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At least 19 recordsLinked to original sources
PubMed1979

Genetic analysis of developmental mechanisms in hydra. VI. Cellular composition of chimera hydra.

The homeostatic mechanisms that maintain constant cellular ratios in hydra tissue were studied using mutant and chimeric hydra strains. Mutants having abnormal cellular compositions are isolated through sexual inbreeding of wild hydra, as described in previous papers of this series. Chimeric hydra are produced by making use of a strain (nf-I) which lacks interstitial cells, nerve cells and nematocytes in its tissue. Reintroduction of interstitial cells from other strains (both normal and mutant) into nf-I leads to creation of chimeric strains having epithelial cell lineages from one strain (nf-I) and interstitial cell lineages from others. Analyses and comparisons of the cellular compositions of all these strains revealed that the numbers of nerve or interstitial cells in the chimeras were very similar to (statistically significantly correlated with) those in their interstitial cell donors. Since chimeras and their interstitial cell donors share the same interstitial cell lineages, this suggests that interstitial cells or their derivatives (nerves and nematocytes) play major roles in determining the nerve and interstitial cell levels in the hydra tissue. It is suggested that some form of homeostatic feedback mechanisms are probably involved in regulating the levels of these cell types.

Animals↗
PubMed1975

Positional information and pattern regulation in hydra: enzyme profiles.

Certain key enzymes of alternative pathways of glucose metabolism, of amino acid metabolism and of redox systems have been measured in hydra and this profile compared with mammalian differentiated tissues with a view to locating pathways of specific importance in hydra. There was a marked constant proportionality in the major part of the enzymes investigated, the profile suggested a metabolic pattern geared to utilization of amino acids as a carbon source for biosynthesis and energy production and to the production and conservation of pyruvate. The importance of conversion to ionized forms was noted. The most notable specific proportion changes were the exceptionally low lactate dehydrogenase, malic enzyme and the relatively high citrate synthase. The proximal-distal gradients in hydra were examined and these gradients suggested a switch to a more anaerobic type of metabolism and an elevation of the pentose phosphate pathway as the basal region was approached. Measurements of the formation of 14CO2 from specifically labelled glucose provided additional evidence for the functional activity and polarity of the pentose phosphate pathway in hydra. The effect of oligomycin, which can reverse polarity in hydra, had a significant effect on gradients of enzymes eliminating all except that observed for G6P dehydrogenase. The profile suggested a movement towards a more anaerobic type of metabolism, in keeping with the known biochemical action of this inhibitor. It is suggested that redox states and/or phosphorylation states may be featured in the positional information of cells in hydra.

Alanine Transaminase↗
PubMed1979

Colchicine, nocodazole and trifluralin: different effects of microtubule polymerization inhibitors on the uptake and migration of endosymbiotic algae in Hydra viridis.

Hydra viridis (= Chlorohydra viridissima) the freshwater coelenterate, is symbiotic. Each individual animal harbours Chlorella sp. in its endodermal cells. The symbiosis may be disestablished, the partners grown independently, and then re-established experimentally. The most effective method to produce alga-free hydra was developed by Pardy (1976). In this study algae from homogenized H. viridis, English (= European) strain, were either used directly or first grown in pure culture and then injected to re-establish the symbiosis. The uptake of algae grown in culture was compared with that of algae released directly from fresh hydra. Fewer cultured algae were taken up. Four strains were tested: English, Carolina, Frome, and Jubilee. Each takes up a characteristic number of Chlorella when injected in excess. Colchicine, nocodazole and trifluralin, were tested for their effects on either uptake or migration. Colchicine had no effect. Nocodazole and trifluralin reduced both the number of algae taken up and the rate at which they were transported to the distal region of the hydra digestive cell. The effects of these drugs tended to be proportional to the concentration between 10(-8) and 10(-5) M.

Animals↗
PubMed1978

Regulation of the self-renewal probability in Hydra stem cell clones.

Hydra interstitial stem cells continuously give rise to daughter stem cells as well as precursors for nerve and nematocyte differentiation. Growth of the stem cell population is controlled by the self-renewal probability (Ps): Ps is the fraction of stem cell daughters that remain stem cells in each generation. We have determined Ps for Hydra interstitial stem cells by using a novel technique based on the cell conposition of clones. Stem cell clones were grown in aggregates of nitrogen mustard-inactivated Hydra tissue. They contain several hundred cells after 14 days of growth, including stem cells, differentiating nematocytes, and differentiating nerve cells. Clone size, size variability, and the ratio of differentiating cells to stem cells are sensitive measures of Ps. We have prepared standard curves relating these parameters to Ps, using computer simulations of clone growth. Comparisoon of the experimentally observed parameter of clones to these curves indicates that Ps decreases from 0.8 in 5- to 6-day clones to 0.6 in 10- to 12-day clones. The decrease in Ps coincides with the increase in clone size and suggest that Ps may be regulated by the density of stem cells in clones. Such a mechanism could be responsible for the observed homeostasis of stem cell populations in vivo.

Cell Differentiation↗
PubMed1979

Production of nerveless Hydra attenuata by hydroxyurea treatments.

Hydroxyurea was used to produce hydra with varying nerve cell densities including a new type of nerveless animal. Hydra attenuata were treated with 10(-2) M hydroxyurea. By 23 days after treatment, 2 populations of animals are in culture. Both have a decrease in nerve cells. The first is a normal-coloured feeding animal (HU-R) and is recovering while the second is a pale non-feeding animal (HU-P). HU-P animals resemble nerveless animals in their lack of behavioural responses but they contain about 2% nerve cells. Upon hand feeding, some HU-P animals will recover but most will produce nerveless buds. Neverless hydra produced by hydroxyurea resemble nerveless animals produced by other techniques, in their behavioural, morphological and developmental properties. Normal animals, as cultured and in regeneration experiments, show a compact tentacle number distribution pattern with a small variance. Nerveless animals show broad tentacle distribution patterns with increased means and variances. It is suggested that a normal tentacle number regulatory mechanism is lacking or diminished in nerveless animals. This defect is correlated with hypostomal circumference and with nerve cells.

Animals↗
PubMed1979

Stem cell growth and differentiation in Hydra attenuata. I. Regulation of the self-renewal probability in multiclone aggregates.

Interstitial stem cells in Hydra are rapidly proliferating multipotent stem cells which continuously give rise to precursors for nerve and nematocyte differentiation. Growth of the stem cell population is controlled by the cell cycle time of the stem cells and the self-renewal probability, Ps (the fraction of stem cells in each generation which divide to yield more stem cells). In normal Hydra the stem cell generation time is 24 h and Ps = 0.6; under these conditions the stem cell population doubles in 3.5 days. In the present experiments we have systematically investigated the dependence of Ps on stem cell density. We culture stem cells in a feeder layer system consisting of aggregates of nitrogen-mustard (NM)-inactivated Hydra cells. In this system stem cell density can be varied over a wide range by changing the number of clone-forming units (CFU) added to the aggregates. We have measured the growth rate of the stem cell population and the cell cycle of stem cells in NM aggregates after 4--7 days of culture. From these data we calculate the value of Ps. The results indicate that the growth rate decreases 4-fold as the number of CFU seeded per aggregate increases from 10 to 400. Under these same conditions the cell cycle remains constant. The values of Ps calculated from these results indicate the Ps decreases from 0.75 in aggregates seeded with 10--30 CFU to 0.55 in aggregates seeded with 200--400 CFU. These results support a model in which Ps is controlled by negative feedback from neighbouring stem cells. In addition, our experiments indicate that Ps decreases during the growth of stem cell clones. When only a few stem cells are seeded in aggregates, they give rise to isolated clones distributed throughout the aggregate. Ps decreases markedly within such clones as they grow in size presumably due to increasing stem cell content of the clones. Since Ps in such isolated clones declines with growth, we infer that the local stem cell concentration is what controls Ps and that the spatial range of the negative feedback signal is short compared to the dimensions of NM aggregates.

Animals↗
PubMed2016

Loss of neurogenesis in Hydra leads to compensatory regulation of neurogenic and neurotransmission genes in epithelial cells.

Hydra continuously differentiates a sophisticated nervous system made of mechanosensory cells (nematocytes) and sensory-motor and ganglionic neurons from interstitial stem cells. However, this dynamic adult neurogenesis is dispensable for morphogenesis. Indeed animals depleted of their interstitial stem cells and interstitial progenitors lose their active behaviours but maintain their developmental fitness, and regenerate and bud when force-fed. To characterize the impact of the loss of neurogenesis in Hydra, we first performed transcriptomic profiling at five positions along the body axis. We found neurogenic genes predominantly expressed along the central body column, which contains stem cells and progenitors, and neurotransmission genes predominantly expressed at the extremities, where the nervous system is dense. Next, we performed transcriptomics on animals depleted of their interstitial cells by hydroxyurea, colchicine or heat-shock treatment. By crossing these results with cell-type-specific transcriptomics, we identified epithelial genes up-regulated upon loss of neurogenesis: transcription factors (Dlx, Dlx1, DMBX1/Manacle, Ets1, Gli3, KLF11, LMX1A, ZNF436, Shox1), epitheliopeptides (Arminins, PW peptide), neurosignalling components (CAMK1D, DDCl2, Inx1), ligand-ion channel receptors (CHRNA1, NaC7), G-Protein Coupled Receptors and FMRFRL. Hence epitheliomuscular cells seemingly enhance their sensing ability when neurogenesis is compromised. This unsuspected plasticity might reflect the extended multifunctionality of epithelial-like cells in early eumetazoan evolution.

Animals↗
PubMed1976

The morphology of green hydra endosymbionts as influenced by host strain and host environment.

The ultrastructure of Chlorella-like algal endosymbionts from the Florida and English strains of green hydra was compared under different host feeding and photoperiodic regimes. Under standard conditions (host fed daily, 12-h photoperiod) the algae from the 2 strains exhibited considerable differences. The English symbionts had a pyrenoid, compact chloroplast membranes and vesiculated polyphosphate bodies. By comparison, Florida symbionts lacked a pyrenoid, had chloroplasts with less compact membranes and exhibited spherical polyphosphate bodies. When maintained in the dark, algae from English hydra lost their pyrenoids, showed great compaction of the chloroplast and developed large, shield-shaped, electron-dense bodies. In contrast, algae from Florida hosts did not exhibit gross ultrastructural modification. Reciprocal cross-transfers of symbionts were made by placing Florida algae in English aposymbiotic (algal-free) hosts and vice versa. After residence in Florida hosts, English symbionts appeared to undergo ultrastructural modifications resulting in a morphology indistinguishable from the native Florida symbionts. Florida algae showed no modifications resulting from residence in English hosts. It thus appears that the English symbiont has great morphological plasticity, as its structure is greatly modified depending upon the host in which it resides and the conditions under which the host is maintained. The results of these studies are discussed and compared with published accounts of free-living Chlorella and with reports dealing with other Chlorella symbionts.

Chlorophyta↗
PubMed1975

Investigation of oogenesis in hydra. Communication I. Ultrastructure of interstitial cells at early stages of their transformation into oocytes.

We studied ultrastructure of the interstitial cells (i-cells) at successive stages of their transformation into oocytes. The i-cells that form the epidermal thickening in the genital zone of the animal start to proliferate in hydras approaching sexual reproduction. Undifferentiated i-cells are characterized by a high density of diffusely disposed ribosomes and almost total absence of membranous structures. Femal sex cells starting to undergo differentiation are characterized by the appearance of aggregations of finely filamentous material in the cytoplasm. This material is of nuclear origin and is often surrounded by mitochondria. Dimensions of the i-cells increase rapidly, and single elongated cisterns of the granular endoplasmic reticulum appear in them. These cisterns run parallel to the cell surface and give rise to minute smooth-surfaced vesicles, whose membranes fuse wtih the cytoplasmic membrane in such a way that their contents are emptied into the intercellular space. Different sectors of the cell are marked by the appearance of grouped cisterns of the granular endoplasmic reticulum. Such cisterns give rise to smooth-surfaced vesicles, which form numerous Golgi complexes. The number of mitochondria increases simultaneously; aggregations of lipid drops with glycogen granules among them start to appear; and escape of nuclear material into the cytplasm intensifies. Such cells at the stage of cytoplasmic growth and accumulation of organelles and yolk inclusions. In connection with this, it is suggested that the term "oogonium" (which is used to designate such cells in the literature) be retained only for proliferating i-cells containing finely filamentous nuclear material in the cytoplasm in the genital zone of the hydra.

Animals↗
PubMed1976

The general occurrence of intramitochondrial crystals in Hydra cells.

Intramitochondrial crystals are found in normal Hydra as well as in animals undergoing various conditions (budding, regenerating, eserine-treated, and sexual). They appear in all regions of the animal, but seem to be more prevalent at or near the extremities: hypostome, tentacles and basal disk. They are found in all of the seven basic cell types: interstitial, cnidocyte, nerve, epithelio-muscular, gland, mucous and digestive cells. The chemical nature of the intramitochondrial crystals is unknown and their significance remains speculative.

Animals↗
PubMed1979

The fine structure of the hypostome and mouth of hydra. I. Scanning electron microscopy.

The hypostome and mouth of fresh-water Hydra were examined by scanning electron microscopy. The external surface of the hypostome possesses cnidocils, possibly sensory hairs, and small spiny protrusions surrounding the mouth; the internal surface has cylindrical microvilli, free flagella and adherent flagella. The adherent flagella are most numerous close to the mouth where they cause the cell surface to appear smooth when viewed at low magnifications. Free flagella and leaf-like microvilli increase in prominence towards the tentacles and enter on proper. The edge of the mouth has an abrupt boundary marking the apposition of epidermal and gastrodermal cells. A transitional groove occurs at the boundary and the cells underlying the groove are smaller than those on other regions of the hypostome. The transition groove may represent a site of cell loss in normal cell turnover. Some of the small underlying cells may represent nervous elements involved in regulating hypostome activity during the feeding reation.

Animals↗
PubMed1979

The fine structure of the hypostome and mouth of hydra. II. Transmission electron microscopy.

The normal morphology of the hypostome and mouth of hydra were examined by transmission electron microscopy with conventional thin sections and freeze-fracture replicas. Myonemes of the hypostome are small in diameter, have gap and intermediate-type cell junctions within each epithelial layer and are associated with the opposite epithelial layer by transmesogleal processes and gap junctions. Nematocysts and sensory cells are aggregated in the circumoral region. The fine structure of adherent flagella arising from gastrodermal gland cells, and the transition region at the mouth between epidermis and gastrodermis are described in detail for the first time. The possible functional significance of the findings is discussed.

Animals↗
PubMed1978

A second sensory--motor--interneuron with neurosecretory granules in Hydra.

Using serial-sectioning techniques for conventional transmission and high-voltage electron microscopy, we characterized the ultrastructural features and synaptic contacts of the sensory cell in tentacles of Hydra. The sensory cell has an apical specialization characterized by a recessed cilium surrounded by three rodlike stereocilia. This ciliary--stereociliary complex constitutes the receptive or dendritic pole of the sensory cell. The dense filamentous cores of the stereocilia project proximally into a narrow circumciliary cytoplasmic region connected by septate junctions to marginal processes of an enveloping epitheliomuscular cell. The central cilium has a characteristic marginal flare midway along its length and a dense filamentous substructure at its base. Pairs of branched, striated rootlets extend from the axial centriole into a mitochondria-rich region of the cell. Pigment-like granules are present in the cytoplasm around the circumciliary space. The perikaryon is characterized by an elongate nucleus surrounded by a narrow rim of cytoplasm containing prominent Golgi complexes, assorted vacuoles and dense-cored vesicles, free ribosomes, short segments of rough endoplasmic reticulum, microtubules, glycogen particles, and lipid droplets. Generally, one or two thin, naked axons extend laterally from the perikaryon into the nerve net region above the myonemes of the large epitheliomuscular cells. Within the axons are found occasional aggregates of dense-cored vesicles and en passant synapses characterized by the presence of clear or dense-cored vesicles in contact with paramembranous densities and associated intracleft cross filaments. Using these ultrastructural criteria, we demonstrated for the first time that the granule-containing sensory cells have synaptic contacts with other neurons, nematocytes, and epitheliomuscular cells hence, we considered these cells to be sensory--motor--interneurons with neurosecretory granules. We hypothesize that this unique, apparently multifunctional neuron may be a modern representative of a primitive stem cell that give rise evolutionarily to the sensory cells, motor neurons, interneurons, and neurosecretory cells of higher animals.

Animals↗
PubMed1978

Interaction of glutathione analogues with Hydra attenuata gamma-glutamyltransferase.

gamma-Glutamyltransferase activity was studied in extracts of the cnidarian Hydra attenuata. The binding of gamma-glutamyl peptide analogues to the enzyme was studied by observing their effects on heat denaturation and their inhibition of p-nitroaniline release from gamma-glutamyl p-nitroanilide. Neither position-1 analogues, in which the gamma-glutamyl moiety was changed to a beta-aspartyl (beta-Asp-Abu-Gly) or an alpha-glutamyl (Glu-Abu-Gly) linkage, nor glutamate protected the enzyme against inactivation at 58 degrees C. GSH (reduced glutathione), gamma-Glu-Abu-Gly and gamma-Glu-Met on the other hand did prevent heat denaturation. GSH and analogues of GSH were competitive inhibitors of p-nitroaniline release, but those analogues in which glycine was replaced by 2-aminoisobutyrate, phenylalanine, leucine or tyrosine had Ki values that were approximately five times those of analogues with the cysteine residue replaced.

Amino Acids↗
PubMed1979

Stem cell growth and differentiation in Hydra attenuata. II. Regulation of nerve and nematocyte differentiation in multiclone aggregates.

The differentiation of nerve cells and nematocytes from interstitial stem cells in Hydra has been investigated under conditions of changing stem cell density. Interstitial stem cells were cultured in a feeder layer system consisting of aggregates of nitrogen mustard-inactivated tissue. The aggregates were seeded with varying numbers of stem cells from 10 to 400 per aggregate; between 4 and 7 days later the rates of nerve and nematocyte differentiation were measured. Nerve differentiation was scored by labelling the stem cell population with [3H]-thymidine and counting nests of 4 proliferating nematoblasts. In both cases the numbers of differentiating cells were normalized to the size of the stem cell population. The results indicate that the rate of nematocyte differentiation increases as the concentration of stem cells increases in aggregates; under the same conditions the rate of nerve differentiation remains essentially constant. To calculate the numbers of stem cells entering each pathway per generation, a computer was programmed to simulate the growth and differentiation of interstitial stem cells. Standard curves were prepared from the simulations relating the rates of nerve and nematocyte differentiation to the fraction of stem cells committed to each pathway per generation. The rates of nerve and nematocyte commitment were then estimated from the experimentally observed rates of differentiation using the standard curves. The results indicate that nerve commitment remains constant at about 0.13 stem cells per generation over a wide range of stem cell concentration. Nematocyte commitment, by comparison, increases from 0.15 to 0.21 stem cells per generation as stem cell concentration increases in aggregates. The fact that the ratio of nerve to nematocyte commitment changes under our conditions suggests that stem cell commitment is not a stochastic process but subject to control by environmental stimuli.

Animals↗
PubMed1979

Cell contact and positional communication in hydra.

Positional communication and the functional coupling of muscular reflexes were examined in grafted hydra. A head and distal gastric region inhibited head regeneration by a host sub-hypostome within 4--5 h of grafting. Functionally coupled pathways which indicated the presence of gap junctions also formed between graft and host during this time. It is suggested that gap junctions provide a channel for positional communication.

Animals↗