Studies on hydroid differentiation. 3. The replacement of hypostomal gland cells of Podocoryne carnea

Pulse labeling with tritiated thymidine has permitted the identification of a progenitor cell compartment at the base of the hypostomal glandular ridges of the marine hydroid Podocoryne carnea. This progenitor compartment produces cells which move out on the ridges differentiate as digestive gland cells, and are ejected into the gastro-vascular cavity. Spherous gland cells of the body appear not to be part of this circumscribed replacement system. There appears to be no proximal movement of gastrodermal cells in the body of this marine hydroid.     

Post-embryonic larval development and metamorphosis of the hydroidEudendrium racemosum (Cavolini) (Hydrozoa,Cnidaria)

The morphology and histology of the planula larva ofEudendrium racemosum (Cavolini) and its metamorphosis into the primary polyp are described from light microscopic observations. The planula hatches as a differentiated gastrula. During the lecithotrophic larval period, large ectodermal mucous cells, embedded between epitheliomuscular cells, secrete a sticky slime. Two granulated cell types occur in the ectoderm that are interpreted as secretory and sensorynervous cells, but might also be representatives of only one cell type with a multiple function. The entoderm consists of yolk-storing gastrodermal cells, digestive gland cells, interstitial cells, cnidoblasts, and premature cnidocytes. The larva starts metamorphosis by affixing its blunt aboral pole to a substratum. While the planula flattens down, the mucous cells penetrate the mesolamella and migrate through the entoderm into the gastral cavity where they are lysed. Subsequently, interstitial cells, cnidoblasts, and premature cnidocytes migrate in the opposite direction, i.e. from entoderm to ectoderm. Then, the polypoid body organization, comprising head (hydranth), stem and foot, all covered by peridermal secretion, becomes recognisable. An oral constriction divides the hypostomal portion of the gastral cavity from the stomachic portion. Within the hypostomal entoderm, cells containing secretory granules differentiate. Following growth and the multiplication of tentacles, the head periderm disappears. A ring of gland cells differentiates at the hydranth's base. The positioning of cnidae in the tentacle ectoderm, penetration of the mouth opening and the multiplication of digestive gland cells enable the polyp to change from lecithotrophic to planktotrophic nutrition.     

Ultrastructural observations on gastrodermal regeneration in the planarian Dugesia japonica (Turbellaria)

The earliest detectable change during regeneration of the gastrodermis in Dugesia japonica was an aggregation of regenerative cells underneath the gastrodermis remaining at the wound margin. The gastrodermal cells in experimental regenerates retained some of their original characters and presented no indication of cell dedifferentiation. The regenerative cells came into contact with the basal surface of gastrodermal cells, forming stratified cell layers. Differentiation of these cells into gastrodermal cells was initiated by the development of synthetic organelles within their cytoplasm. These differentiating cells gave rise to two different types of gastrodermal cells, namely phagocytic cells and sphere cells. In later stages, there was an apparent movement of differentiated gastrodermal cells towards the parenchyma.     

Regulation in the numbers of tentacles of aggregated hydra cells

Head formation was investigated during regeneration of dissociated and aggregated cells of Hydra magnipapillata. The surface area measured at the hollowing stage was found to be a useful quantity for characterizing the size of an aggregate. Four kinds of aggregates were examined, using tissue originating from (1) whole animals, (2) apical halves, (3) decapitated animals, and (4) decapitated animals allowed to regenerate for several hours before dissociation. For aggregate types (1), (2), and (4), not all the tentacles observed at an intermediate stage of the regeneration process were localized around hypostomes: the number of such body tentacles at the intermediate stage was comparable to that of the hypostomal tentacles and was approximately proportional to the surface area. These results and others suggest that the formation of body tentacles takes place independently of hypostome formation. However, for aggregate type (3), most of the tentacles appearing at the intermediate stage were hypostomal. The correlation between the surface area and the number of tentacles at the steady state apparently resulted from a regulation process by which body tentacles decreased and hypostomal tentacles increased. It is considered that the number of body tentacles appearing at an intermediate stage of regeneration would depend on the initial level of head-activation potential and that body tentacles are formed by the local fluctuation of head-activation potential.     

On the occurrence and significance of annulate lamellae in gastrodermal cells of regenerating planarians.

Cytoplasmic annulate lamellae have been observed to occur only in a subset of the gastrodermal cell population of regenerating planarians. They have not been found in the gastrodermal cells of intact, non-injured worms, nor in any other somatic cell type. These observations plus the presence of numerous chromatoid bodies in the same cells are consistent with the hypothesis that these cells are altering their state of differentiation and are preparing for division. It is further suggested that these cells are the precursors to the definitive somatic stem cells, the beta cells.     

Ultrastructure of the calcium-sequestering gastrodermal cell in the hydroid Hydractinia symbiolongicarpus (Cnidaria, Hydrozoa)

Large, free-floating crystals of calcium carbonate occur in vacuoles of gastrodermal cells of the hydroid Hydractinia symbiolongicarpus. Here, morphological details about the process by which these cells accumulate and sequester calcium are provided by a cytochemical method designed to demonstrate calcium at the ultrastructural level. Electron-dense material presumably indicative of the presence of calcium was EGTA-sensitive and was shown by parallel electron energy loss spectroscopy (EELS) and energy spectroscopic imaging (ESI) to contain calcium. Calcium occurred in only one cell type, the endodermally derived gastrodermal cell. In these cells, the electron-dense material appeared first as a fine precipitate in the cytosol and nucleus and later as larger deposits and aggregates in the vacuole. During the life cycle, gastrodermal cells of the uninduced planula and the planula during metamorphic induction sequestered calcium. In primary polyps and polyps from established colonies, gastrodermal cells sequestered calcium, but the endodermal secretory cells did not. Our observations support the hypothesis that gastrodermal cells function as a physiological sink for calcium that enters the organism in conjunction with calcium-requiring processes such as motility, secretion, and metamorphosis.     

The fine structure of the hypostome and mouth of hydra

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.     

Leucine aminopeptidase of the human blood flukes, Schistosoma mansoni and Schistosoma japonicum

An array of schistosome endoproteases involved in the digestion of host hemoglobin to absorbable peptides has been described, but the exoprotease responsible for catabolising these peptides to amino acids has yet to be identified. By searching the public databases we found that Schistosoma mansoni and Schistosoma japonicum express a gene encoding a member of the M17 family of leucine aminopeptidases (LAPs). A functional recombinant S. mansoni LAP produced in insect cells shared biochemical properties, including pH optimum for activity, substrate specificity and reliance on metal cations for activity, with the major aminopeptidase activity in soluble extracts of adult worms. The pH range in which the enzyme functions and the lack of a signal peptide indicate that the enzyme functions intracellularly. Immunolocalisation studies showed that the S. mansoni LAP is synthesised in the gastrodermal cells surrounding the gut lumen. Accordingly, we propose that peptides generated in the lumen of the schistosome gut are absorbed into the gastrodermal cells and are cleaved by LAP to free amino acids before being distributed to the internal tissues of the parasite. Since LAP was also localised to the surface tegument it may play an additional role in surface membrane re-modelling.     

Phagocytic activities of the gorgonian coral Swiftia exserta

The cellular response component of body defense in gorgonians and other cnidarians is thought to be carried out by cells with phagocytic capabilities. To test for the phagocytic character of cells, the introduction of foreign particles was employed and observed in both living cells and histological preparations of the gorgonian coral Swiftia exserta. Observations of untreated tissues revealed normal cells and tissue morphologies. A microscopic observation of living cells following the introduction of particles in a cut revealed that only a mixed population of colorless cells phagocytized the particles. Also particles or clumps of particles were seen on the surface of the colorless cells. Subsequent histological observations allowed identity of colorless cells to be inferred as granular amoebocytes, ectodermal cells, and gastrodermal cells. Cells stained for localization of peroxidase (indicative of phagocytic activity) demonstrated the presence of peroxidase-positive cells. Histological preparations revealed that major phagocytosis of particles was associated with tissue trauma. When particles were introduced by means of a cut or inserted thread, phagocytic activity was detected within 2 h. However, it was confined to the granular amoebocytes in the immediate site of trauma. After 24 h, extensive phagocytosis spread throughout a relatively large area surrounding the wound. At that later time, phagocytic cell types included granular amoebocytes, epidermal cells, sclerocytes, mesogleal cells, and gastrodermal cells of the solenia. Observations suggest that trauma induces phagocytosis in cells not normally phagocytic in S. exserta. No localization of phagocytic cells and no mitotic cells were observed at either 2 or 24 h after particle introduction.     

Cell Death and Acid Phosphatase Activity in the Regenerating Planarian Polycelis tenuis Iijima

A combination of microscopical, cytochemical, and biochemical techniques have been employed to study the changes occurring during the first seven days of blastema formation and regeneration after decapitation in adult Polycelis tenuis worms. Fine structural data reveal evidence of cell fragmentation, selective cell deletion, and phagocytosis at and below the wound surface. Initially, (0–12 h regeneration) cell debris is phagocytosed by intact parenchymal and gastrodermal cells near the cut surface which is later sealed (24 h) by a stretching of marginal epidermal cells. Wound sealing is followed by a migration of newly differentiated rhabdite cells into the epithelium. Morphological evidence of a selective cell autolysis precedes evidence of an accumuluation of lipid and glycogen reserves in the parenchymal and gastrodermal cells and the later (48 h regeneration time) aggregation of undifferentiated mitotically active neoblasts beneath the wound.
Biochemical data reveal an early period of high acid phosphatase (p-nitrophenyl phosphatase and sodium-β-glycerophosphatase) activity 3–12 h after injury, followed by a further intense period of activity at 44–48 h after decapitation. The coincident cytochemical data show an increased level of acid phosphatase activity associated with cell lysis and death in the wound and blastema zone and also with the digestion of phagocytosed cell debris.     

Ultrastructural evidence for the presence of nerve cells in the gastrodermis of Hydra

Summary Two types of nerve cells, namely, neurosensory and neurosecretory cells have been identified and described in the gastrodermis of Hydra pseudoligactis. The morphological criteria used for the identification of gastrodermal nerves are based on those presented previously for epidermal nerves. The third type of nerve cell in the epidermis, ganglionic cells, was not observed in these studies. The distribution, function and origin of gastrodermal nerve cells are discussed briefly.With the technical assistance of Linda M. Bookman.     

Cnidarian Primary Cell Culture as a Tool to Investigate the Effect of Thermal Stress at Cellular Level

In the context of global change, symbiotic cnidarians are largely affected by seawater temperature elevation leading to symbiosis breakdown. This process, also called bleaching, is triggered by the dysfunction of the symbiont photosystems causing an oxidative stress and cell death to both symbiont and host cells. In our study, we wanted to elucidate the intrinsic capacity of isolated animal cells to deal with thermal stress in the absence of symbiont. In that aim, we have characterized an animal primary cell culture form regenerating tentacles of the temperate sea anemone Anemonia viridis. We first compared the potential of whole tissue tentacle or separated epidermal or gastrodermal monolayers as tissue sources to settle animal cell cultures. Interestingly, only isolated cells extracted from whole tentacles allowed establishing a viable and proliferative primary cell culture throughout 31 days. The analysis of the expression of tissue-specific and pluripotency markers defined cultivated cells as differentiated cells with gastrodermal origin. The characterization of the animal primary cell culture allowed us to submit the obtained gastrodermal cells to hyperthermal stress (+?5 and +?8 °C) during 1 and 7 days. Though cell viability was not affected at both hyperthermal stress conditions, cell growth drastically decreased. In addition, only a +?8 °C hyperthermia induced a transient increase of antioxidant defences at 1 day but no ubiquitin or carbonylation protein damages. These results demonstrated an intrinsic resistance of cnidarian gastrodermal cells to hyperthermal stress and then confirmed the role of symbionts in the hyperthermia sensitivity leading to bleaching.     

Ratiometric imaging of gastrodermal lipid bodies in coral–dinoflagellate endosymbiosis

Cnidaria–dinoflagellate endosymbiosis is the phenomenon of autotrophic symbionts living inside the gastrodermal cells of their animal hosts. The molecular mechanism that regulates this association remains unclear. Using quantitative microscopy, we now provide evidence that the dynamic lipid changes in gastrodermal “lipid bodies” (LBs) reflect the symbiotic status of the host cell and its symbiont in the hermatypic coral Euphyllia glabrescens. By dual-emission ratiometric imaging with a solvatochromic fluorescent probe, Nile red (9-diethylamino-5H-benzo[α]phenoxazine-5-one), we showed that the in situ distribution of polar versus neutral lipids in LBs in living gastrodermal cells and symbionts can be analyzed. The ratio of Nile red fluorescence at red (R) versus green (G) wavelength region (i.e., R/G ratio) correlated with the relative molar ratio of polar (P) versus neutral (NP) lipids (i.e., P/NP ratio). The R/G ratio in host LBs increased after bleaching, indicating a decrease in neutral lipid accumulation in gastrodermal cells. On the other hand, neutral lipid accumulation inside the symbiont LBs resulted in gradual decreases of the R/G ratio as a result of bleaching. In comparison with the bleaching event, there was no relative lipid concentration change in host LBs under continual light or dark treatments as shown by insignificant R/G ratio shift. Patterns of R/G ratio shift in symbiont LBs were also different between corals undergoing bleaching and continual light/dark treatment. In the latter, there was little lipid accumulation in symbionts, with no resulting R/G ratio decrease. These results, demonstrating that the symbiotic status positively correlated with morphological and compositional changes of lipid bodies, not only highlight the pivotal role of LBs, but also implicate an involvement of lipid trafficking in regulating the endosymbiosis.     

Scanning electron microscopy of the muscle system of Hydra magnipapillata

The fine structure of the ectodermal and endodermal muscle layers of Hydra magnipapillata has been analyzed by scanning electron microscopy after hydrolytic removal of the mesoglea with NaOH and subsequent exposure of the basal and lateral aspects of the layers by mechanical dissection. The ectodermal muscle layer consists of fibrous processes of epithelial cells extending longitudinally to the body axis, whereas the endodermal muscle layer comprises cells with hexagonal bases and several strands of myonemes oriented circularly. In each layer, the muscular elements tightly interdigitate, extending a continuous muscle sheet along the mesoglea. The ectodermal and endodermal muscle sheets communicate with each other via foliate microprojections penetrating the mesoglea. On the lateral aspect of the ectodermal epithelium, spiny nerve fibers run along the upper surface of the muscle processes. The spines are often attached to muscle processes, suggesting that the former monitor muscle contraction. Nerve fibers occasionally come into contact with the mesoglea through narrow gaps between the muscle processes. In the hypostomal ectoderm, a small spindle-shaped cell, probably sensory in nature, extends an apical cilium and a long basal process.     

Regeneration by dissociated adult Hydra cells: a histologic study

Adult Hydra attenuata with vitally stained gastrodermal cells were dissociated into their component cells which were then randomly reaggregated into pellets by low-speed centrifugation. Representative examples of these preparations, which develop into normal adult hydra if left undisturbed, were examined fresh at low magnification and at higher magnification in fixed, stained, and sectioned specimens. The actual pellet stage lasts less than 1 hour because the adult ectodermal and gastrodermal cells rapidly sort themselves into an inner and outer layer and seem to secrete a new mesoglea immediately thereafter. The "embryo" becomes trilaminar and attains a central cavity by extruding a large amount of cellular debris at the end of the first day. At about this same time, new tentacles begin to differentiate from rapidly dividing and undifferentiated interstitial cells. Regulation of tentacle number and position occurs at the end of two days, and the body form is essentially reestablished within 60 hours by further differentiation of the hypostomes and body wall. Complete separation of the preparation into individual polyps does not occur until about 190 hours of development.     

Ultrastructural features of the trophonema and oogenesis in the starlet sea anemone, Nematostella vectensis (Edwardsiidae)

Abstract. The starlet sea anemone, Nematostella vectensis Stephenson 1935, is a burrowing, estuarine species that has become a model organism for fundamental studies of cnidarian and metazoan development. During early oogenesis, oocytes appear in the basal region of the gastrodermis in the reproductive mesenteries and gradually bulge into the adjacent connective tissue space (mesoglea) where the majority of oocyte growth and vitellogenesis occurs. However, oocytes do not physically contact the cellular and amorphous matrix of the mesogleal compartment due to a thin, intervening basal lamina. Oocytes retain limited contact with the basal gastrodermal epithelium via groups of ultrastructurally modified gastrodermal cells called trophocytes. Trophocytes are monociliated accessory cells of somatic origin that collectively form a structure called the trophonema, a unique accessory cell/oocyte association not observed outside the Cnidaria. The trophonema consists of 50–60 trophocytes that maintain contact with <1% of the oocyte surface and forms a circular, bowel‐shaped depression on the luminal surface of the gastrodermis as they sink into the mesoglea with the oocyte. The oocyte remains highly polarized throughout oogenesis with the germinal vesicle positioned near the trophonema and presumably representing the future animal pole of the embryo. Contact between the trophonema and the oocyte is restricted to cell junctions connecting peripheral trophocytes and narrow extensions from the oocyte. Previous studies suggest that the trophonema plays a role in transport of extracellular digestive products from the gastrovascular cavity to the oocyte, and the ultrastructural features described in this study are consistent with that view. Vitellogenesis is described for the first time in a sea anemone. Yolk synthesis involves both autosynthetic and heterosynthetic processes including the biosynthetic activity of the Golgi complex and the uptake of extraoocytic yolk precursors via endocytosis, respectively.     

Separation of cell layers in hydra

Summary The mesolamella of hydra is dissociable by means of divalent metal chelation with EDTA and is more susceptible to this treatment than the intercellular cements of each layer. Uncontaminated gastrodermal and epidermal cells may be obtained by this treatment.     

Increased susceptibility of algal symbionts to photo-inhibition resulting from the perturbation of coral gastrodermal membrane trafficking

The stability of cnidarian-dinoflagellate endosymbioses is dependent upon communication between the host gastrodermal cell and the symbionts housed within it. Although the molecular mechanisms remain to be elucidated, existing evidence suggests that the establishment of these endosymbioses may involve the sorting of membrane proteins. The present study examined the role of host gastrodermal membranes in regulating symbiont (genus Symbiodinium) photosynthesis in the stony coral Euphyllia glabrescens. In comparison with the photosynthetic behavior of Symbiodinium in culture, the Symbiodinium populations within isolated symbiotic gastrodermal cells (SGCs) exhibited a significant degree of photo-inhibition, as determined by a decrease in the photochemical efficiency of photosystem II (F _v/F _m). This photo-inhibition coincided with increases in plasma membrane perturbation and oxidative activity in the SGCs. Membrane trafficking in SGCs was examined using the metabolism of a fluorescent lipid analog, N-[5-(5,7-dimethyl boron dipyrromethene difluoride)-1-pentanoyl]-D-erythro-Sphingosylphosphoryl-choline (BODIPY-Sphingomyelin or BODIPY-SM). Light irradiation altered both membrane distribution and trafficking of BODIPY-SM, resulting in metabolic changes. Cholesterol depletion of the SGC plasma membranes by methyl-??-cyclodextrin retarded BODIPY-SM degradation and further augmented Symbiodinium photo-inhibition. These results indicate that Symbiodinium photo-inhibition may be related to perturbation of the host gastrodermal membrane, providing evidence for the pivotal role of host membrane trafficking in the regulation of this environmentally important coral-dinoflagellate endosymbiosis.     

Ultrastructure and cytochemistry of glycogen-containing vacuoles in gastrodermal cells in developing hydranths of a hydromedusan coelenterate

Hydranth buds from the colonial hydroid Sertularia pumila (Hydromedusae) were observed by electron microscopy during their development. Before hydranth expansion, the gastrodermal columnar digestive cells had large numbers of vacuoles. These vacuoles contained many membranous components as well as α-glycogen and dense ring- and crescent-shaped bodies. The rings and crescents were not osmiophilic, but did react to periodic acid oxidation in the PA-TSC-SP test for carbohydrate. These structures were digestible by α-amylase and pullulanase. The chemical analyses and the close association of the rings and crescents to α-glycogen particles showed that they may be a highly condensed form of glycogen. Golgi bodies in association with the gastrodermal vacuoles had acid phosphatase activity. This enzyme was only slightly active in the vacuoles. It is suggested that the vacuoles arc primarily storage organelles with a potential for digestion.     

Ultrastructure of the tentacle nerve plexus and putative neural pathways in sea anemones

Abstract. Neurons of sea anemone tentacles receive stimuli via sensory cells and process and transmit information via a plexus of nerve fibers. The nerve plexus is best revealed by scanning electron microscopy of epidermal peels of the tentacles. The nerve plexus lies above the epidermal muscular layer where it appears as numerous parallel longitudinal and short interconnected nerve fibers in Calliactis parasitica . Bipolar and multipolar neurons are present and neurites form interneuronal and neuromuscular synaptic contacts. Transmission electron microscopy of cross sections of tentacles of small animals, both C. parasitica and Aiptasia pallida , reveals bundles of 50–100 nerve fibers lying above groups of longitudinal muscle fibers separated by intrusions of mesoglea. Smaller groups of 10–50 slender nerve fibers are oriented at right angles to the circular muscle formed by the bases of the digestive cells. The unmyelinated nerve fibers lack any glial wrapping, although some bundles of epidermal fibers are partially enveloped by cytoplasmic extensions of the muscle cells; small gastrodermal nerve bundles lie between digestive epithelial cells above their basal myonemes. A hypothetical model for sensory input and motor output in the epidermal and gastrodermal nerve plexuses of sea anemones is proposed.