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PubMed1989

Nucleotide sequence of an actin-encoding gene from Hydra attenuata: structural characteristics and evolutionary implications.

We have determined the complete nucleotide sequence of an actin-encoding gene from Hydra attenuata as well as partial sequences of cDNA clones from two additional actin-encoding genes. The gene from the genomic clone contains a single intron, and has promoter and polyadenylation signals similar to those found in other species. The hydra genome has a very A + T-rich base composition (71%). This is reflected in the codon usage of the actin-encoding genes, which is strongly biased towards codons having A or T in the third position. The hydra actin-encoding gene family consists of three or more transcribed genes, two of which are very closely related to each other and probably arose by a recent gene duplication. Hydra actin, like other invertebrate actins, is more similar to the non-muscle isotypes of vertebrates than to the vertebrate muscle actins. Hydra actin is more similar to animal actins than to those of plants or fungi, which is consistent with the view that all metazoans arose from a single protist ancestor.

Actins↗
PubMed1994

The role of the cAMP pathway in mediating the effect of head activator on nerve-cell determination and differentiation in hydra.

In hydra, head activator (HA) acts as positive signal for nerve-cell determination and differentiation. For both events, HA uses cAMP as the second messenger. Evidence is presented that the cAMP agonist, Sp-cAMPS, is able to mimick the effect of HA on nerve-cell determination and differentiation and that it is blocked by the antagonist Rp-cAMP. An adenylyl cyclase associated protein, CAP, appears to be involved as mediator for transducing the signal from the transmembrane HA receptor to the cAMP system. A cDNA coding for hydra CAP was isolated from the multiheaded mutant of Chlorohydra viridissima. The hydra CAP shows extensive homology with the yeast and, more so, mammalian CAPs. In hydra, CAP mRNA is expressed abundantly in interstitial and epithelial cells. The effect of HA, but not of cAMP, on nerve-cell differentiation was inhibited by pretreatment of hydra with a cap antisense oligonucleotide, suggesting a role for CAP as mediator in the signal transduction cascade between HA and cAMP.

Amino Acid Sequence↗
PubMed2006

Degeneration after sexual differentiation in hydra and its relevance to the evolution of aging.

Aging occurs in most multicellular animals, yet some primitive animals do not show any sign of aging. This raises the following question: How have metazoans acquired the trait of aging in the course of evolution? Comparative studies of various species have provided a clue to this question by showing that sexually reproducing organisms predominantly undergo aging. The evolutionary theory "pleiotropy" also postulates aging as a price for facilitating the reproduction in the early life stage of an organism. For investigating the association between sexual reproduction and aging, a sexual phase-inducible organism in a laboratory would be suitable. One of such organisms is hydra, a genus of Cnidaria. Asexual hydra has been considered to be immortal, but there is the possibility that hydra undergoes aging after sexual reproduction. To search for signs of aging in hydra, we studied sexually differentiated Hydra oligactis at the individual and cellular levels. As a result, we found a significant decline in the capacities for food capture, contractile movements, and reproduction. More importantly, we discovered an exponential increase in the mortality rate of the population. These observations suggest that the degenerative process in H. oligactis represents the aging process. Furthermore, we found that the number of germ cells increased, whereas the number of somatic cells concomitantly decreased. The observed change of the cell composition is thus consistent with the "pleiotropy" theory of aging.

Animals↗
PubMed1998

Phospholipase C cDNAs from sponge and hydra: antiquity of genes involved in the inositol phospholipid signaling pathway.

To know whether or not the set of genes involved in the inositol phospholipid signaling pathway already existed in the early evolution of animals, we carried out cloning of cDNAs encoding phospholipase Cs (PLCs) from Ephydatia fluviatilis (freshwater sponge) and Hydra magnipapillata strain 105 (hydra). We isolated two PLC cDNAs, PLC-betaS and PLC-gammaS, from sponge and three cDNAs, PLC-betaH1, PLC-betaH2, and PLC-deltaH, from hydra. From the domain organization and the divergence pattern in the PLC family tree, the sponge PLC-betaS and PLC-gammaS and the hydra PLC-deltaH are possibly homologous to the vertebrate PLC-beta, PLC-gamma and PLC-delta subtypes, respectively. A detailed phylogenetic analysis suggests that the hydra PLC-betaH1 and PLC-betaH2 are homologs of the vertebrate PLC-beta1/2/3/Drosophila PLC21 and the vertebrate PLC-beta4/Drosophila norpA, respectively. A phylogenetic analysis of the PLC family and the protein kinase C (PKC) family, together with that of the G protein alpha subunit (Galpha) family, revealed that the origin of the set of genes G(alpha)q, PLC, PKC involved in the inositol phospholipid signaling pathway is very old, going back to dates before the parazoan-eumetazoan split, the earliest branching among extant animal phyla.

Animals↗
PubMed1999

Acute toxicity assessment of Polish (waste) water with a microplate-based Hydra attenuata assay: a comparison with the Microtox test.

The use of Hydra attenuata in acute toxicity assessment is a potentially useful tool in (waste) water biomonitoring. The purpose of this study was to compare the sensitivity of H. attenuata with the extensively used Microtox test on 14 (waste) water samples from the Kraków region (South Poland). To this end, specific morphological changes displayed by the freshwater cnidarian Hydra attenuata (lethal LC50s and sublethal EC50s effects) and bioluminescence of the marine bacteria Vibrio fisheri (Microtox) were compared. Clearly, the Hydra assay was the more sensitive indicator of toxicity. No relationship was found among Hydra toxicological responses and water levels of As, Cd, Co, Cu, Pb and Zn. However, it appeared that toxicity to Hydra might be due to ammonia levels. Additional studies to better circumscribe the tolerance of H. attenuata to 'natural' water characteristics are needed.

Animals↗
PubMed1999

Identification of caspases and apoptosis in the simple metazoan Hydra.

Apoptosis is a normal process by which cells die and are eliminated from tissue by phagocytosis [1]. It is involved in regulating cell numbers in adult tissues and in eliminating 'excess' cells during embryogenesis and development. Apoptosis is mediated by activation of caspases, which then cleave a variety of cellular substrates and thereby cause the characteristic morphology of apoptotic cells (rounded cells, condensed chromatin, susceptibility to phagocytosis) [2]. Although apoptosis has been well documented in nematodes, insects and mammals, it is not yet clear how early in evolution apoptosis or its component enzymes arose. In the simple metazoan Hydra vulgaris, cell death regulates cell numbers [3] [4] [5]. In starved animals, for example, epithelial cell proliferation continues at a nearly normal rate although the tissue does not increase in size; the excess cells produced are eliminated by phagocytosis. Cell death can also be induced in wild-type hydra by treatment with colchicine [6] or in a mutant strain (sf-1) by temperature shock [7]. Here, we show that cell death in hydra is morphologically indistinguishable from apoptosis in higher animals, that hydra polyps express two genes with strong homology to members of the caspase 3 family, and that caspase-3-specific enzyme activity accompanies apoptosis in hydra. The occurrence of apoptosis and caspases in a member of the ancient metazoan phylum Cnidaria supports the idea that the invention of apoptosis was an essential feature of the evolution of multicellular animals.

Animals↗
PubMed1998

In situ potentiation of the glutathione-binding protein for the tentacle ball formation by a protease and efficient ingestion of prey in hydra.

Within minutes, brief treatment with trypsin potentiated tentacle ball formation in Hydra japonica, a new behavioral response to reduced glutathione. With the potentiation of this behavioral response, new glutathione-binding proteins were immediately detected after the trypsin treatment of live Hydra, indicating that trypsin activated the glutathione-binding protein in situ. Fixed brine shrimp (Artemia francisca) were more efficiently ingested in the presence of trypsin and S-methylglutathione (GSM) than in the presence of GSM alone, suggesting a biological role of this behavioral potentiation by trypsin in the feeding chain of Hydra. Ingestion of live A. francisca was significantly reduced in the presence of soybean trypsin inhibitor, suggesting that a protease, possibly released from the wounded prey, plays a role in the feeding in vivo. As for Hydra swallowing its captured prey, a small hydra head piece was isolated and measured as it crept along a thin nylon line; advancement of the head was the same in the presence of both GSM alone, and in that of GSM and trypsin together. Together, these results indicate that the chemoreceptor potentiated in situ by a trypsin-like protease specifically evokes tentacle ball formation resulting in an efficient transfer of prey on the tentacle to the mouth.

Animals↗
PubMed1980

Epithelial cells in nerve-free hydra produce morphogenetic substances.

It has been shown that hydra, consisting almost exclusively of epithelial cells, have normal morphogenetic properties. This means that they are able to reproduce asexually by budding and to regenerate head or foot with conservation of the original polarity. Such hydra have no interstitial cells and they lack nerves and nematocytes as differentiation produces of the interstitital stem cell. Consequently, they are unable to catch or ingest food and would not survive in ntaure. However, stocks can be maintained in the laboratory, if food is injected into the gastric cavity. The fact that nerve-free hydra would grow and regenerate normally raised some doubts as to the role of the nervous system in the control of differentiation and development in hydra. In particular, the significance of morphogenetic substances, such as the head activator, which is normally produced by nerve cells, seemed to need reassessment. We now show that epithelial cells, seemed toneed reassessment. We now show that epithelial cells in hydra have the potential to produce head activator and other morphogenetic factors, but that this property is repressed in the presence of nerves.

Animals↗
PubMed1998

The lack of a stress response in Hydra oligactis is due to reduced hsp70 mRNA stability.

Synthesis and degradation of hsp70 mRNA was examined and compared in Hydra species living in different habitats and showing different heat-shock response. Hydra oligactis is restricted to habitats of low temperature and relatively stable pH. We have shown previously that this species is unable to acquire thermotolerance [Bosch, T., Krylow, S., Bode, H. & Steele, R. (1988) Proc. Natl. Acad. Sci. USA 85, 7927-7931] and synthesizes significantly less heat-shock protein and hsp70 mRNA [Gellner, K., Praetzel, G. & Bosch, T. C. G. (1992) Eur J. Biochem. 210, 683-691] in response to stress than related species, such as Hydra bulgaris or Hydra magnipapillata, which are adapted to habitats of wide temperature range and variable water quality. To examine the mechanisms responsible for the differential heat-shock responses in these species, a construct containing H. magnipapillata hsp70 regulatory sequences fused to firefly luciferase was introduced into H. oligactis and H. magnipapillata polyps, and expression of luciferase examined. The results showed that luciferase can be expressed equally well in a heat-inducible manner in both species, suggesting that H. oligactis heat-shock factor can interact with H. magnipapillata heat-shock elements. Northern blots of alpha-amanitin-treated polyps demonstrated that the half-life of hsp70 mRNA in heat-shocked H. oligactis is drastically shorter than in H. magnipapillata. Thus, differences in hsp70 mRNA stability appear to be responsible for the habitat-correlated differences in the stress response in Hydra species.

Animals↗
PubMed1997

Evolution of paired domains: isolation and sequencing of jellyfish and hydra Pax genes related to Pax-5 and Pax-6.

Pax proteins are a family of transcription factors with a highly conserved paired domain; many members also contain a paired-type homeodomain and/or an octapeptide. Nine mammalian Pax genes are known and classified into four subgroups: Pax-1/9, Pax-2/5/8, Pax-3/7, and Pax-4/6. Most of these genes are involved in nervous system development. In particular, Pax-6 is a key regulator that controls eye development in vertebrates and Drosophila. Although the Pax-4/6 subgroup seems to be more closely related to Pax-2/5/8 than to Pax-3/7 or Pax-1/9, its evolutionary origin is unknown. We therefore searched for a Pax-6 homolog and related genes in Cnidaria, which is the lowest phylum of animals that possess a nervous system and eyes. A sea nettle (a jellyfish) genomic library was constructed and two pax genes (Pax-A and -B) were isolated and partially sequenced. Surprisingly, unlike most known Pax genes, the paired box in these two genes contains no intron. In addition, the complete cDNA sequences of hydra Pax-A and -B were obtained. Hydra Pax-B contains both the homeodomain and the octapeptide, whereas hydra Pax-A contains neither. DNA binding assays showed that sea nettle Pax-A and -B and hydra Pax-A paired domains bound to a Pax-5/6 site and a Pax-5 site, although hydra Pax-B paired domain bound neither. An alignment of all available paired domain sequences revealed two highly conserved regions, which cover the DNA binding contact positions. Phylogenetic analysis showed that Pax-A and especially Pax-B were more closely related to Pax-2/5/8 and Pax-4/6 than to Pax-1/9 or Pax-3/7 and that the Pax genes can be classified into two supergroups: Pax-A/Pax-B/Pax-2/5/8/4/6 and Pax-1/9/3/7. From this analysis and the gene structure, we propose that modern Pax-4/6 and Pax-2/5/8 genes evolved from an ancestral gene similar to cnidarian Pax-B, having both the homeodomain and the octapeptide.

Amino Acid Sequence↗
PubMed2007

Nerve ring of the hypostome in hydra: is it an origin of the central nervous system of bilaterian animals?

A hypothesis, 'the nerve ring in hydra shares a common origin with the central nervous system in bilaterian animals', is discussed in this review. The nerve ring of hydra is a ring of neurons whose neurites make a bundle running circumferentially around the hypostome just above the tentacle zone. This nervous structure has unique features in the hydra nervous system. It shows a tight association of neurons in contrast to the diffuse nerve net seen in other regions. It shows static developmental characters in contrast to the dynamic features of hydra nerve net present in other regions. Moreover, its structure and location are similar to the central nervous system (CNS) of other animals without a complex CNS such as nematodes and starfishes. Functions of the hydra nerve ring are also studied to test the hypothesis. The identified function is a crumpling of the tentacles, corresponding to the function of the inner nerve ring of hydrozoan jellyfish. The jellyfish nerve ring is considered to be a primitive central nervous system of radiates. Considering all the information available, the hypothesis is highly possible.

Animals↗
PubMed1995

The cAMP response element binding protein is involved in hydra regeneration.

Hydra provides an interesting developmental model system where pattern formation processes are easily accessible to experimentation during regeneration. Previous studies have shown that the neuropeptide head activator affects cellular growth and head-specific cellular differentiation during head regeneration and budding. In order to investigate the signal transduction pathway and the regulatory genes involved in these processes, we measured cAMP levels after head activator treatment and found that head activator leads to an increase in cAMP levels at concentrations where effects on nerve cell determination and differentiation are observed (10(-11) to 10(-9) M). Moreover, exposure of intact hydra to a permeable form of cAMP stimulates nerve-cell differentiation and thus mimicks the effect of endogenous head activator. Band-shift assays were performed to detect changes in hydra nuclear protein binding activity during regeneration or after head activator treatment. We found that the cAMP response element (CRE) promotes a specific and strong DNA-binding activity which is dramatically enhanced and modified during early regeneration or after HA treatment. We also identified a surprisingly highly conserved hydra gene encoding the cAMP Response Element Binding protein, which is involved in this CRE-binding activity. Initiation of regeneration upon wounding provokes an endogenous release of HA which leads to the final differentiation of determined nerve cells. We propose that the nerve-cell differentiation observed within the first 4-8 hours of regeneration relies on the agonist effect of head activator on the cAMP pathway, which would in turn modulate the CRE-binding activity of the hydra CREB protein and thus regulate the transcriptional activity of genes involved in regeneration processes.

Activating Transcription Factor 2↗
PubMed2000

Identification and characterization of hydra metalloproteinase 2 (HMP2): a meprin-like astacin metalloproteinase that functions in foot morphogenesis.

Several members of the newly emerging astacin metalloproteinase family have been shown to function in a variety of biological events, including cell differentiation and morphogenesis during both embryonic development and adult tissue differentiation. We have characterized a new astacin proteinase, hydra metalloproteinase 2 (HMP2) from the Cnidarian, Hydra vulgaris. HMP2 is translated from a single mRNA of 1.7 kb that contains a 1488 bp open reading frame encoding a putative protein product of 496 amino acids. The overall structure of HMP2 most closely resembles that of meprins, a subgroup of astacin metalloproteinases. The presence of a transient signal peptide and a putative prosequence indicates that HMP2 is a secreted protein that requires post-translational processing. The mature HMP2 starts with an astacin proteinase domain that contains a zinc binding motif characteristic of the astacin family. Its COOH terminus is composed of two potential protein-protein interaction domains: an "MAM" domain (named after meprins, A-5 protein and receptor protein tyrosine phosphatase mu) that is only present in meprin-like astacin proteinases; and a unique C-terminal domain (TH domain) that is also present in another hydra metalloproteinase, HMP1, in Podocoryne metalloproteinase 1 (PMP1) of jellyfish and in toxins of sea anemone. The spatial expression pattern of HMP2 was determined by both mRNA whole-mount in situ hybridization and immunofluorescence studies. Both morphological techniques indicated that HMP2 is expressed only by the cells in the endodermal layer of the body column of hydra. While the highest level of HMP2 mRNA expression was observed at the junction between the body column and the foot process, immunofluorescence studies indicated that HMP2 protein was present as far apically as the base of the tentacles. In situ analysis also indicated expression of HMP2 during regeneration of the foot process. To test whether the higher levels of HMP2 mRNA expression at the basal pole related to processes underlying foot morphogenesis, antisense studies were conducted. Using a specialized technique named localized electroporation (LEP), antisense constructs to HMP2 were locally introduced into the endodermal layer of cells at the basal pole of polyps and foot regeneration was initiated and monitored. Treatment with antisense to HMP2 inhibited foot regeneration as compared to mismatch and sense controls. These functional studies in combination with the fact that HMP2 protein was expressed not only at the junction between the body column and the foot process, but also as far apically as the base of the tentacles, suggest that this meprin-class metalloproteinase may be multifunctional in hydra.

Amino Acid Sequence↗
PubMed1991

A 33 kDa protein with sequence homology to the 'laminin binding protein' is associated with the cytoskeleton in hydra and in mammalian cells.

In hydra and in mammalian cells the monoclonal antibody V recognises an epitope which colocalises with cytoskeletal structures. Using this antibody for expression screening, a cDNA clone (955 bp) was isolated from hydra, which covers an open reading frame for a protein of 294 amino acids with a calculated molecular mass of 32.8 kDa. Northern blot analysis of hydra RNA resulted in a single mRNA species of 1.2 kb, and primer extension experiments proved this to be the full length message. 218 residues at the amino terminus of the hydra protein show extensive homology (73.5%) to a human protein designated 'laminin binding protein'. The carboxyl-terminal 76 amino acids possess no significant similarity (20%). The monoclonal antibody V, which recognises an epitope in this carboxyl-terminal part, reacts in Western blots, both in hydra and in mammalian cells, with a protein of 33 kDa and not with the 45 kDa 'laminin binding protein'. The 33 kDa protein is not extracellular or transmembrane, but has a strictly intracellular location as indicated by its amino acid sequence and by immunocytochemical and cell fractionation studies. In non-dividing mammalian cells the 33 kDa protein colocalises with filamentous structures; in dividing cells it dissociates from it and concentrates centrally. Presence of the SPLR-sequence, which is the consensus phosphorylation motif for the p34cdc2 kinase, links this 33 kDa protein to events occurring during the cell cycle.

3T3 Cells↗
PubMed1996

Hydra regeneration from recombined ectodermal and endodermal tissue. I. Epibolic ectodermal spreading is driven by cell intercalation.

Cell-cell interaction and cell rearrangement were examined in the process of epithelial sheet formation during regeneration from hydra cell aggregates. The ectodermal and endodermal epithelial cell layers of Hydra magnipapillata were separated by procaine treatment. Each of the separated layers was then dissociated into single cells and reaggregated to produce ectodermal or endodermal cell aggregates. When the two aggregate types were recombined, a firm adhesion was quickly established between them. This was followed by a vigorous spreading of the ectodermal epithelial cells as a thin layer over the endoderm in a manner similar to the 'epiboly' in some developing embryos. Cell movement in this spreading process was examined using fluorescent dyestaining. It revealed that cells initially located in the inside of the aggregate migrated to intercalate themselves among the cells originally present in the contact surface. This radial cell intercalation took place continuously in the contact surface of both the ectodermal and endodermal aggregates, and produced a rapid growth of the contact surface, eventually leading to complete envelopment of the entire endoderm by the ectoderm. The resulting structure was a small sphere having a two-layered epithelial organization as in normal hydra. This sphere regenerated into a complete hydra a few days later. A tryptic extract of hydra membrane fraction specifically inhibited the ectodermal spreading over the endoderm, but not the initial adhesion or the later regeneration processes. These observations suggest that radial cell intercalation at the contact surface plays a crucial role in producing ectodermal spreading and establishing epithelial sheet organization in the recombined aggregates. The intercalation is presumably activated by a signal exchange through the contact surface. The inhibitory effect of the membrane extract suggests that it contains a factor that is involved in some way in this signaling mechanism.

Animals↗
PubMed1977

Tissue economics of hydra: regulation of cell cycle, animal size and development by controlled feeding rates.

Epithelial cell production and epithelial cell loss in 6 different size classes of Hydra attenuata were examined to understand the relationships between growth and morphogenesis. The sizes of adult hydra, the sizes of their buds, and their budding rates are all nearly proportional to the amount of food the hydra eat. Hydra fed at high rates (4-25 Artemia nauplii per day) all have the same epithelial cell cycle time (about 4 days). Budding accounts for most of their cell loss. Hydra fed 4-12 Artemia per day maintain a steady state condition in which tissue loss balances tissue growth. Animals fed 25 Artemia per day are not in a steady state growth condition and change in size. At the lowest feeding rates (0-1 Artemia per day), the epithelial cell cycle time is lengthened to about 16 days. Cell loss from the tentacles accounts for most of the cell loss, and this loss is not completely balanced by growth. As a consequence these animals cease budding and shrink in size.

Animals↗
PubMed2002

Epithelial interactions in Hydra: apoptosis in interspecies grafts is induced by detachment from the extracellular matrix.

Apoptosis plays an important role in immunity and is widely used to eliminate foreign or infected cells. Cnidaria are the most basal eumetazoans and have no specialised immune cells, but some colonial cnidarians possess a genetic system to discriminate between self and non-self. By grafting epithelia of different species we have previously shown that the freshwater polyp Hydra eliminates non-self cells by phagocytosis. Here we have investigated whether apoptosis is involved in the histocompatibility reactions. We studied epithelial interactions between Hydra vulgaris and Hydra oligactis and show that a large number of apoptotic cells accumulate in the contact region of interspecies grafts. Histological analysis of the graft site revealed that displacement of the endodermal layer of Hydra vulgaris by endoderm from Hydra oligactis coincided with impaired cell-cell and cell-matrix contacts. We therefore suggest that in interspecies grafts, apoptosis is induced by the detachment of epithelial cells from the extracellular matrix (anoikis) and not by a discriminative allorecognition system.

Animals↗
PubMed1986

Bud induction in decapitated Hydra attenuata by 5-azacytidine: a morphological study.

The effect of 5-azacytidine (5-azaCR) on head regeneration and budding in hydra are reported. Hydra attenuata were exposed to various doses of 5-azaCR for 48 h and then decapitated and cultured. Head regeneration and bud formation were observed for 12 days after decapitation. Untreated control hydra regenerated heads within 7 to 8 days of decapitation with a budding index of 0.2. Buds invariably arose in the normal budding zone (below the gastric region). In the group treated with 0.8 mM-5-azaCR, 9 days after decapitation head regeneration was seen in only 13% of animals, and an average of two buds per hydra were formed, most of which were in the vicinity of the distal end. Induction of budding was also seen in the animals that regenerated heads. In animals exposed to 1 mM-5-azaCR three main types of responses were observed 9 days after decapitation. 44% of the animals regenerated normal heads; about half of them developed at least one bud and these buds originated in the budding zone. 17.5% of the animals developed abnormal, long hypostome-like structures with single or bifurcated tentacles at their tips. There were at least two buds per animal and they were invariably at abnormal sites. 32% of the animals failed to regenerate heads, although they developed two buds. 87% of these buds originated in abnormal sites of the body column and a large number (72%) did not detach even by the 12th day after decapitation. Both 5 and 10 mM of 5-azaCR were toxic to the animals; the survivors formed large globe-shaped heads. Bud induction was seen in 60% and 28% of animals in the 5 and 10 mM groups, respectively. These observations demonstrate that 5-azaCR induces bud formation in hydra at doses that inhibit head regeneration. This bud induction might be due to a specific expression of gene products responsible for bud formation.

Animals↗