Development and population growth of Hydra viridissima Pallas, 1766 (Cnidaria, Hydrozoa) in the laboratory

Hydras, the most representative freshwater Cnidaria, are of common occurrence in bodies of water in every continent except Antarctica. This study was planned with the aim of maintaining a population of Hydra viridissima in laboratory culture to enable the determination of the individual and population growth-rates of this species, as well as its population doubling time and generation time, with a view to employing these common animals as test-organisms in ecotoxicological assays. The organisms were maintained in reconstituted water at 20 +/- 2 degrees C, illuminated at 800 lux with a photoperiod of 12 hours light: 12 hours dark, and were fed on neonates of the cladoceran Ceriodaphnia silvestrii (3 or 4 neonates per hydra, 3 times a week). The individual growth-rate (k) of the species was 0.43, the maximum length of the column 2.53 mm and the generation time 6.6 +/- 1.5 days on average. The hydra population showed an intrinsic growth-rate (r) of 0.0468, according to the fitted curve, and a doubling time of 14.8 +/- 2.63 days. Hydra viridissima is easy to grow in the laboratory and performs well in the conditions used in this study. It is thus a promising candidate test-organism for ecotoxicological studies.     

Morphology and morphodynamics of the stenotele nematocyst of Hydra attenuata Pall. (Hydrozoa,Cnidaria)

This light- and electron-microscopic study has investigated the structure, the morphodynamics of discharge, and the impact of the stenotele cyst of Hydra attenuata (Hydrozoa, Cnidaria) on the prey's integument. The triggered capsule, which is ejected from the cell, discharges its tubular content (shaft, stylets and tubule) by a process of evagination. In doing so the three joined stylets punch a hole into the cuticle of the prey through which the long evaginating tubule penetrates into the interior of the target. The behaviour of the tubule is described in detail and the functional significances of the various parts of the capsule are discussed.     

Coordinated modulation of cellular signaling through ligand-gated ion channels in Hydra vulgaris (Cnidaria, Hydrozoa)

Cnidarians lack well developed organs, but they have evolved the molecular and cellular components needed to assemble a nervous system. The apparent 'simplicity' of the cnidarian nervous net does not occur at the cellular level, but rather in the organisation of conducting systems. Cnidarian neurons are in fact electrically excitable, show the typical extended morphology and are connected by chemical synapses or gap junctions. They have been regarded as peptidergic, given the wealth of neuropeptides generally distributed along neurites and in cell bodies, supporting the hypothesis of a modulatory role in neurotransmission. However, the presence of clear-cored, as well as dense-cored synaptic vesicles in cnidarian neurons suggests both fast and slow synaptic transmission mechanisms. In fact, biochemical and functional evidence indicates that classical neurotransmitters and their metabolic partners are present in cnidarian tissues, where they are involved in coordinating motility and behavior. We have identified and characterized in Hydra tissues receptors to the inhibitory and excitatory amino acid neurotransmitters, GABA, glycine and NMDA, that are similar to mammalian ionotropic receptors in terms of their biochemical and pharmacological properties. These receptors appear to regulate pacemaker activities and their physiological correlates; in the live animal, they also affect feeding behavior, namely the duration and termination of the response elicited by reduced glutathione, with opposite actions of GABA and glycine or NMDA, respectively. These results suggest that modulation of cellular signaling through ligand-gated-ion channels is an ancient characteristic in the animal kingdom, and that the pharmacological properties of these receptors have been highly conserved during evolution.     

Poly(gamma-glutamic acid)s are the major constituents of nematocysts in Hydra (Hydrozoa, Cnidaria).

The 2.80 +/- 0.20 mumol of anions found/mg of isolated and purified dry nematocysts (capsular secretory products of stinging cells) from Hydra make up the majority of the soluble capsular content. They are, in cooperation with corresponding cations, responsible for the generation and regulation of an internal osmotic pressure that amounts up to 150 bar (Weber, J. (1989) Eur. J. Biochem. 184, 465-476). The anions are organized as linear homopolymers of L-glutamic acids which are linked by gamma-carboxyl-alpha-amino amide bonds; the degree of polymerization is heterogeneous and dependent on the particular type of nematocyst. In situ the intracapsular glutamic acid monomer concentration is as high as 2 M. This is the first time that poly(gamma-glutamic acid)s, which are known to occur in some selected bacteria, are reported for eucaryotes. It is suggested that they may also be present as predominant components in nematocysts of other cnidarian species and thus might represent a class of compounds which is characteristic for a whole phylum of the animal kingdom.     

Different roles of GABA and glycine in the modulation of chemosensory responses in Hydra vulgaris (Cnidaria,Hydrozoa)

Phylogenetic studies suggest that GABA and glycine receptors derive, as a result of divergent evolution, from a common ancestral protoreceptor originated in a unicellular organism. This raises the possibility that members of the ligand-gated ion channels (LGIC) superfamily might be widely present in living organisms including bacteria and primitive invertebrates. High-affinity GABA receptors occur in the tissues of Hydra vulgaris whose pharmacological characteristics compare with those of mammalian ionotropic GABA receptors. Behavioural studies have shown that activation of these GABA _A -like receptors by their allosteric modulators increases the duration of response to reduced glutathione (GSH). Recently, strychnine-sensitive glycine receptors have been shown to occur in Hydra tissues. Activation of these glyR also results in increased duration of the response to GSH. In order to investigate the contribution of endogenous transmitters to the modulation of the feeding response, we studied the effects of exposing the polyps to brief depolarizing pulses prior to the GSH test. A severe inhibition of the response was observed following exposure to KCl or veratridine. Administration of GABA or muscimol counteracted the effects of the pulses in a dose-dependent manner. The effects of GABA or muscimol were suppressed by the GABA _A -specific antagonist gabazine both in pulse-untreated and treated polyps. By contrast, glycine and its agonist taurine were not able to restore the physiological duration of response in pulse-treated Hydra, while another glyR agonist, β-alanine, partially reduced the pulse-induced inhibition. We conclude that GABA appears to be the major inhibitory transmitter responsible for the regulation of the feeding response. Molecular studies aimed at identifying GABA receptor subunits are in progress.     

First record of sympagic hydroids (Hydrozoa,Cnidaria) in Arctic coastal fast ice

We report on the first record of interstitial cnidarians in sea ice. Ice core samples were collected during eight field periods between February 2003 and June 2006 in the coastal fast ice off Barrow, Alaska (71°N, 156°W) at four locations. A total of 194 solitary, small (0.2–1.1 mm) elongated specimens of a previously unknown interstitial hydroid taxon were found. By cnidome composition and the occurrence of a highly retractable pedal disc formed by epidermal tissue only, the specimens are tentatively assigned to representatives of the family Protohydridae, subclass Anthomedusae. The hydroids were found almost exclusively in the bottom 10 cm-layer (at the ice–water interface) of 118 ice cores, with abundances ranging from 0 to 27 individuals per core section (0–4,244 ind m⁻²) and a grand mean of 269 ind m⁻² in bottom 10 cm-layer sections. Abundances were lower in December and late May than in months in between with considerable site variability. A factor analysis using 12 variables showed that hydroid abundance correlated highest with abundances of copepod nauplii and polychaete juveniles suggesting a trophic relationship.     

Cholinergic mechanism in Liriope tetraphylla (Cnidaria, Hydrozoa)

Crude whole body homogenates of Liriope tetraphylla exhibit a cholinesterase particularly active on acetylthiocholine but not on butyrylthiocholine. The acetylthiocholine hydrolysis is completely blocked by neostigmine. The Michaelis-Menten constant for acetylthiocholine is 0.14 mM. The pharmacological analysis of the responses to the choline esters nicotine and atropine suggests the involvement in Liriope tetraphylla of a cholinergic mechanism in the pointing reflex. Butyrylcholine, nicotine and atropine (but not muscarinic agonists) caused the contraction of the subumbrellar radial muscles. The effects of atropine were dose-dependent and were depressed in competition with muscarinic agonists. MgCl2 interfered with the action of atropine. The results were explained by suggesting the existence, at least at the neuromuscular junction, of excitatory (nicotinic) and inhibitory (muscarinic) pre-synaptic receptors modulating the release of the (unknown) transmitter acting post-synaptically.     

Character evolution in Hydrozoa (phylum Cnidaria)

The diversity of hydrozoan life cycles, as manifested in the wide range of polyp, colony, and medusa morphologies, has been appreciated for centuries. Unraveling the complex history of characters involved in this diversity is critical for understanding the processes driving hydrozoan evolution. In this study, we use a phylogenetic approach to investigate the evolution of morphological characters in Hydrozoa. A molecular phylogeny is reconstructed using ribosomal DNA sequence data. Several characters involving polyp, colony, and medusa morphology are coded in the terminal taxa. These characters are mapped onto the phylogeny and then the ancestral character states are reconstructed. This study confirms the complex evolutionary history of hydrozoan morphological characters. Many of the characters involving polyp, colony, and medusa morphology appear as synapomorphies for major hydrozoan clades, yet homoplasy is commonplace.     

Vibrio harveyi Associated with Aglaophenia octodonta (Hydrozoa, Cnidaria)

A previously unknown association between a luminous bacterium, Vibrio harveyi, and a benthic hydrozoan, Aglaophenia octodonta, is described. Aglaophenia hydrocladia showed a clear fluorescence in the folds along the hydrocaulus and at the base of the hydrotheca, suggesting the presence of luminous bacteria. This hypothesis was confirmed by isolation of luminous bacteria from Aglaophenia homogenates. Phenotypic characterization of bacterial isolates was performed by several morphological, biochemical, and cultural tests, completed with 16S rDNA sequence analysis. All the isolates were referred to a single species: V. harveyi. The association between V. harveyi and A. octodonta has epidemiological as well as ecological significance. Therefore, A. octodonta may function as habitat “islands” providing a unique set of environmental conditions for luminous bacteria colonization, quite different from those already recorded from the plankton for other Vibrio species.     

Species in the genus Turritopsis (Cnidaria, Hydrozoa): a molecular evaluation

Mitochondrial ribosomal gene sequences were used to investigate the status of several populations of hydromedusae belonging to the genus Turritopsis (family Oceaniidae). Several nominal species have been described for this genus, but most of them had been synonymized and attributed to one cosmopolitan species, Turritopsis nutricula . A recent revision based on morphological and reproductive characters, however, has shown that many different populations can be distinguished and that several of the nominal Turritopsis species are likely valid biological species. Our investigation using molecular sequence data of 16S mitochondrial gene confirms these results. The Mediterranean Turritopsis must be attributed to Turritopsis dohrnii and the Turritopsis of New Zealand must be referred to Turritopsis rubra . The situation of the Japanese Turritopsis is more complex, though all sampled populations are clearly distinct from T. nutricula , a species likely confined to the Western Atlantic. The Japanese Turritopsis fall into three widely separated lineages. One of them, corresponding likely to Turritopsis pacifica , is closely related to T. rubra . A second clade, which potentially represents an as yet undescribed species, produces smaller medusae than T. pacifica and is morphologically distinguishable from it. Finally, a third group was distinguished by a single haplotype sequence that is identical with a Mediterranean sample of T. dohrnii . It is postulated that the last group of Japanese Turritopsis is likely a recent introduction, most probably by human activity. A survey of all known and potentially valid Turritopsis species is given in table format to facilitate identifications and future revisory work.     

Prospective fate of early blastomeres in Hydractinia echinata (Cnidaria,Hydrozoa)

Summary Blastomeres of two-cell, four-cell, and eight-cell embryos of Hydractinia echinata were injected with horseradish-peroxidase (HRP) or fluorescein isothiocyanate (FITC)-dextran. The fate of the descendants of the injected blastomeres was followed until the planula larva had developed. The results obtained after HRP or FITC-dextran injection were essentially the same. Blastomeres are equivalent up to the four-cell stage, i.e. half-blastomeres produce half of the ectoderm of the planula larva and quarter-blastomeres give rise to one quarter of the larval ectoderm. During normal embryogenesis, the larval anterior-posterior axis corresponds to the animal-vegetal axis of the zygote. Thus, the labelled areas of larvae consisting of the progeny of injected half or quarter blastomeres normally stretch along the larval anterior-posterior axis. Normally, material giving rise to anterior or posterior larval parts, respectively, is separated at the third cleavage. Irrespective of the type of experiment, the progeny of injected blastomeres always contributed to endoderm formation, i.e. in larvae resulting from injected embryos the endoderm was more or less uniformly labelled. Application of vital stains locally to the exterior of zygotes and following these markers through first and second cleavage, produced evidence that in the vast majority of cases, the second cleavage is meridional.Offprint requests to: A. Schlawny     

Glycosaminoglycans in Hydra magnipapillata (Hydrozoa, Cnidaria): demonstration of chondroitin in the developing nematocyst, the sting organelle, and structural characterization of glycosaminoglycans

The hydrozoan is the simplest organism whose movements are governed by the neuromuscular system, and its de novo morphogenesis can be easily induced by the removal of body parts. These features make the hydrozoan an excellent model for studying the regeneration of tissues in vivo, especially in the nervous system. Although glycosaminoglycans (GAGs) and proteoglycans (PGs) have been implicated in the signaling functions of various growth factors and play critical roles in the development of the central nervous system, the isolation and characterization of GAGs from hydrozoans have never been reported. Here, we characterized GAGs of Hydra magnipapillata. Immunostaining using anti-GAG antibodies showed chondroitin or chondroitin sulfate (CS) in the developing nematocyst, which is a sting organelle specific to cnidarians. The CS-PGs might furnish an environment for assembling nematocyst components, and might themselves be components of nematocysts. Therefore, GAGs were isolated from Hydra and their structural features were examined. A considerable amount of CS, three orders of magnitude less heparan sulfate (HS), but no hyaluronan were found, as in Caenorhabditis elegans. Analysis of the disaccharide composition of HS revealed glucosamine 2-N-sulfation, glucosamine 6-O-sulfation, and uronate 2-O-sulfation. CS contains not only nonsulfated and 4-O-sulfated N-acetylgalactosamine (GalNAc) but also 6-O-sulfated GalNAc. The average molecular size of CS and HS was 110 and 10 kDa, respectively. It has also been established here that CS chains are synthesized on the core protein through the ubiquitous linkage region tetrasaccharide, suggesting that indispensable functions of the linkage region in the synthesis of GAGs have been conserved during evolution.     

Benthic hydroids (Cnidaria,Hydrozoa) from the Balleny Islands (Antarctica)

Twenty-two species of benthic hydroids, belonging to ten families and 14 genera, were found in a hydroid collection obtained in the Balleny Islands during the BioRoss expedition with the NIWA research vessel Tangaroa in 2004. Twenty of those species constitute new records for the Balleny Islands, raising the total number of known species in the area to 25. Most are members of the subclass Leptothecata, although the subclass Anthoathecata is also relatively well represented. Kirchenpaueriidae and Sertulariidae constitute families with the greatest numbers of species in the collection, with five species (20%) each. Oswaldella with five species (20%) and Staurotheca with four (16%), were the most diverse genera. Twelve species (63%) are endemic to Antarctic waters, most of them with a circum-Antarctic distribution, and 17 (89%) are restricted to Antarctic or Antarctic/sub-Antarctic waters. Although the Balleny Islands hydroid fauna seems to be a typical Antarctic assemblage, it has some striking peculiarities, namely the absence or low representation of some typical and widespread Antarctic genera (Antarctoscyphus and Schizotricha/Symplectoscyphus, respectively).     

Benthic hydroids (Cnidaria, Hydrozoa) from off Bouvet Island (Antarctic Ocean)

Twenty-seven species of benthic hydroids have been found in a small collection from off Bouvet Island (Antarctic Ocean). The material was obtained during the XXI/2 German Antarctic expedition with R.V. Polarstern 2003–2004. Of the 27 species collected, 21 were unknown from those waters. Consequently, the number of known species of benthic hydroids from the area has risen to 32, representing a high diversity location for this group in the Southern Ocean considering the sampling effort made. All species, including those previously known from Bouvet waters, are considered with respect to geographical distribution.The present work is an addition to the special issue “Macrobenthic studies at Bouvet island”, presenting work carried out from on board RV Polarstern in 2003/2004 and published in Polar Biology 29 (2006)     

Structure and function of the locomotory system ofPolyorchis montereyensis (Cnidaria,Hydrozoa)

Summary 1. The structure and function of the locomotory system of the anthomedusanPolyorchis montereyensisSkogsberg were studied in detail. Anatomical investigations were carried out primarily on fresh or formalin fixed specimens; histology was done on specimens fixed in Bouin's fluid. Functional analyses were based largely on photography and cinematography.2. Swimming inP. montereyensis involves the alternating antagonistic action of the subumbrellar swimming muscles and the elastic mesoglea.3. The swimming muscle consists of striated contractile elements arranged circularly in four discontinuous subumbrellar sheets and a sheet on the subumbrellar side of the velum. There is also a sheet of radially arranged fibers on the exumbrellar side of the velum. The four subumbrellar sheets are anchored to the bell along the perand interradii.4. The mesogleal skeleton consists of five components: (a) the matrix of the bell mesoglea, (b) optically visible fibers that traverse the bell from gastrodermal lamella to exumbrella, (c) the basement membrane or supporting lamella, (d) a system of joints, and (e) the velar mesoglea.5. The morphology, orientation, and distribution of the mesogleal fibers suggest that their major role is maintaining the radial integrity of the bell during deformation. The amount of stretch in a region of the bell wall during contraction is inversely proportional to the number of fibers per unit area there. In regions of the bell which are not deformed during contraction fibers are sparse or absent.6. Mesogleal volume remains constant during swimming. Locally the mesoglea is subjected to forces of stretch and compression, but the critical element in narrowing the bell involves bending or folding the mesoglea around a series of structural joints. The fulcrum of these joints is anchored to the exumbrella by concentrations of mesogleal fibers. The joints consist of eight adradial regions of highly deformable mesoglea lacking visible fibers. The regions are triangular in cross section and are separated from the remainder of the mesoglea (98–99 % of the total) by the gastrodermal lamella. A circular apical joint is also present.7. Sequential changes in shape and position of the bell relative to a fixed grid during contraction and recovery were measured in order to determine such parameters of swimming as rate of contraction, rate of expulsion of water, change in bell velocity during contraction and recovery, momentum, etc.8. The function of the velum was determined by cinematographic analysis of swimming animals both before and after removal of the velum. In normal swimming the velum serves mainly to constrict the aperture of the bell, thus increasing the velocity of expelled water, and hence increasing the force driving the medusa foward. Medusae swam with a greatly decreased velocity after velum removal.9. Turning is accomplished primarily by asymmetrical contraction of the exumbrellar velar radial muscles, whereby the velar aperture is displaced to one side, water is expelled obliquely, and the bell turns toward that same side. The ability to turn was lost after velum removal.10. Studies of the relationship between individual size and the various parameters of swimming inP. montereyensis show that: (a) the duration of the contraction phase of the swimming beat is roughly proportional to the square root of the subumbrellar circumference (or bell height); (b) smaller individuals swim faster relative to their bell height than do larger ones; (c) the velum is relatively better developed in small animals and plays a proportionately more important role during swimming.

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Struktur und Funktion des lokomotorischen Systems vonPolyorchis montereyensis (Cnidaria, Hydrozoa)

Kurzfassung Mit Hilfe histologischer und kinematographischer Methoden wurden Bau und Funktion des lokomotorischen Systems der AnthomedusePolyorchis montereyensisSkogsberg analysiert. Die Schwimmbewegungen resultieren aus der antagonistischen Wirkung der Muskulatur der Subumbrella und der elastischen Mesogloea. Struktur, Anordnung, Verteilung und Verankerung der Muskelzellen werden beschrieben. Die Körperschicht der Mesogloea besteht aus 5 Komponenten mit Skelettfunktion: der Matrix der Schirmmesogloea, einem System von Muskelfasern, der Stützlamelle, acht adradialen Verbindungssträngen von stark deformierbarer, fibrillenloser Mesogloea und der Mesogloea des Velums. Die Fribrillen und Verbindungsstränge der Mesogloea bewirken, daß die Schirmglocke während der Kontraktion deformiert werden kann. Die Funktion des Velums sowie die kontinuierlichen Veränderungen der Glocke in bezug auf Form und Lage während des Schwimmvorgangs, insbesondere die Geschwindigkeit der Kontraktion, des Wasserausstoßes und der Fortbewegung bei Individuen verschiedener Größe, wurden eingehend untersucht. Kleinere Medusen schwimmen relativ schneller als größere, was hauptsächlich auf stärkere Kontraktionen des Velums zurückzuführen ist.