Showing papers on "Ultrastructure published in 1998"

The Peripheral Olfactory Organ of the Zebrafish, Danio rerio: an Ultrastructural Study

Abstract: The peripheral olfactory organ of the adult zebrafish, Danio rerio, was investigated by light as well as scanning and transmission electron microscopy. The olfactory organ consists of several lamellae that insert into a midline raphe, thus forming an oval-shaped rosette. Sensory and nonsensory regions are located separately on each lamella. The olfactory epithelium contains three types of receptor cells: two as described classically for other fishes, bearing either cilia or microvilli, the third being a new sensory cell type--the crypt cell. The crypt cell has no olfactory knob but bears microvilli as well as submerged cilia. Its axon travels together with the axons of the other receptor cells towards the basal lamina. Whereas the classical receptor cells are separated by supporting cells with small protrusions on their apical surfaces, the crypt cell is always surrounded by one or two specialized electron-lucent supporting cells which also bear microvillous-like apices. The nonsensory areas contain the goblet cells, ciliated nonsensory cells and epidermal cells with microridges.

Ultrastructure of the Olfactory Organ in the Clawed Frog, Xenopus laevis, During Larval Development and Metamorphosis

Abstract: Development of the olfactory epithelia of the African clawed frog, Xenopus laevis, was studied by scanning and transmission electron microscopy. Stages examined ranged from hatching through the end of metamorphosis. The larval olfactory organ consists of two chambers, the principal cavity and the vomeronasal organ (VNO). A third sensory chamber, the middle cavity, arises during metamorphosis. In larvae, the principal cavity is exposed to water-borne odorants, but after metamorphosis it is exposed to airborne odorants. The middle cavity and the VNO are always exposed to waterborne odorants. Electron microscopy reveals that in larvae, principal cavity receptor cells are of two types, ciliated and microvillar. Principal cavity supporting cells are also of two types, ciliated and secretory (with small, electron-lucent granules). After metamorphosis, the principal cavity contains only ciliated receptor cells and secretory supporting cells, and the cilia on the receptor cells are longer than in larvae. Supporting cell secretory granules are now large and electron-dense. In contrast, the middle cavity epithelium contains the same cell types seen in the larval principal cavity. The VNO has microvillar receptor cells and ciliated supporting cells throughout life. The cellular process by which the principal cavity epithelium changes during metamorphosis is not entirely clear. Morphological evidence from this study suggests that both microvillar and ciliated receptor cells die, to be replaced by newly generated cells. In addition, ciliated supporting cells also appear to die, whereas there is evidence that secretory supporting cells transdifferentiate into the adult type. In summary, significant developmental additions and neural plasticity are involved in remodeling the olfactory epithelium in Xenopus at metamor

Sampling the light-organ microenvironment of Euprymna scolopes: description of a population of host cells in association with the bacterial symbiont Vibrio fischeri.

Abstract: The symbiosis between the squid Euprymna scolopes and the luminous bacterium Vibrio fischeri has a pronounced diel rhythm, one component of which is the venting of the contents of the light organ into the surrounding seawater each day at dawn. In this study, we explored the use of this behavior to sample the microenvironment of the light-organ crypts. Intact crypt contents, which emerge from the lateral pores of the organ as a thick paste-like exudate, were collected from anesthetized host animals that had been exposed to a light cue. Microscopy revealed that the expelled material is composed of a conspicuous population of host cells in association with the bacterial symbionts, all of which are embedded in a dense acellular matrix that strongly resembles the bacteria-based biofilms described in other systems. Assays of the viability of expelled crypt cells revealed no dead bacterial symbionts and a mixture of live and dead host cells. Analyses of the ultrastructure, biochemistry, and phagocytic activity of a subset of the host cell population suggested that some of these cells are macrophage-like molluscan hemocytes.

Ultrastructural analysis of the sporozoite of Cryptosporidium parvum

Abstract: Cryopreparation of live sporozoites and oocysts of the apicomplexan parasite Cryptosporidium parvum, followed by transmission electron microscopy, was undertaken to show the 3D arrangement of organelles, their number and distribution. Profiles of parasites obtained from energy-filtering transmission electron microscopy of serial sections provided 3D reconstructions from which morphometric data and stereo images were derived. The results suggest that sporozoites have a single rhoptry containing an organized lamellar body, no mitochondria or conventional Golgi apparatus, and one or two crystalline bodies. Micronemes were shown to be spherical, numerous and apically located, and to account for 0.8% of the total cell volume. Dense granules were less numerous, larger, accounted for 5.8% of the cell volume, and were located more posteriorly than micronemes. A structure juxtaposed to the nucleus with similarities to the plastid-like organelle reported for other members of the Apicomplexa was observed. The detailed analysis illustrates the advantages of cryopreparation in retaining ultrastructural fidelity of labile or difficult to preserve structures such as the sporozoite of Cryptosporidium.

Three-dimensional analysis of the senescence program in rice (Oryza sativa L.) coleoptiles: Investigations by fluorescence microscopy and electron microscopy

Abstract: The cytological sequence of senescence-related changes in coleoptiles of rice (Oryza sativa L. cv. Nippon-bare) was studied using fluorescence and electron microscopy. The coleoptiles reach full size 3 d after sowing, then rapidly senesce and wither completely by day 7. The interveinal region in cross-sections taken 1 mm from the tip of the coleoptile was selected for this analysis. Fluorescence microscopy using samples embedded in Technovit 7100 resin, electron microscopy and immunoelectron microscopy using DNA-specific antibodies were used to elucidate the sequence of senescence-related events. These occur in the following order: (i) degradation of the chloroplast DNA (cpDNA): (ii) condensation of the nucleus in conjunction with a decrease in the size of the dense-chromatin region, shrinkage of the chloroplast, degradation of ribulose-1, 5-bisphosphate carboxylase oxygenase, dilation of the thylakoid membranes, increase in size and number of osmiophilic globules, condensation of the cytoplasm; (iii) disorganization of the nucleus, degeneration of the tonoplast; (iv) complete loss of the cytoplasmic components, distortion of the cell wall, invasion of microorganisms into the intercellular spaces and ultimately into the cell itself. The mitochondria maintain their ultrastructural integrity and a constant level of mitochondrial DNA throughout senescence, In young mesophyll cells, invagination of the tonoplast into the vacuole frequently occurs. This occasionally includes cytoplasmic material, which is digested in the vacuole as senescence proceeds. Immunoelectron microscopy suggests that cpDNA degradation involves rough digestion first, rather than rapid, direct decomposition of the DNA into nucleotides. The fragmented cpDNA is then dispersed throughout the chloroplast and cytoplasm.

Ultrastructure of the oocysts, sporocysts, and sporozoites of Toxoplasma gondii

Abstract: Transmission and scanning electron microscopy were used to study the ultrastructure of the oocysts, sporocysts, and sporozoites of the VEG strain of Toxoplasma gondii and to compare the ultrastructure of sporozoites with tachyzoites (from the peritoneum of mice) and bradyzoites (from brain tissue cysts in mice). Oocysts were surrounded by a thin veil of finely reticulate material. The oocyst wall consisted of 3 layers and contained a previously unknown disc-shaped micropyle that appeared as a depression in the oocyst wall. The sporocyst contained 4 sporozoites and a residuum of lipid and amylopectin granules. The sporocyst wall was 3-layered with the innermost layer consisting of 4 curved plates held together at sutures by an interposed strip. Exposure to excysting fluid caused the interposed strip to separate from the curved plates, which curled inward releasing the sporozoites. Sporozoites had a posteriorly located nucleus and all the organelles typical for coccidian zoites. Sporozoites, tachyzoites, and bradyzoites had similar numbers of rhoptries but differed in the numbers and sizes of micronemes, dense granules, amylopectin granules, and lipid bodies.

Histology and ultrastructure of the gut of the tilapia (Tilapia spp.), a hybrid teleost.

Abstract: Summary The morphology of the intestine has been studied in a species of warm water fish, Tilapia spp., a hybrid teleost of notable economic importance. Light and electron microscope results show that the intestine is a relatively undifferentiated muscular tube lined with a simple columnar epithelium interspersed with goblet cells. The proximal region has a greater surface area, manifested by elongated mucosal ridges. The enterocytes are covered apically with uniform microvilli and exhibit the typical ultrastructural features of pinocytosis, namely extensive invaginations of the luminal plasma membrane and massive accumulation of vesicles in the apical cytoplasm. The distal intestine mucosa is thinner and less elaborately folded and consists of columnar cells with shorter and sparser microvilli. Their supranuclear cytoplasm contains abundant clear vacuoles. Numerous endocrine cells can also be seen. Regional cellular ultrastructural features are correlated with digestive functions.

Ultrastructural effects of date extract on Candida albicans

Abstract: The effect of Berhi date extract on the ultrastructure of Candida albicans was studied by scanning and transmission electron microscopy. Exposure of yeast to 5% (w/v) date extract showed evidence of weakening in the cell wall with indications of cell distortion and partial collapse in some cases as seen by scanning electron microscopy. Increasing the concentration of date extract (20%, w/v) led to more drastic damage to the yeast with cell lysis and concurrent leakage of cytoplasmic material with eventual cell death. Ultrastructural investigation showed irregular shapes of cells treated with date extract, with prominent effects on cell wall layers. Cell membranes lost their integrity, aggregation of the cytoplasmic contents and large detachment of plasmalemma from cell wall was observed in the treated cells. These results suggest that date extract may have multiple effects on Candida with an increasing potential of using it for prophylaxis purposes.

CHARACTERIZATION AND PHYLOGENETIC POSITION OF THE ENIGMATIC GOLDEN ALGA PHAEOTHAMNION CONFERVICOLA: ULTRASTRUCTURE, PIGMENT COMPOSITION AND PARTIAL SSU rDNA SEQUENCE

Abstract: The morphology, ultrastructure, photosynthetic pigments, and nuclear-encoded small subunit ribosomal DNA (SSU rDNA) were examined for Phaeothamnion confervicola Lagerheim strain SAG119.79. The morphology of the vegetative filaments, as viewed under light microscopy, was indistinguishable from the isotype. Light microscopy, including epifluorescence microscopy, also revealed the presence of one to three chloroplasts in both vegetative cells and zoospores. Vegetative filaments occasionally transformed to a palmelloid stage in old cultures. An eyespot was not visible in zoospores when examined with light microscopy, but small droplets, similar to eyespot droplets, were apparent beneath the shorter flagellum when cells were viewed with electron microscopy. Zoospores had two flagella that were laterally inserted in the cell approximately one-third of the cell length from the apex. The longer flagellum was directed anteriorly and the shorter flagellum was directed posteriorly. Electron microscopy revealed the presence of tubular tripartite flagellar hairs on the longer flagellum, but no lateral filaments were found on the tripartite hairs. The general organization of the flagellar root system was similar to that of zoospores belonging to the Xanthophyceae and Phaeophyceae. However, the transitional region of the flagella contained a transitional helix with four to six gyres. Microtubular root R1 consisted of six microtubules at its proximal end and one microtubule at its distal end. Roots R2 and R4 consisted of one microtubule each and root R3 consisted of two microtubules. No rhizoplast was found. Thin-layer chromatography revealed the presence of fucoxanthin, diadinoxanthin, neoxanthin, and heteroxanthin as well as chlorophylls a, c1 and c2. High-performance liquid chromatography revealed the presence of fucoxanthin, diadinoxanthin, diatoxanthin, heteroxanthin, and β,β-carotene as well as chlorophylls a and c. The complete sequence of the SSU rDNA could not be obtained, but a partial sequence (1201 bases) was determined. Parsimony and neighbor-joining distance analyses of SSU rDNA from Phaeothamnion and 36 other chromophyte algae (with two Oomycete fungi as the outgroup) indicated that Phaeothamnion was a weakly supported (bootstrap = <50%, 52%) sister taxon to the Xanthophyceae representatives and that this combined clade was in turn a weakly supported (bootstrap = <50%, 67%) sister to the Phaeophyceae. Based upon ultrastructural observations, pigment analysis, and SSU rDNA phylogenetic analysis, Phaeothamnion is not a member of the Chrysophyceae and should be classified as incertae sedis with affinities to the Xanthophyceae and Phaeophyceae.

Trichinella spiralis : induction of the basophilic transformation of muscle cells by synchronous newborn larvae

Abstract: The present ultrastructure observations apply to the same experimental groups that were investigated in the first part of the study. The results of electron microscopy investigations correspond to those obtained using light microscopy methods. The ultrastructure data demonstrated that 1-h-old sNBL (group I) penetrated into the sarcoplasm of the muscle cells and transformed it basophilically, finally creating the “nurse cell-muscle larva complex.” These larvae also caused transformation of the same muscle cells without being present in the sarcoplasm. The larvae of group II (9-h-old sNBL) preserved transformation potential as well, but it was less intensive. Not all NBL settled in the muscle cells; some of them remained in the intercellular spaces. Group II larvae present in the muscle cells underwent early degeneration and necrosis more often than did group I larvae; the inflammatory cell reactions in the vicinity of the larvae were more intensive. The basophilic transformation of muscle cells that did not contain larvae in their sarcoplasm was not intensive and often stopped at the level of cell nuclei. The larvae of group III (6-day-old sNBL) neither settled in the muscle cells nor transformed the cell sarcoplasm.

Light and Electron Microscopic Study of Changes in the Organization of the Cortical Actin Cytoskeleton During Chromaffin Cell Secretion

Abstract: Chromaffin cells cultured for 2 days were incubated in the absence or presence of 10 microM nicotine for 40 sec. Resting and stimulated cells were fixed and either prepared for fluorescence microscopy or treated with Triton X-100 to obtain cytoskeletons for ultrastructural studies. Electron microscopy of cytoskeletons revealed the presence of polygonal areas devoid of actin filaments only in nicotinic receptor-stimulated cells. Staining of these cytoskeleton preparations with rhodamine-phalloidin, a probe for filamentous actin, produced fluorescent patterns and three-dimensional images similar to those obtained from resting or stimulated intact cells prepared directly for fluorescence microscopy. Moreover, the percentage of stimulated cells showing disrupted cytoskeleton at the electron microscopic level was similar to the percentage of stimulated cells showing patched rhodamine fluorescence at the fluorescence microscopic level. In addition, cells stimulated with nicotine for 40 sec showed a fivefold increase in amine output and a significant decrease in F-actin levels. These results provide the first ultrastructural evidence for nicotinic receptor-evoked chromaffin cell F-actin disassembly and show that the rhodamine-phalloidin-unstained areas observed in fluorescence microscopy represent the areas devoid of filamentous actin observed at the electron microscopic level.

Ultrastructural studies of spores and hypha of a glomus species

Abstract: The ultrastructure of spores and hyphae of a Glomus isolate (INVAM-906/312); structure and development are described with both light and transmission electron microscopy. The spore wall is formed by two layers in one group; the outer is evanescent and formed by an amorphous matrix, whereas the inner wall layer is laminated and has fibrils that seem to be arranged in an arcuate pattern as observed in other glomalean spores. The spore has a more complex wall structure than hyphae. The content of both spore and hyphae also differs regarding the presence and distribution of lipid globules, vesicles, and other structures. Intracytoplasmic bacterium-like organisms (BLOs) were observed in spores and germ tubes; some of these BLOs were apparently undergoing binary fission.

Observation of ultrastructural changes in cultured retinal pigment epithelium following exposure to blue light

Abstract: · Background: The retina can be damaged by light even when levels of energy are well below the threshold for thermal damage, and the experimental damage of the retinal pigment epithelium (RPE) may be induced more easily by blue light than by longer wavelengths of visible light. The present study demonstrates the ultrastructural damage produced by exposure to blue light in cultured RPE. · Methods: Long-Evans rats were enucleated 8–10 days after birth for primary culture. One week after seeding, the monolayer culture of RPE cells was exposed to a cool blue light (wavelength = 440±10 nm) for 36 h (12 h/day, 3 days) at 2.0 mW/cm2. Transmission electron microscopy was used to compare the exposed RPE with the control. The entire experiment was repeated 3 times independently. · Results: The cytoplasm of the exposed RPE exhibited degenerative changes, such as large whorls of membrane, lamellar whorls and whorled inclusions. · Conclusion: The RPE cells can be damaged directly by blue light after excluding the possible influence of phagosomes. This primary culture of RPE can also serve as an in vitro model for the study of light damage to the RPE.

Morphological study of Trichophyton rubrum: ultrastructural findings after treatment with 4-amino-3-methyl-1-phenylpyrazolo-(3,4-c)isothiazole.

Abstract: The antifungal activity of 4-amino-3-methyl-1-phenylpyrazolo-(3,4-c)isothiazole was studied on Trichophyton rubrum. The compound, at concentrations between 20 and 10 microg ml(-1), induces a remarkable reduction in the growth and causes deep morphogenetic anomalies. The ultrastructural modifications have demonstrated that the compound targets the cell membrane of the fungus, breaking down not only the endomembrane system, but also the 'outer' membrane, with consequent extrusion of materials in the medium. The results suggests a mechanism of action similar to other azoles clinically utilized.

Ultrastructural and lanthanum tracer examination of rapidly resorbing rat alveolar bone suggests that osteoclasts internalize dying bone cells.

Abstract: Glutaraldehyde-formaldehyde fixed undecalcified alveolar bone from 7-day-old rats was prepared for light and electron microscopy. Colloidal lanthanum was used as an ultrastructural tracer, and both random and semi-serial sections were examined. Lanthanum penetrated the infoldings of the ruffled border and some nearby vacuoles and vesicles. The majority of vacuoles and vesicles were lanthanum-free. Some osteoclast profiles contained a large vacuole with a cell enclosed in its interior. The enclosed cell exhibited an irregular nucleus containing condensed peripheral chromatin, intact cytoplasmic organelles, conspicuous rough endoplasmic reticulum and large blebs on the cell surface. These features are characteristic of osteoblasts or bone-lining cells or immature osteocytes which may be undergoing apoptosis or necrosis. The observation of remnants of cellular structures within internalized osteoclast vacuoles, together with the above results, suggests that osteoclasts engulf and probably degrade dying osteoblasts/bone-lining cells or immature osteocytes.

[Molecular effects of a microbicidal substance on relevant microorganisms: electron microscopic and biochemical studies on povidone-iodine].

Abstract: The microbicidal activity of the broad spectrum antimicrobial agent povidone-iodine is due to the strong oxidizing effects of free iodine on functional groups of amino acids, nucleotides and double bonds of unsaturated fatty acids. While the chemical mechanism of action of PVP-iodine is well understood, the actual sequence of events on the cellular and molecular level that causes rapid cell death has not been fully understood. The aim of this study was to elucidate effects of povidone-iodine on cell ultrastructure by electron microscopy and to monitor changes in enzyme activity and nucleotide efflux. Staphylococcus aureus, E. coli and C. albicans, medically relevant gram-positive, gram-negative and yeast micro-organisms, served as models. In the presence of povidone-iodine, rapid partitioning of the cytoplasm and pronounced coagulation of nuclear material was noted. Especially C. albicans exhibited a rapid, dose-dependent "loosening" of the cell wall; cells remained intact without lysis, rupture or wall breakage. Changes in beta-galactosidase and nucleotide concentrations were measured in E. coli. A rapid and dose-dependent loss of cellular beta-galactosidase activity was found, with no increase in the supernatant; loss of cellular nucleotides corresponded with an increase in the supernatant. Electron microscopy and biochemical observations support the conclusion that povidone-iodine interacts with cell walls of micro-organisms causing pore formation or generating solid-liquid interfaces at the lipid membrane level which lead to loss of cytosol material, in addition to enzyme denaturation. The chemical mechanism of action explains the fact that povidone-iodine does never generate resistance in micro-organisms.

Study of the action of alpha-hederin on the ultrastructure of Candida albicans.

Abstract: alpha-Hederin, a saponin isolated from Hedera helix (L.) (Araliaceae), was tested on Candida albicans ultrastructure. The concentrations used were 6.25, 12.5, and 25 micrograms ml-1 for an exposure time of 24 h. Transmission electron microscopy observations indicated that compared with untreated control yeasts, alpha-hederin induced modifications of cellular contents and alterations of cell envelope with degradation and death of the yeasts. The impact of alpha-hederin on the biomembranes and in particular on the plasmalemma is discussed. The antifungal activity of alpha-hederin was confirmed, as was the minimal inhibitory concentration (25 micrograms ml-1).

Further evidence of species variation in mechanisms of epithelial cell loss in mammalian small intestine: ultrastructural studies on the reindeer (Rangifer tarandus) and seal (Phoca groenlandica).

Abstract: Ultrastructural studies were conducted on mechanisms of epithelial cell loss in the small intestine of seal and reindeer. Mechanisms maintaining epithelial integrity were distinguished from those that did not and the non-epithelial cell types involved were identified. Three types of cell extrusion were noted. In two, tight junctional integrity was preserved and anucleate apical cell fragments (rather than complete cells) were lost into the lumen. In reindeer (type 1), this involved creating large intercellular spaces extending from the preserved apical cap to the lamina propria and containing enterocyte debris probably phagocytosed by subepithelial macrophages. A variant of this process (type 2) involved the gradual shrinkage of individual cells, which became more electron-dense, and the in situ degeneration of their nucleated subapical portions. Degenerated cell fragments and membrane whorls were confined to narrow intercellular spaces between approximating adjacent healthy enterocytes. The mechanism of removal of these fragments was unclear. In both cases, the proximity of intraepithelial lymphocytes suggested that they were involved in cell targetting and killing. Evidence of apoptotic nuclei was not found but nucleated cell fragments could have been washed out of the lumen during tissue preparation. Type 2 cell loss was seen in both species, as was another mechanism (type 3) reminiscent of necrosis. In contrast to other mechanisms, this was accompanied by breaks in epithelial continuity following gradual loss of cell electron density and total or subtotal degradation of organelles and membranes. In seal, this terminated in the loss of an abnormal cell apex and exposure of the contents of the cell remnant to the lumen. In reindeer, all the cell remnants may have been extruded before total membrane degeneration but, in both species, the otherwise tight epithelial barrier was clearly breached. Again, intraepithelial lymphocytes were associated with sites of necrosis. These findings provide evidence for further species differences in mechanisms of epithelial cell extrusion and suggest that necrotic cell loss may be more common than previously admitted.

Ultrastructure of fish cells involved in cellular defences against Saprolegnia infections: evidence of non-leucocytic nature.

Abstract: Fish cells attached to hyphae were observed in brown trout Salmo trutta L. infected with Saprolegnia parasitica. Earlier studies with light microscopy indicated that these cells were lymphocytes and neutrophils and that they were involved in the defence mechanisms against Saprolegnia infections. However, using electron microscopy, we found these attached cells did not show leucocytic characteristics, but instead shared some ultrastructural features with filament-containing cells. The pressure of a dense mass of cytoplasmic filaments precludes the leucocytic nature of these cells. How these cells could be involved in cellular defences against fungi is discussed.