Gametogenic cycles and reproduction in the beadlet sea anemone Actinia equina (Cnidaria: Anthozoa)

Sexual Actinia equina have been maintained in the laboratory. Monthly biopsies show that these animals have annual gametogenic cycles. Regular samples from several natural populations suggest that animals in the wild show similar gametogenic periodicity. In the laboratory, males release sperm, but females brood young, at least for a few weeks. Members of these broods are released sporadically, usually after a water change. Males and non-sexual adults kept in isolation for over a year release a few juveniles during and at the end of that time. These young are likely to be asexually reproduced. There appears to be geographical variation of anemone size and in the frequency of sexual individuals in populations round the British Coast, so that modes of reproduction may vary.     

Tropomyosin isoforms present in the sea anemone,Anthopleura japonica (Anthozoa,Cnidaria)

Five isoforms of tropomyosin, designated as TMa, TMb, TMc, TMd, and TMe, were detected in the sea anemone, Anthopleura japonica. The apparent molecular weights of these isoforms were estimated to be approximately 30 kD to 37.5 kD, and their pI values were approximately 4.55 (TMa and TMb) and 4.65 (TMc, TMd, and TMe). Although sea anemone tropomyosin isoforms have the ability to bind to rabbit skeletal muscle actin, they preferably bind to actin at higher concentrations of Mg(2+) (10-20 mM) and slightly lower pH (6.2-7.2) than those used in conventional conditions. Antigenic properties of sea anemone tropomyosin seemed to be considerably specific to each isoform. Distribution of tropomyosin isoforms in the sea anemone body was somewhat portion-specific. TMa, TMb, and TMe were detected similarly in the extracts from tentacle, oral disc, column, mouth, and pedal disc. Although TMc and TMd were detected abundantly in the tentacle extract and moderately in the column and mouth extracts, these components were not contained in the pedal disc extract and detected only faintly in the oral disc extract.     

Ultrastructure of nematocyst discharge in catch tentacles of the sea anemone Haliplanella luciae (Cnidaria: Anthozoa)

The mature nematocyst lies just beneath the cnidodyte plasma membrane. A microtubule array surrounds the nematocyst capsule just beneath the capsule tip. We propose that the array helps to hold the capsule at the cnidocyte cell surface until discharge. The undischarged capsule tip is sealed by three apical flaps, joined together along complex radial seams. The seams are filled with subunits that appear to bind the flaps together. Upon discharge, the flaps separate along the radial seams to permit thread eversion. The everted thread is lined on both sides by subunits that are stained by antimonate, indicating that they bind calcium. We suggest that, together, the subunits hold the uneverted thread in its folded and coiled configuration. Thread eversion would follow subunit uncoupling. The capsule and thread interiors of partially discharged nematocysts are stained by antimonate. In contrast, the capsule and thread interiors of fully discharged nematocysts are not stained by antimonate. Thus, nematocyst calcium might be injected into the target tissue where it is presumed to act in conjunction with nematocyst venom to promote cell death.     

Scanning electron microscopy of exposed surfaces of the porcine placenta

The three-dimensional development of the separated parts of the porcine placenta from 9 Danish Landrace sows at gestational stages from 20 to 100 days was studied. After cautious separation of the allantochorion and the endometrium in a 1 mM buffered ethylenediaminetetraacetic acid solution, the separated parts were processed for scanning electron microscopy by routine methods. The macroscopic enlargement resulted from primary and secondary circular folds or plicae, which were permanent on the maternal side, whereas they were mainly non-permanent on the fetal side. The areolar placenta and interareolar placenta with formation of permanent microscopic folding on both sides were described. The observations of the separated parts yielded new information on the development of surface enlargement during gestation and revealed a hitherto unknown regular architecture of the endometrium by the formation of parallel primary ridges or rugae with secondary ridges or rugae at their sides subdividing the maternal troughs or fossae. This configuration on the maternal side explains the transformation of the regular chorionic ridges from the 63-day stage into bulbous protrusions at the 100-day stage. Based on these observations the precise terminology used above was proposed.     

Scanning electron microscopy of olfactory rosette in sea trout

Summary Scanning electron microscopy has been employed to study the central axis and laminae of the olfactory rosette in adult sea trout (Salmo trutta trutta L.) caught in the River Umeälven when they were homing from sea.—Both flat sides of the primary laminae are secondarily folded all over their surface. In one organ there are about 200 secondary laminae usually arranged in longitudinal, parallel ridges crossing the surface of the primary laminae. Initially they are covered with sensory epithelium, but as the folds grow they become covered with an increasing area of indifferent ciliar epithelium with bushes of cilia separated by microvilli cells and goblet cells. Parts of the central axis and primary laminae have a nonciliar indifferent epithelium. The sensory epithelium has irregularly arranged cilia. Like those of the indifferent epithelium they have uniform thickness and granulated surface. The function of laminae, secretion and cilia is discussed.The author wish to acknowledge the technical facilities and assistance in the use of the scanning electron microscope to Jeolco Stockholm office. This research was supported by grants 2389-10, 2389-11 and 2389-13 from the Swedish Natural Science Research Council.     

Scanning electron microscopy of dissociated pancreatic acinar cell surfaces

Summary The pancreatic acinar cell surfaces have been studied by SEM with a dissection technique and correlated with results obtained by TEM. The SEM results demonstrate characteristic arrangement of microplicae which in some areas are densely packed.In many areas, the microplicae are distributed in such a manner that they create zones with typical geometrical shapes and show a relatively smooth surface.These smooth areas may coincide, as indicated by correlated TEM results, with the limits of intimate contact between adjacent acinar cells which, in turn, represent part of the junctional complex. Another aspect revealed by these SEM preparations concerns the presence of groups of densely packed microplicae, arranged in regular rows and distributed along some grooves and/or infoldings of the cellular surface. On the basis of SEM and TEM information, it is likely that these structures correspond to intercellular (and possibly, in some cases, intracellular) canaliculi which topographically form a kind of extensive microlabyrinthine arrangement running along all the cell sides.One final point revealed by fractured samples concerns the finding of spherical zymogen droplets within the vesicles of the Golgi complex. Because in many scanning images these vesicles appear connected by small openings, it is suggested that they may represent a system of intercommunicating chambers (vacuoles) through which the zymogen droplets can be continuously accumulated and discharged into the acinar lumen.     

Different nematocytes have different synapses in the sea anemone Aiptasia pallida (Cnidaria,Anthozoa)

Sea anemones feed by discharging nematocysts into their prey, but the pathway for control of nematocyst discharge is unknown. The purpose of this study was to investigate the ultrastructural evidence of neuro-nematocyte synapses and to determine the types of synaptic vesicles present at different kinds of nematocyst-containing cells. The tip and middle of tentacles from small specimens of Aiptasia pallida were prepared for electron microscopy and serial micrographs were examined. We found clear vesicles in synapses on mastigophore-containing nematocytes and dense-cored vesicles in synapses on basitrich-containing nematocytes and on one cnidoblast with a developing nematocyst. In addition, we found reciprocal neuro-neuronal and sequential neuro-neuro-nematocyte synapses in which dense-cored vesicles were present. It was concluded that : (1) neuro-nematocyte synapses are present in sea anemones, (2) different kinds of synaptic vesicles are present at cells containing different types of nematocysts, (3) synapses are present on cnidoblasts before the developing nematocyst can be identified and these synapses may have a trophic influence on nematocyst differentiation, and (4) both reciprocal and sequential synapses are present at the nematocyte, suggesting a complex pathway for neural control of nematocyst discharge. J. Morphol. 238:53–62, 1998. © 1998 Wiley-Liss, Inc.     

An ultrastructural study of larval settlement in the sea anemone Urticina crassicornis (Cnidaria,Actiniaria)

Laboratory-reared larvae of the sea anemone Urticina (= Tealia) crassicornis have been examined by electron microscopy prior to and following settlement on algal substrata. At 18 days postfertilization, the free-swimming planula larva measures about 600 μm long. A stomodaeal invagination occurs at the narrow end of the larva and connects with a solid mass of endoderm in the core region. The endoderm possesses septa with well-developed myonemes and is situated subjacent to a thin sheet of mesoglea. The uniformly ciliated ectoderm that constitutes the outer layer of the larva contains: (1) spirocysts, (2) nematocysts, (3) mucus, (4) three types of membrane-bound granules, (5) a basiepithelial nerve plexus, and (6) a few nongranular cells that may represent sensory neurons. Within several minutes after the introduction of the algal substratum, the planula characteristically directs its broadened aboral end toward the alga and secretes a refractile sheet of material. As the aboral end attaches to the substratum, the larva becomes noticeably shorter along its oral-aboral axis, presumably owing to the contractions of myonemes that are located within the endodermal septa. All three types of granules and the ectodermal mucoid substances are exocytosed during settlement, but spirocysts and nematocysts characteristically remain undischarged. Ovoid, PAS⁺ granules are believed to be at least partly responsible for adhesion, since these granules are concentrated at the aboral end prior to settlement and are somewhat similar in ultrastructure to putative viscid granules produced by other species. Contrary to a previous report based on light microscopy, no discrete sensory organ is evident in serial sections of the aboral ectoderm. The ability of planulae to detect suitable substrata appears to depend instead on sparsely distributed sensory cells that occur throughout the larval ectoderm.     

Scanning electron microscopy of the integumental surfaces of Schistosoma haematobium.

The integumental surfaces of critical point dried S. haematobium were studied by scanning electron microscopy at 34 to 8,000 magnifications. There are marked differences between the surface structures of male and female as well as from one part of the same parasite to another. The surface of the male schistosome is moderately rough while that of the female is relatively smooth. SEM reveals certain basic features such as spines in the oral sucker and the acetabulum of both sexes which may facilitate rasping and/or attachment of the parasite for residence in the bloodstream of the definitive host. The lining of the gynecophoral canal is roughened by minute spines. The presence of a gynecophoral fold may enhance anchorage of the female in the grasp of the male. The significance of visualization of surface features by SEM as a means for differentiating species is not yet known.     

A jellyfish-eating sea anemone (Cnidaria,Actiniaria) from Palau: Entacmaea medusivora sp. nov.

Entacmaea medusivora sp. nov., a member of the family Actiniidae, inhabits Jellyfish Lake in Palau, western Caroline Islands, and eats the rhizostome jellyfish Mastigias papua. The anemone is azooxanthellate, despite its jellyfish prey containing symbiotic algae. Well fed anemones released ciliated planula-like larvae in the laboratory when maintained at more than 30 °C. It could not be determined whether the larvae, which usually settled around the parent within 24 h of release, were produced sexually or asexually.     

Modifications of mammalian cell surfaces induced by sugars: Scanning electron microscopy

Substitution of galactose, xylose, or mannose for glucose in the growth medium of L cells or the addition of equal concentrations of the alternate sugars to glucose-containing medium results in marked morphologic changes. The changes are revealed by conventional staining for light microscopy and by scanning electron microscopy.L cells grow indefinitely on combinations of equal concentrations of glucose and galactose, xylose, or mannose. There appear to be no significant differences in growth rate on glucose compared to the combinations of sugars cited. Cells subcultured from glucose to the combinations while undergoing rapid multiplication show marked morphologic changes by light and scanning electron microscopy within 36 hr.Of particular note are the loss of microvilli; flattening of the cells; assumption of polygonal shape; prominence of nuclei and nucleoli; and changes in the structure and distribution of filopodia. Virtually all cells in the population exhibit the changes noted.     

Scanning electron microscopy of trichomes of cheilanthes (sinopteridaceae)

Brittonia - The trichomes of two problematic complexes in the genusCheilanthes were studied. In one group, the trichomes ofC. tomentosa are long, multicellular, uniseriate, and flat whereas those...     

A new species of sea anemone (Cnidaria: Anthozoa: Actiniaria) from Manus Basin hydrothermal vents, South-western Pacific

During the BIOACCESS Japanese cruises (1996 & 1998), active hydrothermalism and associated vent fauna were studied on the South-eastern Rift of Manus Basin (South-western Pacific). In the PACMANUS vent field, a conspicuous vent fauna was sampled, including an actinostolid sea anemone (Actiniaria) belonging to an undescribed genus and species. Pacmanactis hashimotoi gen. et spec. nov. is here described, and represents the 9th sea anemone reported from hydrothermal vents.     

Scanning electron microscopy of the labia of Baylisascaris procyonis (Nematoda)

The labial organization of adult Baylisascaris procyonis was studied by scanning electron microscopy, and found to be similar for males and females. The apical part of each lip was smooth, and the basal part reticulated. The dorsal lip possessed two dorsolateral double papillae and two internal labial papillae; the two subventral lips each had one ventrolateral double and one externolateral papilla, two internal labial papillae, and an amphid. The small papilla of each double set was dome-shaped and smooth, whereas the large papilla was broad and had a prominent central pore. The externolateral papillae had raised, highly sculptured surfaces with numerous slits and creases present. Amphids resembled those of previously studied nematodes. The internal labial papillae consisted of pits. Denticles arose as a single row from the apical edge of the inner labial surface, were usually evenly spaced, and pyramidal or conoidal in shape. They were typically unicuspid, but bicuspid denticles were occasionally seen. Denticle shape and size varied between specimens and on each specimen. A pit was seen in the cuticle in the central region of the denticular row of all lips. Several of these findings represent new information concerning ascaridoid nematodes.     

An ultrastructural study of the establishement of the testicular cysts during spermatogenesis in the sea anemone Actinia fragacea (Cnidaria: Anthozoa)

Spermatogenesis in the sea anemone Actinia fragacea takes place in numerous testicular cysts located in the mesoglea of the gonads. Prospermatogonia arise among the bases of the gonadal epithelial cells bordering the mesoglea, and later migrate into the mesoglea to establish the cysts. The prospermatogonia arise singly, but soon most are found as small groups within the endoderm. They are small cells, 6–7 μm in diameter, and have relatively large nuclei with a single nucleolus. Their cytoplasm is dense, and contains dense bodies and nuage material as well as Golgi, mitochondria, and individual cisternae of endoplasmic reticulum. Each prospermatogonium bears a flagellum, originating in a groove or channel in the cytoplasm. A small proportion of prospermatogonia enter the mesoglea singly, but most migrate as elongate groups or “slugs” of cells. As they enter, the groups often become constricted into hour-glass shapes, and they become covered by the endodermal basal lamina. During the later stages of entry, the last part of the group to enter retains contact with the bases of the epithelial cells, which are dragged into the mesoglea behind the germ cells. This contact between germ cells and endoderm persists throughout spermatogenesis and prevents closure of the mesoglea behind the group. The endodermal cells involved begin specialization to form the trophonema. Once entry is complete, the groups enlarge rapidly to form the testicular cysts. A small number of germ cells appear to remain behind in the endoderm after most have entered the mesoglea, and the possible significance of these cells is discussed.     

Scanning electron microscopy (SEM) of the chick embryo primitive streak

The structure of the cells forming the primitive streak was examined by SEM in a series of embryos at Hamburger and Hamilton's stages 2--5. Specimens were prepared by stripping the endoderm from fresh embryos in New Culture and by fracturing whole fixed embryos along and at right angles to the primitive streak. At all stages of examination the SEM appearance of cells within the privitive streak was quite different from that of ectodermal, endodermal or mesodermal cells away from the streak. Streak cells were closely packed, lay with their long axes directed from ectoderm to endoderm and possessed many flat leaf-like processes. By contrast the ectoderm formed a columnar epithelium, the endoderm a flat epithelium and the mesoderm was a layer of loosely arrangedcells with long. thin processes. Within the streak SEM did not show any differences between cells that could identify them specifically as future endoderm or mesoderm cells. It was concluded that during gastrulation all the cells migrating through the primitive streak have the same appearance regardless of their eventual destination in the embryo. This structure may be attributable to the type of movement made by cells during invagination.     

Scanning electron microscopy of dairy equipment surfaces contaminated by two milk-borne micro-organisms

Ethanol dehydration followed by argon replacement induced drying (ARID) was found to be a suitable method for the preparation of glass, stainless steel and rubber surfaces which had been in contact with inoculated milk and which were to be examined using scanning electron microscopy (SEM). This technique was used to examine samples of all three materials which had been subjected to both single and repeated inoculation with whole milk containing a Pseudomonas sp. or a Micrococcus sp. and incubated for various periods. Some samples were also prepared for SEM using a cryofixation technique. The Pseudomonas sp. was found to proliferate on glass and stainless steel surfaces but not on rubber. Due to the clumping tendency of the Micrococcus sp. proliferation of this organism was more difficult to assess accurately. In general there was no difference in results obtained between single and repeated inoculation. Various factors which may have aided attachment of micro-organisms to surfaces were identified viz. , surface channels present in stainless steel, milk deposits and the production of extracellular material. The value of using both the cryofixation and chemical preparatory techniques for the identification of artifacts is discussed.