fresh view on fundamental similarities between cnidarian and bilaterian animals and is impelling wider adoption of species from phylum Cnidaria as model organisms., (© 2014 Elsevier Inc. All rights reserved.)

Coral bleaching occurs when there is a breakdown of the symbiosis between cnidarian hosts and resident Symbiodinium spp. Multiple mechanisms for the bleaching process have been identified, including apoptosis and autophagy, and most previous work has focused on the Symbiodinium cell as the initiator of the bleaching cascade. In this work we show that it is possible for host cells to initiate apoptosis that can contribute to death of the Symbiodinium cell. First we found that colchicine, which results in apoptosis in other animals, causes cell death in the model anemone Aiptasia sp. but not in cultured Symbiodinium CCMP-830 cells or in cells freshly isolated from host Aiptasia (at least within the time frame of our study). In contrast, when symbiotic Aiptasia were incubated in colchicine, cell death in the resident Symbiodinium cells was observed, suggesting a host effect on symbiont mortality. Using live-cell confocal imaging of macerated symbiotic host cell isolates, we identified a pattern where the initiation of host cell death was followed by mortality of the resident Symbiodinium cells. This same pattern was observed in symbiotic host cells that were subjected to temperature stress. This research suggests that mortality of symbionts during temperature-induced bleaching can be initiated in part by host cell apoptosis.

The environmental contamination caused by heavy metals raises the question of their effect on biological systems. Among bio-indicators useful to monitor the toxicological effects of these chemicals, Cnidarians offer a unique model. Cnidarians possess highly specialized stinging cells, termed nematocytes, which respond to hyposmotic solution with well established homeostatic parameters as an acute osmotic phase (OP), leading to cell swelling, and then a slower regulatory volume decrease (RVD) phase, causing cell shrinkage. Here we report the effect of 65% artificial sea water (ASW) containing heavy metals, such as Cd, La, Co, Cu and Zn (concentrations comprised between 100 and 0.1 μM) on both OP and RVD in nematocytes isolated from the jellyfish Pelagia noctiluca by 605 mM NaSCN plus 0.01 mM Ca(2+). The exposure of the cells to Co and La inhibited RVD but not OP. However, Cu, Cd and Zn prevented the OP in a dose-dependent manner and, hence, also the detection of RVD. These results suggest that, in isolated nematocytes, heavy metal pollutants impair RVD either directly or indirectly through interference with the OP, thus negating RVD. Although further studies need to clarify the exact mechanisms whereby heavy metals exert their toxicity, it is evident that nematocytes of Cnidarians could serve as a model for ecotoxicological investigations., (Copyright © 2013 Elsevier Inc. All rights reserved.)

The first animals arose more than six hundred million years ago, yet they left little impression in the fossil record. Nonetheless, the cell biology and genome composition of the first animal, the Urmetazoan, can be reconstructed through the study of phylogenetically relevant living organisms. Comparisons among animals and their unicellular and colonial relatives reveal that the Urmetazoan likely possessed a layer of epithelium-like collar cells, preyed on bacteria, reproduced by sperm and egg, and developed through cell division, cell differentiation, and invagination. Although many genes involved in development, body patterning, immunity, and cell-type specification evolved in the animal stem lineage or after animal origins, several gene families critical for cell adhesion, signaling, and gene regulation predate the origin of animals. The ancestral functions of these and other genes may eventually be revealed through studies of gene and genome function in early-branching animals and their closest non-animal relatives.

The sequestration of nematocysts (a special group of cnidocysts) from cnidarian prey with subsequent use in defence is described for few metazoan phyla. Members of the taxon Aeolidoidea (Nudibranchia, Gastropoda) are well-known for this. Questions regarding the reasons some nematocysts do not discharge when the gastropod feeds and how these same nematocysts can be transported along the digestive tract into specialized morphological structures called cnidosacs, remain unanswered. Within the cnidosac, nematocysts are incorporated in cells and finally be used for defence against predators. The most plausible explanation for this phenomenon suggests there are immature and therefore non-functional nematocysts in the food. A recent study by Berking and Herrmann (2005) on cnidarians suggested that the nematocysts mature by acidification via proton transfer into the nematocyst capsule. According to this hypothesis only immature nematocysts are transported into the cnidosac where they are then made functional through an accumulation of protons. In this study we present a fluorescence staining method that tests the hypothesis by Berking and Herrmann (2005) and detects changes in the pH values of incorporated nematocysts, interpreted as changes in maturation stages. This marker, the fluorescent dye Ageladine A, stains nematocyst capsules according to their pH values. With Ageladine A we were able to show that kleptocnides indeed change their pH value after incorporation into the aeolidoidean cnidosac., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

In both plants and animals, regeneration requires the activation of stem cells. This is possibly related to the origin and requirements of multicellularity. Although long diverged from a common ancestry, plant and animal models such as Arabidopsis, Drosophila and mouse share considerable similarities in stem cell regulation. This includes stem cell niche organisation, epigenetic modification of DNA and histones, and the role of small RNA machinery in differentiation and pluripotency states. Dysregulation of any of these can lead to premature ageing, patterning and specification defects, as well as cancers. Moreover, emerging basal animal and plant systems are beginning to provide important clues concerning the diversity and evolutionary history of stem cell regulatory mechanisms in eukaryotes. This review provides a comparative framework, highlighting both the commonalities and differences among groups, which should promote the intelligent design of artificial stem cell systems, and thereby fuel the field of biomaterials science., (Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Cnidae are complex intracellular capsules made by all cnidarians. The most diverse of these capsules are nematocysts, which are made by all members of the phylum; spirocysts and ptychocysts are made only by members of some lineages, and they show less functional and structural diversity. In nematocysts, the apex has been shown to be either a hinged cap (operculum) or three flaps that flex outward during discharge. The operculum is known only from medusozoan nematocysts; flaps are known only from nematocysts of members of the hexacorallian order Actiniaria, although they have been inferred to be characteristic of Anthozoa, the group to which Actiniaria belongs. Using scanning and transmission electron microscopy, we discover a third apical morphology in nematocysts, an apical cap, which we find in all nonactiniarian anthozoans examined. This apical cap is identical structurally to the apical cap of spirocysts, and it resembles the apical structure of ptychocysts, whose apex is documented here for the first time. Additionally, a full survey of nematocysts from all body structures of two actiniarians demonstrates that a particular type of nematocyst, the microbasic p-mastigophore of the mesenterial filaments, does not have apical flaps. The observed variation does not correspond to conventional categorization of capsule morphology and raises questions about the function and structure of capsules across Cnidaria. Despite some ambiguity in optimization of ancestral states across cnidae, we determine that the apical cap is the plesiomorphic structure for anthozoan cnidae and that apical flaps are a synapomorphy of Actiniaria. At present, the operculum is interpreted as a synapomorphy for Medusozoa, but either it or an apical cap is the ancestral state for nematocysts., (Copyright © 2011 Wiley Periodicals, Inc.)

The symbiotic relationship between cnidarians and their dinoflagellate symbionts, Symbiodinium spp, which underpins the formation of tropical coral reefs, can be destabilized by rapid changes to environmental conditions. Although some studies have concluded that a breakdown in the symbiosis begins with increased reactive oxygen species (ROS) generation within the symbiont due to a decoupling of photosynthesis, others have reported the release of viable symbionts via a variety of host cell derived mechanisms. We explored an alternative model focused upon changes in host cnidarian mitochondrial integrity in response to thermal stress. Mitochondria are often likened to being batteries of the cell, providing energy in the form of ATP, and controlling cellular pathway activation and ROS generation. The overall morphology of host mitochondria was compared to that of associated symbionts under an experimental thermal stress using confocal and electron microscopy. The results demonstrate that hyperthermic stress induces the degradation of cnidarian host mitochondria that is independent of symbiont cellular deterioration. The potential sites of host mitochondrial disruption were also assessed by measuring changes in the expression of genes associated with electron transport and ATP synthesis using quantitative RT-PCR. The primary site of degradation appeared to be downstream of complex III of the electron transport chain with a significant reduction in host cytochrome c and ATP synthase expression. The consequences of reduced expression could limit the capacity of the host to mitigate ROS generation and maintain both organelle integrity and cellular energy supplies. The disruption of host mitochondria, cellular homeostasis, and subsequent cell death irrespective of symbiont integrity highlights the importance of the host response to thermal stress and in symbiosis dysfunction that has substantial implications for understanding how coral reefs will survive in the face of climate change.

Cnidarian taxa, currently of the most morphologically simplest extant metazoans, exhibit many salient properties of innate immunity that are shared by most Animalia. One hallmark constituent of immunity exhibit by most cnidarians is histocompatibility, marked by wide spectrum of allogeneic and xenogeneic effector arms, progressing into tissue fusions or inflammatory rejections. Scientific propensity on cnidarians immunity, while discussing historecognition as the ground for immunity in these organisms, concentrates on host-parasitic and disease oriented studies, or focuses on genome approaches that search for gene homologies with the vertebrates. Above tendency for mixing up between historecognition and host-parasitic/disease, highlights a serious obstacle for the progress in our understanding of cnidarian immunobiology. Here I critically overview four 'forgotten' cnidarian immune features, namely, specificity, immunological memory, allogeneic maturation and natural chimerism, presenting insights into perspectives that are prerequisite for any discussion on cnidarian evolution. It is evident that cnidarian historecognition embraces elements that the traditional field of vertebrate immunology has never encountered (i.e., variety of cytotoxic outcomes, different types of effector mechanisms, chimerism, etc.). Also, cnidarian immune features dictating that different individuals within the same species seem to respond differently to the same immunological challenge, is far from that recorded in the vertebrates' adaptive immunity. While above features may be connected to host-parasitic and disease phenomena and effector arms, they clearly attest to their unique critical roles in shaping cnidarians historecognition, calling for improved distinction between historecognition and host-response/ disease disciplines. The research on cnidarians immunity still suffers from the lack of accepted synthesis of what historecognition is or does. Mounting of an immune response against conspecifics or xenogeneic organisms should therefore be clearly demarcated from other paths of immunity, till cnidarian innate immunity as a whole is expounded.

Transitory fusion is an allorecognition phenotype displayed by the colonial hydroid Hydractinia symbiolongicarpus when interacting colonies share some, but not all, loci within the allorecognition gene complex (ARC). The phenotype is characterized by an initial fusion followed by subsequent cell death resulting in separation of the two incompatible colonies. We here characterize this cell death process using scanning electron microscopy (SEM), transmission electron microscopy (TEM), and continuous in vivo digital microscopy. These techniques reveal widespread autophagy and subsequent necrosis in both colony and grafted polyp assays. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assays and ultrastructural observations revealed no evidence of apoptosis. Pharmacological inhibition of autophagy using 3-methyladenine (3-MA) completely suppressed transitory fusion in vivo in colony assays. Rapamycin did not have a significant effect in the same assays. These results establish the hydroid allorecognition system as a novel model for the study of cell death.

The evolutionary origin of stem cell pluripotency is an unresolved question. In mammals, pluripotency is limited to early embryos and is induced and maintained by a small number of key transcription factors, of which the POU domain protein Oct4 is considered central. Clonal invertebrates, by contrast, possess pluripotent stem cells throughout their life, but the molecular mechanisms that control their pluripotency are poorly defined. To address this problem, we analyzed the expression pattern and function of Polynem (Pln), a POU domain gene from the marine cnidarian Hydractinia echinata. We show that Pln is expressed in the embryo and adult stem cells of the animal and that ectopic expression in epithelial cells induces stem cell neoplasms and loss of epithelial tissue. Neoplasm cells downregulated the transgene but expressed the endogenous Pln gene and also Nanos, Vasa, Piwi and Myc, which are all known cnidarian stem cell markers. Retinoic acid treatment caused downregulation of Pln and the differentiation of neoplasm cells to neurosensory and epithelial cells. Pln downregulation by RNAi led to differentiation. Collectively, our results suggest an ancient role of POU proteins as key regulators of animal stem cells.

We conducted gross and microscopic characterizations of lesions in Cnidaria from Palmyra Atoll, Central Pacific. We found growth anomalies (GA) to be the most commonly encountered lesion. Cases of discoloration and tissue loss were rare. GAs had a focal or multi-focal distribution and were predominantly nodular, exophytic, and umbonate. In scleractinians, the majority of GAs manifested as hyperplasia of the basal body wall (52% of cases), with an associated absence or reduction of polyp structure (mesenteries and filaments, actinopharynx and tentacles), and depletion of zooxanthellae in the gastrodermis of the upper body wall. In the soft corals Sinularia sp. and Lobophytum sp., GAs exclusively manifested as prominent hyperplasia of the coenenchyme with an increased density of solenia. In contrast to scleractinians, soft coral GAs displayed an inflammatory and necrotizing component with marked edema of the mesoglea, accompanied by infiltrates of variably-sized granular amoebocytes. Fungi, algae, sponges, and Crustacea were present in some scleractinian GAs, but absent in soft coral GAs. Fragmentation of tissues was a common finding in Acropora acuminata and Montipora cf. dilatata colonies with tissue loss, although no obvious causative agents were seen. Discoloration in the zoanthid, Palythoa tuberculosa, was found to be the result of necrosis, while in Lobophytum sp. discoloration was the result of zooxanthellar depletion (bleaching). Soft corals with discoloration or tissue loss showed a marked inflammatory response, however no obvious causative organisms were seen. Lesions that appeared similar at the gross level were revealed to be distinct by microscopy, emphasizing the importance of histopathology., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Zoanthids comprise an order of benthic, generally colonial cnidarians, which can usually be distinguished from other hexacorallians by embedded sand and detritus in their mesoglea to help strengthen their structure. These animals are becoming increasingly important research subjects in biochemistry and other research fields. Their inclusion of both calcium and silica results in the need for both decalcification and desilification for internal morphological examinations. Since the methodology of hydrofluoric acid (HF) desilification has rarely been documented in zoanthids, histological surveys for zoanthid taxonomy have often been abandoned and their taxonomy is often problematic. Recent investigations utilizing molecular methods have brought a clearer understanding of zoanthid diversity, but standardization of HF treatments are still needed to provide a link between molecular and more traditional techniques, and to properly examine specimens for which molecular methods may not be an option (e.g., formalin-preserved specimens, etc.). Here, we use both "straight" HF and, for the first time with zoanthids, buffered HF (BHF) treatments at different treatment lengths (1-48 h) on polyps from three different species of zoanthids for histological examination. Section conditions were judged based on the presence/absence of embedded detritus, drag marks, and tissue condition. Results show that the BHF treatment resulted in slightly better tissue conditions for all specimens, and suggest that desilification works well regardless of treatment time for species with smaller (polyp diameter <0.5 cm), less heavily encrusted polyps. Desilification of heavily encrusted Palythoa mutuki polyps were still problematic, with at least 24 h treatment needed. To aid future research, we provide guidelines for HF treatments of zoanthid specimens.

Sessile organisms may experience chronic exposure to copper that is released into the marine environment from antifoulants and stormwater runoff. We have identified the site of damage caused by copper to the symbiotic cnidarian, Zoanthus robustus (Anthozoa, Hexacorallia). External changes to the zoanthids were apparent when compared with controls. The normally flexible bodies contracted and became rigid. Histological examination of the zoanthid tissue revealed that copper had caused sub-cellular changes to proteins within the extracellular matrix (ECM) of the tubular body. Collagen in the ECM and the internal septa increased in thickness to five and seven times that of controls respectively. The epithelium, which stained for elastin, was also twice as thick and tough to cut, but exposure to copper did not change the total amount of desmosine which is found only in elastin. We conclude that copper stimulated collagen synthesis in the ECM and also caused cross-linking of existing proteins. However, there was no expulsion of the symbiotic algae (Symbiodinium sp.) and no effect on algal pigments or respiration (44, 66 and 110 microg Cu L(-1)). A decrease in net photosynthesis was observed only at the highest copper concentration (156 microg Cu L(-1)). These results show that cnidarians may be more susceptible to damage by copper than their symbiotic algae., (Copyright (c) 2010 Elsevier Inc. All rights reserved.)

Because cnidocytes are exceedingly complex cells which can only be used once, their discharge is highly regulated by way of a variety of chemosensory, mechanosensory and endogenous pathways. The integration of these various inputs ultimately results in exocytosis and then discharge of the cnidocyte's diagnostic organelle, the cnidocyst. Here we review what is known about the sensory pathways that regulate cnidocytes, the electrical events that manifest in cnidocytes following sensory stimulation and the ionic mechanisms that underlie discharge.

The cnidocyst is the defining organelle of the cnidarians, used for capture of prey and defense. It consists of a cylindrical capsule, which releases a long tubule upon triggering. Cnidocysts develop inside a giant post-Golgi vesicle by a sequential accumulation of proteins from the Golgi apparatus. Traditionally three types of cnidocysts are distinguished: nematocysts, spirocysts, and ptychocysts. Here we focus on nematocysts, the prototypic cnidocyst and by far most diverse group of cnidocysts in this phylum. The mature nematocyst capsule comprises a collagenous polymer with remarkable biophysical properties, able to withstand an osmotic pressure of 150 bar. Release of the capsule and discharge is probably initiated by classical exocytosis. High-speed studies revealed the kinetics of discharge to be as short as 700 ns, generating an acceleration of 5,400,000 x g and a pressure of 7.7 GPa at the site of impact of the spines onto the prey. Thus nematocysts comprise a powerful molecular spring mechanism releasing energy stored in the wall polymer in the nanosecond time range. During the last few years, genomic, biochemical and structural studies have helped to unravel the molecular composition of the nematocyst supra-structure. Here we summarize these findings and present an integrative view of mechanical and molecular aspects that have shaped the nematocyst during evolution.

Although toxic, physically destructive, and produced solely by cnidarians, nematocysts are acquired, stored, and used by some predators of cnidarians. Despite knowledge of this phenomenon for well over a century, little empirical evidence details the mechanisms of how (and even why) these organisms use organelles of cnidarians. However, in the past 20 years a number of published experimental investigations address two of the fundamental questions of nematocyst acquisition and use by cnidarian predators: (1) how are cnidarian predators protected from nematocyst discharge during feeding; and (2) how are the nematocysts used by the predator?

The kinase Mos, which activates intracellularly the MAP kinase pathway, is a key regulator of animal oocyte meiotic maturation. In vertebrate and echinoderm models, Mos RNA translation upon oocyte hormonal stimulation mediates "cytostatic" arrest of the egg after meiosis, as well as diverse earlier events [1-5]. Our phylogenetic survey has revealed that MOS genes are conserved in cnidarians and ctenophores, but not found outside the metazoa or in sponges. We demonstrated MAP kinase-mediated cytostatic activity for Mos orthologs from Pleurobrachia (ctenophore) and Clytia (cnidarian) by RNA injection into Xenopus blastomeres. Analyses of endogenous Mos in Clytia with morpholino antisense oligonucleotides and pharmacological inhibition demonstrated that Mos/MAP kinase function in postmeiotic arrest is conserved. They also revealed additional roles in spindle formation and positioning, strongly reminiscent of observations in starfish, mouse, and Xenopus. Unusually, cnidarians were found to possess multiple Mos paralogs. In Clytia, one of two maternally expressed paralogs accounted for the majority MAP kinase activation during maturation, whereas the other may be subject to differential translational regulation and have additional roles. Our findings indicate that Mos appeared early during animal evolution as an oocyte-expressed kinase and functioned ancestrally in regulating core specializations of female meiosis.

Cnidarian bleaching is a breakdown in the mutualistic symbiosis between host Cnidarians, such as reef building corals, and their unicellular photosynthetic dinoflagellate symbionts. Bleaching is caused by a variety of environmental stressors, most notably elevated temperatures associated with global climate change in conjunction with high solar radiation, and it is a major contributor to coral death and reef degradation. This review examines the underlying cellular events that lead to symbiosis dysfunction and cause bleaching, emphasizing that, to date, we have only some pieces of a complex cellular jigsaw puzzle. Reactive oxygen species (ROS), generated by damage to both photosynthetic and mitochondrial membranes, is shown to play a central role in both injury to the partners and to inter-partner communication of a stress response. Evidence is presented that suggests that bleaching is a host innate immune response to a compromised symbiont, much like innate immune responses in other host-microbe interactions. Finally, the elimination or exit of the symbiont from host tissues is described through a variety of mechanisms including exocytosis, host cell detachment and host cell apoptosis.

Proliferative gill disease (PGD), caused by the myxozoan parasite Henneguya ictaluri, is the most prevalent parasitic infection affecting commercial channel catfish (Ictalurus punctatus) aquaculture. There are currently no effective chemotherapeutic or biological control measures for PGD, which often peaks during the spring and fall when water temperatures are between 16-25 degrees C. The current diagnostic techniques of gross examination of gill clip wet mounts and histopathology are subject to false-negatives during the early stages of infection, and the quantifiable nature of end-point polymerase chain reaction (PCR) is subjective. Consequently, a rapid and more sensitive quantitative real-time PCR assay was developed for the detection of H. ictaluri during the early stages of infection in channel catfish. A 23 base-pair TaqMan probe was designed based on previously published H. ictaluri PCR protocols. The sensitivity of the assay was the equivalent of a single H. ictaluri actinospore, and in a pond challenge study, quantitative real-time PCR proved to be more sensitive than gross examination, microscopic examination of gill clip wet mounts, and histopathologic examination of gill tissue sections. Future applications of this assay will focus on developing methodologies to be used in conjunction with current pond-monitoring protocols to evaluate potential treatments and better manage this significant seasonal disease.

Nitric oxide (NO) is a small molecule with unconventional properties. It is found in organisms throughout the phylogenetic scale, from fungi to mammals, in which it acts as an intercellular messenger of main physiological events, or even as an intracellular messenger in invertebrates. In both vertebrates and invertebrates, NO is involved in many processes, regulated in part by cyclic guanosine monophosphate (cGMP), and reacts with different oxygen molecular species. The presence of NO in the early-diverging metazoan phylum of Cnidaria, of which Hydra represents the first known species having a nervous system, supports a role of this molecule as an ancestral neural messenger with physiological roles that remain to be largely elucidated. Therefore, our novel findings on the presence of NO in Hydra are here integrated in such a comparative frame.

Four new Ceratomyxa species were described from the gallbladder of four Red Sea fishes at Suez and Hurghada, Egypt. These species are Ceratomyxa bassoni sp. n. from Plectorhinchus gaterinus (Forsskal 1775) at Suez and Hurghada, Ceratomyxa entzerothi sp. n. from Valamugil seheli (Forsskal 1775) at Suez and Hurghada, Ceratomyxa swaisi sp. n. from Saurida undosquamis (Richardson 1848) at Suez only and Ceratomyxa hurghadensis sp. n. from Fistularia commersonii Ruppell 1838 at Hurghada only. Their taxonomic affinities to other species are discussed.

Myxobolus dogieli Bykhovskaya-Pavlovskaya & Bykhovski, 1940 is regarded as a site specific myxosporean, infecting the heart of cyprinid fish. During a survey of the myxosporean fauna of Lake Balaton fish, heart myxobolosis was found in the common bream, Abramis brama, with heavy infection of the ventricle and the bulbus arteriosus in some infected bream. Developing and mature plasmodia were mostly in the connective tissue of the subepicardium and subendocardium. Plasmodia developing in the subendocardium protruded into the lumen of the heart, while plasmodia developing in the subepicardium protruded over the epicardium forming large sausage-like outgrowths. Plasmodia with mature spores were found in the summer. The shape and size of the spores corresponded to those of the original description. Phylogenetic analysis based on the 18S rDNA sequence of M. dogieli showed that this species fit well in the genus Myxobolus. As no molecular data are available on spores from the type host, common carp, the species studied by us is temporarily designated as Myxobolus s.l. dogieli.

Little is known about the viability of myxozoan actinospore stages after harvest from laboratory cultures of infected oligochaete worms. The viability and reactivity of actinospores of three myxozoan species was evaluated after short-term storage at 4 degrees C and 12 degrees C. Two methods of determining actinospore viability were compared: differential fluorescent staining and direct microscopic observation of morphological indicators of spore integrity. Spore reactivity was quantified by measuring polar filament discharge rates in a micro-assay with fish mucus substrate and mechanical stimulation by vibration. The age-dependent viability of the three species showed clear differences. Myxobolus cerebralis actinospores had the shortest effective life span whereas Henneguya nuesslini actinospores survived significantly longer. Storage at lower temperatures yielded higher viability in all species. Myxobolus pseudodispar actinospores were significantly robust up to 12 degrees C when assessed by staining, but showed similar viability characteristics as H. nuesslini when analyzed morphologically. Evaluation of spore viability by fluorescent staining correlated with morphological assessment, although fewer viable actinospores were usually detected microscopically. Polar filament discharge activity of morphologically intact actinospores did not significantly decrease until the third day of storage compared to freshly harvested samples. The results indicate that durability and reactivity trends during storage of actinospores differ among myxozoan species.

Henneguya rondoni n. sp. found in the peripheral lateral nerves located below the two lateral lines of the fish Gymnorhamphichthys rondoni (Teleostei, Rhamphichthyidae) from the Amazon river is described using light and electron microscopy. Spherical to ellipsoid cysts measuring up to 110 microm in length contained only immature and mature spores located in close contact with the myelin sheaths of the nervous fibres. Ellipsoidal spores measured 17.7 (16.9-18.1)-microm long, 3.6 (3.0-3.9)-microm wide, and 2.5 (2.2-2.8)-microm (n=25) thick. The spore body measuring 7.0 (6.8-7.3)-microm long was formed by two equal symmetric valves, each with an equal tapering tail 10.7 (10.3-11.0) microm in length. The tails were composed of an internal dense material surrounded by an external homogeneous sheath of hyaline substance. The valves surrounded two equal pyriform polar capsules measuring 2.5 (2.2-2.8)-microm long and 0.85 (0.79-0.88)-microm (n=25) wide and a binucleated sporoplasm cell containing globular sporoplasmosomes 0.38 (0.33-0.42) microm (n=25) in diam. with an internal eccentric dense structure with half-crescent section. Each polar capsule contains an anisofilar polar filament with 6-7 turns obliquely to the long axis. The matrix of the polar capsule was dense and the wall filled with a hyaline substance. The spores differed from those of previously described species. Based on the ultrastructural morphology of the spore and specificity to the host species, we propose a new species name H. rondoni n. sp.

The existence and localisation of carbohydrate terminals in Enteromyxum scophthalmi stages was investigated at light (LM) and transmission electron microscopes (TEM) using lectin histochemistry techniques, with the aim of contributing to elucidate the participation of carbohydrate-lectin interactions in the parasite invasion and relationships with the fish host. The presence of abundant mannose and/or glucose residues was demonstrated by the intense staining by concanavalin A at both LM and TEM. The staining pattern obtained with soybean agglutinin and Bandeiraea simplicifolia I (BSI) indicated the abundance of N-acetyl-galactosamine and D-galactose at a lesser extent. The lectins wheat germ agglutinin, BSI and Ulex europaeus agglutinin produced weaker marks. Most lectins recognised structures present in both pre-sporogonic and sporogonic stages, though the glycosidic pattern and/or staining intensity varied between developmental stages. No staining was obtained with Sambucus nigra agglutinin. The TEM studies demonstrated glucose-mannose, N-acetyl-glucosamine, N-acetyl-galactosamine and alpha-D-galactose as dominant structures at the parasite membrane and host-parasite interface, suggesting a role in host-parasite interactions. All these terminals were also detected in the mitochondria of P cells and were scant in the S cells and nuclei. In turbot intestine, mannose-glucose terminals and N-acetyl-glucosamine were labelled on the epithelial brush border and in the mucous cells and rodlet cells. The relevance of these findings in relation to the host-parasite interaction is discussed.

Nematogenesis, the production of stinging cells (nematocytes) in Cnidaria, can be considered as a model neurogenic process. Most molecular data concern the freshwater polyp Hydra, in which nematocyte production is scattered throughout the body column ectoderm, the mature cells then migrating to the tentacles. We have characterized tentacular nematogenesis in the Clytia hemisphaerica hydromedusa and found it to be confined to the ectoderm of the tentacle bulb, a specialized swelling at the tentacle base. Analysis by a variety of light and electron microscope techniques revealed that while cellular aspects of nematogenesis are similar to Hydra, the spatio-temporal characteristics are markedly more ordered. The tentacle bulb nematogenic ectoderm (TBE) was found to be polarized, with a clear progression of successive nematoblast stages from a proximal zone (comprising a majority of undifferentiated cells) to the distal end where the tentacle starts. Pulse-chase labelling experiments demonstrated a continuous displacement of differentiating nematoblasts towards the tentacle tip, and that nematogenesis proceeds more rapidly in Clytia than in Hydra. Compact expression domains of orthologues of known nematogenesis-associated genes (Piwi, dickkopf-3, minicollagens and NOWA) were correspondingly staggered along the TBE. These distinct characteristics make the Clytia TBE a promising experimental system for understanding the mechanisms regulating nematogenesis.

We have firstly investigated the toxicological activity by hemolytic assay of crude extract obtained by sonication of holotrichous isorhiza isolated nematocysts of the Scyphozoan Pelagia noctiluca, collected in the Strait of Messina. The hemolytic activity was both time- and dose-dependent on fish, rabbit, chicken and human red blood cells. At lowest doses rabbit and chicken erythrocytes were the most sensitive, whereas those of eel were the most resistant to the crude extract. Different storage conditions, such as -20 degrees C, -80 degrees C for up to 6 months and lyophilization, did not affect the stability of crude venom. Moreover, neither treatment at 4 degrees C, 20 degrees C and 37 degrees C for different time periods ranging between 30 min and 24 h, nor harsh thermal treatment at 80 degrees C and 100 degrees C affected the hemolytic power. The crude venom resulted even stable towards proteolysis and alkaline pH values.

The aim of this study has been the biochemical demonstration of the presence of gamma-aminobutyric acid (GABA) in the Mediterranean sea fan Eunicella cavolini by means of high-performance liquid chromatography, and the description of the distribution pattern of GABA and its related molecules, glutamic acid decarboxylase (GAD), vesicular GABA transporter (VGAT) and one of the GABA receptors (GABA(B) R) by immunohistochemical methods. The interrelationships of GABA, GAD and GABA receptor immunoreactivity have been established by using double-immunohistochemical methods and confocal microscopy. The immunodetection of monoclonal and/or polyclonal antibodies has revealed GABA immunoreactivity throughout the polyp tissue, both in neuronal and non-neuronal elements. GAD immunoreactivity has been mostly localized in the neuronal compartment, contacting epithelial and muscular elements. GABA(B) R immunoreactivity appears particularly intense in the nematocytes and in the oocyte envelope; its presence in GAD-immunoreactive neurons in the tentacles suggests an autocrine type of regulation. Western blot analysis has confirmed that a GABA(B) R, with a molecular weight of 142 kDa, similar to that of rat brain, is present in E. cavolini polyp tissue. The identification of the sites of the synthesis, vesicular transport, storage and reception of GABA strongly suggests the presence of an almost complete set of GABA-related molecules for the functioning of the GABAergic system in this simple nervous system. The distribution of these different immunoreactivities has allowed us to hypothesize GABA involvement in nematocyst discharge, in body wall and enteric muscular contraction, in neuronal integration and in male gametocyte differentiation.

Two species of venomous pelagic cnidaria are compared according to their enzymatic, cytotoxic and haemolytic potency. The widely distributed jellyfish Cyanea capillata and Cyanea lamarckii were collected in the North Sea at the coasts of the Orkney Island and the Island of Helgoland. Purified cnidocyst extracts from fishing and mesenteric tentacles were prepared and tested for their bioactivity. The haemolysis induced by toxins of C. capillata was determined with respect to organism size and toxigenic organs. The haemolytic activity of the related species C. lamarckii was documented for the first time. Dose dependent haemolytic activities have been detected by means of protein equivalents at concentrations above 20mug(protein)/mL. Extracts of fishing tentacle cnidocysts showed a less potent haemolytic activity compared to extracts of mesenteric tentacles. In vitro studies with permanent cells of a hepatoma cell line have shown a time and concentration dependent loss of cell vitality up to 90% at 33.3mug(protein)/mL (10mug(protein)/10(5) cells). Supplementing the cell based toxicity tests an enzyme assay was performed to measure a phospholipase A(2) (PLA(2)) activity. A PLA(2)-like activity could be demonstrated in cnidocysts extracts prepared from mesenteric and fishing tentacles of both jellyfish species.

Fossilized embryos from the late Neoproterozoic and earliest Phanerozoic have caused much excitement because they preserve the earliest stages of embryology of animals that represent the initial diversification of metazoans. However, the potential of this material has not been fully realized because of reliance on traditional, non-destructive methods that allow analysis of exposed surfaces only, and destructive methods that preserve only a single two-dimensional view of the interior of the specimen. Here, we have applied synchrotron-radiation X-ray tomographic microscopy (SRXTM), obtaining complete three-dimensional recordings at submicrometre resolution. The embryos are preserved by early diagenetic impregnation and encrustation with calcium phosphate, and differences in X-ray attenuation provide information about the distribution of these two diagenetic phases. Three-dimensional visualization of blastomere arrangement and diagenetic cement in cleavage embryos resolves outstanding questions about their nature, including the identity of the columnar blastomeres. The anterior and posterior anatomy of embryos of the bilaterian worm-like Markuelia confirms its position as a scalidophoran, providing new insights into body-plan assembly among constituent phyla. The structure of the developing germ band in another bilaterian, Pseudooides, indicates a unique mode of germ-band development. SRXTM provides a method of non-invasive analysis that rivals the resolution achieved even by destructive methods, probing the very limits of fossilization and providing insight into embryology during the emergence of metazoan phyla.

This review analyzes activities in the field of marine invertebrate cell culture during the years 1999 to 2004 and compares the outcomes with those of the preceding decade (1988 to 1998). During the last 5 years, 90 reports of primary cell culture studies of marine organisms belonging to only 6 taxa (Porifera, Cnidaria, Crustacea, Mollusca, Echinodermata, and Urochordata) have been published. This figure represents a 2-fold increase in the annual number of publications over the decade 1988 to 1998. Three other trends distinguish the two reviewed periods. First, in recent years studies attempting to improve cell culture methodologies have decreased, while interest in applications of already existing methodologies has increased. This reflects the effects of short-term cultures in attracting new researchers and scientific disciplines to the field. Second, only 17.8% of the recent publications used long-term cultures, compared with 30.0% of the publications in the previous decade. Third, during recent years research in cell cultures has studied fewer model species more extensively (mainly, Botryllus schlosseri, Crassostrea, Mytilus, Penaeus, and Suberites domuncula), signifying a shift from previous investigations that had studied a more diverse range of organisms. From 1988 to 1998 the phylum Mollusca was the most studied taxon (34.4%), but recent years have seen more studies of Porifera and Crustacea (30.0% and 32.2% of publications) than of Mollusca (21.1%). Still, not even a single established cell line from any marine invertebrate has yet been made available. However, the use of new cellular, genomic, and proteomic tools may fundamentally change our strategy for the development of cell cultures from marine invertebrates.

The present review analyses cytomorphological characters of the parasitic cnidarian Polypodium hydriforme, discriminating between those of bilateral (triploblastic) animals, common characters shared with the Myxozoa, and the unique characters of this species. Phylogenetic position of the group of parasitic cnidarians and of the class Polypodiozoa is discussed. A conclusion is made that the cytomorphological characters as well as 18S rDNA analysis of P. hydriforme and Myxozoa justify establishment of a new taxonomic group (a clade) of parasitic cnidarians (Endocnidozoa) uniting Polypodiozoa and Myxozoa (Zrzavý, Hypsa, 2003). The unique characters of P. hydriforme suggest that the phylum Cnidaria is more diverse than commonly supposed, and that P. hydriforme is not an aberrant cnidarian species but a relic organism, which might originally belong to the cnidarian class Polypodiozoa, which underwent reduction in the course of adaptation to parasitism.

Animal eyes have long served as a classical example of independent origin followed by convergence of structures onto a few different solutions. During the past decade this view has been challenged by the discovery of shared developmental regulatory genes. The Pax6 gene in particular is almost universally employed for eye formation in bilaterian animals, despite widely different embryological origins. The resulting controversy on the multiple or single origins of animal eyes has gradually been sharpened by continuing discoveries of further general similarities in the genetic regulatory circuits of eye development. Recent work on gene expression in specified cell types, together with comparative studies of developmental genes in cnidarians, now show some promise to a solution of the controversy.

KPNAYKGKLPIGLWamide, a novel member of the GLWamide peptide family, was isolated from Hydra magnipapillata. The purification was monitored with a bioassay: contraction of the retractor muscle of a sea anemone, Anthopleura fuscoviridis. The new peptide, termed Hym-370, is longer than the other GLWamides previously isolated from H. magnipapillata and another sea anemone, A. elegantissima. The amino acid sequence of Hym-370 is six residues longer at its N-terminal than a putative sequence previously deduced from the cDNA encoding the precursor protein. The new longer isoform, like the shorter GLWamides, evoked concentration-dependent muscle contractions in both H. magnipapillata and A. fuscoviridis. In contrast, Hym-248, one of the shorter GLWamide peptides, specifically induced contraction of the endodermal muscles in H. magnipapillata. This is the first case in which a member of the hydra GLWamide family (Hym-GLWamides) has exhibited an activity not shared by the others. Polyclonal antibodies were raised to the common C-terminal tripeptide GLWamide and were used in immunohistochemistry to localize the GLWamides in the tissue of two species of hydra, H. magnipapillata and H. oligactis, and one species of sea anemone, A. fuscoviridis. In each case, nerve cells were specifically labeled. These results suggest that the GLWamides are ubiquitous among cnidarians and are involved in regulating the excitability of specific muscles.

The paper provides data on the invasion of Acipenser ruthenus (sterlet) in the North Dvina River by Polypodium hydriforme (Cnidaria). Prevalence and intensity of invasion proved to be similar for two fishing sites. Prevalence of invasion exceeded 88%, thus being exceptionally high for P. hydriforme. Intensity of invasion was from 1 to 436 eggs per female. A verage percentage of infected eggs was about 1%.