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Molecular evolution of the Ca(2+)-binding photoproteins of the Hydrozoa.

Alignment of the primary structures of the hydrozoan photoproteins, aequorin, mitrocomin, clytin and obelin showed very strong amino acid sequence identities. The Ca(2+)-binding sites of the proteins were found to be highly conserved. The Ca(2+)-binding sites were also homologous to the Ca(2+)-binding sites of other Ca(2+)-binding proteins. However, aequorin, mitrocomin, clytin and obelin differed from other Ca(2+)-binding proteins in that they contained a relatively large number of cysteine, tryptophan, histidine, proline and tyrosine residues, suggesting that these residues may have evolved as part of the light-emitting mechanism. Construction of a phylogenetic tree showed that aequorin, mitrocomin, clytin and obelin form a closely related group of proteins.

Aequorin↗

Apoptosis--a death-inducing mechanism tightly linked with morphogenesis in Hydractina echinata (Cnidaria, Hydrozoa).

Programmed cell death is not only known as a mechanism mediating tissue destruction, but also as an organismic tool for body shaping and regulation of morphological events during development. Here we report the tight and vital link of the most prominent form of programmed cell death, apoptosis, to one of the oldest, most basic, and most radical developmental processes, the metamorphosis of the marine hydrozoon Hydractinia echinata. Apoptosis, represented by DNA fragmentation, appears very early during metamorphosis, approximately 20 minutes post induction. It is then executed in a very distinct spatial and temporal pattern, including the removal or phagocytosis of a large number of larval cells prior to the appearance of stolons and tentacles. Our data indicate a developmental program striving to reduce all body parts that are no longer necessary, before reaching a distinct turning point, when the development of adult features is initiated. During these events, morphogenesis of basal and apical structures correlates with recycling of that particular larval region, indicated by the presence of apoptosis. Based on these data, the necessity of apoptosis for normal development of adult patterns is inferred and a fundamental association of apoptosis with developmental processes can be stated.

Animals↗

Gap and septate junctions in the excitable endoderm of Polyorchis penicillatus (Hydrozoa, Anthomedusae).

The morphological basis of impulse conduction in a jellyfish epithelium was investigated. Lanthanum impregnation of endodermal canal and endodermal lamella verified the existence of true gap junctions in Polyorchis. In both transverse and en face sections of gap junctions, electron-lucent globules, with a width of 7--8 nm and a spacing of about 11 nm, are evident. Gap-junctions are concentrated at the peripheral canal margin and septate junctions are localized around the canal lumen. Epithelial cells of the endodermal canals are capable of conducting a non-decrementing action potential. It is suggested that endodermal spike propagation, which can mediate 'crumpling' behaviour, is dependent upon low-resistence ionic pathways provided by gap-junctions and upon sealing of the intercellular space from saline extracellular fluids by septate junctions.

Action Potentials↗

Calcification in the planula and polyp of the hydroid Hydractinia symbiolongicarpus (Cnidaria, Hydrozoa).

This study examines calcification in planulae and polyps of the hydroid Hydractinia symbiolongicarpus. We observed that established colonies produce a crystalline mat on their substratum and that crystals visible by polarized light microscopy occur in the vacuoles of the gastrodermal cells of both polyps and planulae. The crystalline mat was found by infrared spectroscopy to contain calcium carbonate in the form of aragonite. The composition of the vacuolar crystals and the cellular mechanisms for manufacturing them were explored by alteration of calcium levels in the environment and by the use of pharmacological agents (acetazolamide, caffeine, DIDS, diltiazem, nifedipine, procaine, Ruthenium Red, ryanodine and verapamil) that affect cellular uptake and transport of calcium and bicarbonate. The results indicated that the crystals in the vacuoles contained calcium carbonate. The gastrodermal cells are hypothesized to serve as a physiological sink for excess calcium that enters the organism during motility, secretion and metamorphosis of the planula, and to create a crystalline substratum for the colony of polyps.

Animals↗

Shaping of colony elements in Laomedea flexuosa Hinks (Hydrozoa, Thecaphora) includes a temporal and spatial control of skeleton hardening.

The colonies of thecate hydroids are covered with a chitinous tubelike outer skeleton, the perisarc. The perisarc shows a species-specific pattern of annuli, curvatures, and smooth parts. This pattern is exclusively formed at the growing tips at which the soft perisarc material is expelled by the underlying epithelium. Just behind the apex of the tip, this material hardens. We treated growing cultures of Laomedea flexuosa with substances we suspected would interfere with the hardening of the perisarc (L-cysteine, phenylthiourea) and those we expected would stimulate it (dopamine, N-acetyldopamine). We found that the former caused a widening of and the latter a reduction in the diameter of the perisarc tube. At the same time, the length of the structure elements changed so that the volume remained almost constant. We propose that normal development involves a spatial and temporal regulation of the hardening process. When the hardening occurs close to the apex, the diameter of the tube decreases. When it takes place farther from the apex, the innate tendency of the tip tissue to expand causes a widening of the skeleton tube. An oscillation of the position at which hardening takes place causes the formation of annuli.

Animals↗

Variations of ATP content in V79 cells treated with crude toxins of--Aequorea aequorea (Cnidaria: Hydrozoa) and Rhizostoma pulmo (Cnidaria: Scyphozoa). A preliminary study.

The toxicity of Cnidaria exerts a noticeable influence on some human activities, such as fishery and bathing, and on public health. As toxins of Mediterranean Cnidaria are located in nematocysts and in tissues, in this study the influence of crude toxins (nematocyst and surrounding tissue venom) extracted from the jellyfish Aequorea aequorea and Rhizostoma pulmo on ATP content of cultured V79 cells was assessed. Using the crude toxin of A. aequorea an increase of ATP levels in treated cells was noted; highest values (41.2 10(-7) mM/ml after 180 min treatment) were reached using the highest dose. Otherwise, a generalized decrease of ATP levels was observed treating cells with crude toxin of R. pulmo; recorded values showed the complete depletion of cell ATP at 115 min treatment with the highest dose. A statistical significance was recorded between treatment times and between doses using crude toxin of R. pulmo, and only between treatment times for A. aequorea.

Adenosine Triphosphate↗

Autoaggressive, multi-headed and other mutant phenotypes in Hydractinia echinata (Cnidaria: Hydrozoa).

In an inbreeding program conducted with the colonial hydroid Hydractinia echinata, each F1 mating produced up to 50% F2 offspring displaying an aberrant, clone-constant phenotype, hence referred to as mutant strain. In autoaggressive strains, in one or several areas of the colony autoreactive stolons direct their aggressive devices (stolon tips filled with cytotoxic stinging cells), normally used to kill allogeneic competitors for living space, towards neighboring stolons or polyps (hydranths) of their own colony. In these areas tumor-like masses of self-aggressive stolons were formed, in severe cases causing the death of the colony. Based on previous genetic studies, the interpretation proposed here attributes autoaggressive behavior to a mosaic-type alternative expression of arl (allorecognition) alleles in heterozygous individuals. Developmental mutant strains termed He-mh form supernumerary heads during regeneration and normal development as well. Common to all He-mh phenotypes isthe production of additional headsalong the bodycolumn of fully-grown polyps. The heads give rise to complete hydranths connected by a tube that derives from the gastric region of the original polyp and eventually transforms into a stolon. In bastol strains, polyps convert the basal region of their body column into a periderm-covered stolon from which the residual apical hydranth detaches. Colonies expressing both the He-mh and the bastol (bst) phenotype frequently lose detaching multi-headed hydranths and the colony disintegrates. The large number of mutant F2 offspring reveals high genetic variability in Hydractinia.

Alleles↗

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.

Amino Acids↗

Control of formation of the two types of polyps in Thecocodium quadratum (Hydrozoa, Cnidaria).

Thecocodium quadratum (Werner, Jber. Biol. Anst. Helgoland, 1965) is a colonial hydroid which produces 2 different types of polyps: gastrogonozooids and dactylozooids. The mouthless dactylozooids bear tentacles and catch the prey, which is then taken over and swallowed by the gastrogonozooids which have no tentacles. It is obvious that for a colony to survive both polyps must exist simultaneously arranged in a certain spatial pattern. Our experiments indicate that the formation of polyps in a growing culture is governed by at least 3 principles: (1) short range inhibition between polyps irrespective of their differentiation; (2) long range specific inhibition between gastrogonozooids; and (3) long range supporting influence (lateral help, Meinhardt, H., Models of Biological Pattern Formation, 1982) between gastrogonozooids and dactylozooids.

Animals↗

Isolation of Hox genes from the scyphozoan Cassiopeia xamachana: implications for the early evolution of Hox genes.

The isolation of Hox genes from two cnidarian groups, the Hydrozoa and Anthozoa, has sparked hypotheses on the early evolution of Hox genes and a conserved role for these genes for defining a main body axis in all metazoan animals. We have isolated the first five Hox genes, Scox-1 to Scox-5, from the third cnidarian class, the Scyphozoa. For all but one gene, we report full-length homeobox plus flanking sequences. Four of the five genes show close relationship to previously reported Cnox-1 genes from Hydrozoa and Anthozoa. One gene, Scox-2, is an unambiguous homologue of Cnox-2 genes known from Hydrozoa, Anthozoa, and also Placozoa. Based on sequence similarity and phylogenetic analyses of the homeobox and homeodomain sequences of known Hox genes from cnidarians, we suggest the presence of at least five distinct Hox gene families in this phylum, and conclude that the last common ancestor of the Recent cnidarian classes likely possessed a set of Hox genes representing three different families, the Cnox-1, Cnox-2, and Cnox-5 families. The data presented are consistent with the idea that multiple duplication events of genes have occurred within one family at the expense of conservation of the original set of genes, which represent the three ancestral Hox gene families.

Amino Acid Sequence↗

Morphological and ultrastructural analysis of Turritopsis nutricula during life cycle reversal.

The hydrozoa life cycle is characterized, in normal conditions, by the alternation of a post-larval benthic polyp and an adult pelagic medusa; however, some species of Hydrozoa react to environmental stress by reverting their life cycle: i.e. an adult medusa goes back to the juvenile stage of polyp. This very uncommon life cycle could be considered as some sort of inverted metamorphosis. A morphological study of different stages during the reverted life cycle of Turritopsis nutricula led to the characterization of four different stages: healthy medusa, unhealthy medusa, four-leaf clover and cyst. The ultrastructural study of the cellular modifications (during the life cycle reversion of T. nutricula) showed the presence of both degenerative and apoptotic processes. Degeneration was prevalent during the unhealthy medusa and four-leaf clover stages, while the apoptotic rate was higher during the healthy medusa and cyst stages. The significant presence of degenerative and apoptotic processes could be related to the occurrence of a sort of metamorphosis when an adult medusa transforms itself into a polyp.

Animals↗

Medusozoan phylogeny and character evolution clarified by new large and small subunit rDNA data and an assessment of the utility of phylogenetic mixture models.

A newly compiled data set of nearly complete sequences of the large subunit of the nuclear ribosome (LSU or 28S) sampled from 31 diverse medusozoans greatly clarifies the phylogenetic history of Cnidaria. These data have substantial power to discern among many of the competing hypotheses of relationship derived from prior work. Moreover, LSU data provide strong support at key nodes that were equivocal based on other molecular markers. Combining LSU sequences with those of the small subunit of the nuclear ribosome (SSU or 18S), we present a detailed working hypothesis of medusozoan relationships and discuss character evolution within this diverse clade. Stauromedusae, comprising the benthic, so-called stalked jellyfish, appears to be the sister group of all other medusozoans, implying that the free-swimming medusa stage, the motor nerve net, and statocysts of ecto-endodermal origin are features derived within Medusozoa. Cubozoans, which have had uncertain phylogenetic affinities since the elucidation of their life cycles, form a clade-named Acraspeda-with the scyphozoan groups Coronatae, Rhizostomeae, and Semaeostomeae. The polyps of both cubozoans and hydrozoans appear to be secondarily simplified. Hydrozoa is comprised by two well-supported clades, Trachylina and Hydroidolina. The position of Limnomedusae within Trachylina indicates that the ancestral hydrozoan had a biphasic life cycle and that the medusa was formed via an entocodon. Recently hypothesized homologies between the entocodon and bilaterian mesoderm are therefore suspect. Laingiomedusae, which has often been viewed as a close ally of the trachyline group Narcomedusae, is instead shown to be unambiguously a member of Hydroidolina. The important model organisms of the Hydra species complex are part of a clade, Aplanulata, with other hydrozoans possessing direct development not involving a ciliated planula stage. Finally, applying phylogenetic mixture models to our data proved to be of little additional value over a more traditional phylogenetic approach involving explicit hypothesis testing and bootstrap analyses under multiple optimality criteria. [18S; 28S; Cubozoa; Hydrozoa; medusa; molecular systematics; polyp; Scyphozoa; Staurozoa.].

Animals↗

Isolation and characterization of a mini-collagen gene encoding a nematocyst capsule protein from a reef-building coral, Acropora donei.

Genomic and cDNA clones of a mcol gene encoding mini-collagen (MCOL), a nematocyst capsule protein, have been isolated from a reef-building coral, Acropora donei (Anthozoa). The gene and its flanking regions, comprising 5382 bp and covering three exons and two introns, were sequenced. Exons 2 and 3 together have an open reading frame which can encode a MCOL of 176 amino acids (aa). The coral MCOL has all the characteristic regions present in the four hydra MCOL specified by the four mcol cDNA clones previously isolated from Hydra magnipapillata (Hydrozoa) by Kurz et al. [J. Cell Biol. 115 (1991) 1159-1169], including a central Gly-Xaa-Yaa region and flanking Pro-rich and Cys-repeat regions. This observation suggests that a mcol family is highly conserved in Anthozoa and Hydrozoa, and also that the characteristic regions present in MCOL are essential for the structure and function of these peptides.

Amino Acid Sequence↗

The Sine oculis/Six class family of homeobox genes in jellyfish with and without eyes: development and eye regeneration.

The development of visual organs is regulated in Bilateria by a network of genes where members of the Six and Pax gene families play a central role. To investigate the molecular aspects of eye evolution, we analyzed the structure and expression patterns of cognate members of the Six family genes in jellyfish (Cnidaria, Hydrozoa), representatives of a basal, non-bilaterian phylum where complex lens eyes with spherical lens, an epidermal cornea, and a retina appear for the first time in evolution. In the jellyfish Cladonema radiatum, a species with well-developed lens eyes in the tentacle bulbs, Six1/2-Cr and Six3/6-Cr, are expressed in the eye cup. Six4/5-Cr is mainly expressed in the manubrium, the feeding, and sex organ. All three Six genes are expressed in different subsets of epidermal nerve cells, possibly of the RFamide type which are part of a net connecting the different eyes with each other and the effector organs. Furthermore, expression is found in other tissues, notably in the striated muscle. During eye regeneration, expression of Six1/2-Cr and Six3/6-Cr is upregulated, but not of Six4/5-Cr. In Podocoryne carnea, a jellyfish without eyes, Six1/2-Pc and Six3/6-Pc are also expressed in the tentacle bulbs, Six1/2-Pc additionally in the manubrium and striated muscle, and Six3/6-Pc in the mechanosensory nematocytes of the tentacle. The conserved gene structure and expression patterns of all Cladonema Six genes suggest broad conservation of upstream regulatory mechanisms in eye development.

Animals↗