Showing papers on "Ultrastructure published in 1994"

A novel microtubule-based motor protein (KIF4) for organelle transports, whose expression is regulated developmentally.

Abstract: To understand the mechanisms of transport for organelles in the axon, we isolated and sequenced the cDNA encoding KIF4 from murine brain, and characterized the molecule biochemically and immunocytochemically. Complete amino acid sequence analysis of KIF4 and ultrastructural studies of KIF4 molecules expressed in Sf9 cells revealed that the protein contains 1,231 amino acid residues (M(r) 139,550) and that the molecule (116-nm rod with globular heads and tail) consists of three domains: an NH2-terminal globular motor domain, a central alpha-helical stalk domain and a COOH-terminal tail domain. KIF4 protein has the property of nucleotide-dependent binding to microtubules, microtubule-activated ATPase activity, and microtubule plus-end-directed motility. Northern blot analysis and in situ hybridization demonstrated that KIF4 is strongly expressed in juvenile tissues including differentiated young neurons, while its expression is decreased considerably in adult mice except in spleen. Immunocytochemical studies revealed that KIF4 colocalized with membranous organelles both in growth cones of differentiated neurons and in the cytoplasm of cultured fibroblasts. During mitotic phase of cell cycle, KIF4 appears to colocalize with membranous organelles in the mitotic spindle. Hence we conclude that KIF4 is a novel microtubule-associated anterograde motor protein for membranous organelles, the expression of which is regulated developmentally.

Life-cycle and functional cytology of the hepatopancreatic cells of Astacus astacus (Crustacea, Decapoda)

Abstract: The hepatopancreas of the freshwater crayfish Astacus astacus was reinvestigated by means of light and electron microscopy using refined techniques of tissue preservation. The results contribute significantly to the solution of controversial problems of the decapod hepatopancreas such as cell genealogy, cellular interdependences, elimination of senescent cells and functional interpretation of the cell types. The three mature cell types of the organ, R-, F- and B-cells, are shown to originate independently from embryonic E-cells which are located at the blind-ending tips of the hepatopancreatic tubules. The less abundant M-cells are supposedly of non-hepatopancreatic origin since they are also found in other epithelia of the digestive tract. Differentiating cells can be assigned at an early stage to one of the three hepatopancreatic cell lines if the ultrastructural appearance and distribution pattern of their organelles are used as distinguishing features. The most sensitive markers are the Golgi bodies which have a cell-specific architecture and secretion product not only in mature cells but also in early differentiating stages. Later conversion of one cell type into another, as has often been proposed in literature, does not occur. Senescent cells are preferably expelled from the epithelium at the junction of neighbouring hepatopancreatic tubules and at the antechamber which links the hepatopancreas to the main digestive tract. Cellular discharge in the antechamber occurs by sliding of the oldest parts of the hepatopancreatic epithelium across a particular antechamber epithelium that was thus far unknown. New ultrastructural findings are described with respect to the absorptive apparatus of nutrient absorbing R-cells, the formation of Golgi vesicles and retrieval of membranes in digestive enzyme synthesizing F-cells, and the involvement of Golgi body and endoplasmic reticulum in the formation of heterophagic vacuoles in B-cells. The discovery of these ultrastructural features enables a more sophisticated functional interpretation of the hepatopancreatic cells of Decapoda.

Interaction of Pseudomonas aeruginosa with human respiratory mucosa in vitro

Abstract: Pseudomonas aeruginosa commonly infects the airways of patients with cystic fibrosis and bronchiectasis. It produces several toxins that slow ciliary beat, stimulate mucus production and damage epithelium. It adheres to epithelial cells, damaged mucosa (in animal models), and mucus. However, little is known of the interaction of P. aeruginosa with intact human respiratory mucosa. We have studied the interactions of a nonmucoid clinical isolate of P. aeruginosa with adenoid tissue in a novel organ culture model with an air-mucosal interphase P. aeruginosa (5.9 +/- 0.9 x 10(6) colony-forming units (cfu)) was pipetted onto the organ culture surface, and incubated for 15 min, 1, 2, 4, 8, 12, 16, and 24 h, at 37 degrees C in 5% CO2 in a humidified atmosphere. Assessment has been made by transmission and scanning electron microscopy. Transmission electron microscopy (TEM) showed that uninfected organ cultures had normal ultrastructure. TEM of infected organ cultures at 8 h showed significant epithelial damage: 43.9 +/- 10% of cells extruding from the epithelial surface, 17.7 +/- 3% of cells with loss of cilia, 32.9 +/- 10.2% of cells with mitochondrial damage, and 11.6 +/- 3% of cells with cytoplasmic blebbing. P. aeruginosa only infrequently adhered to normal epithelium, but adhered to areas of epithelial damage and to basement membrane. Scanning electron microscopy (SEM) of organ cultures up to 2 h found P. aeruginosa only infrequently associated with mucus. SEM at 4 h revealed P. aeruginosa predominantly associated with mucus and extruded damaged epithelial cells, but also occasionally associated with cilia, and very occasionally with unciliated cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Diagnostic transmission electron microscopy of tumors : with clinicopathological, immunohistochemical, and cytogenetic correlations

Abstract: Part 1 Ancillary procedures for evaluating human tumours: transmission electron microscopy other types of electron microscopy immunohistochemistry and immunoelectron microscopy tumour cytogenetics molecular pathology. Part 2 The nucleus: chromosomes and chromatin nuclear envelope - fibrous lamina nucleolus nuclear configurations nuclear inclusions. Part 3 The cytoplasm: organelles inclusions intracytoplasmic fibrils - cytoskeleton. Part 4 The cell surface: cell membrane specializations cell junctions. Part 5 Extracellular constituents: collagen elastic fibers amyloid - amyloidoma psamomma bodies basement membrane. Part 6 Ultrastructural features of specific human neoplasms: with clinicopathologic, immunohistochemical, and cytogenetic correlations (103 sections).

Ultrastructure of a new marine sand-dwelling dinoflagellate, Gymnodinium quadrilobatum sp. nov. (Dinophyceae) with special reference to its endosymbiotic alga

Abstract: A new sand-dwelling dinoflagellate, Gymnodium quadrilobatum sp. nov., is described from the south coast of Natal, South Africa. The organism contains 20–30 chloroplasts, which are peripherally arranged, and a conspicuous eyespot. The motile cell is directly transformed into a non-motile cell and cell division is restricted to the non-motile phase. The motile cell has a typical gymnodinioid organisation, while the non-motile cell possesses a thick cell wall and develops the characteristic cell shape, which is reminiscent of a four-leaved clover. The chloroplasts of G. quadrilobatum belong to an endosymbiotic alga the cytoplasm of which is separated from the host (dinoflagellate) cytoplasm by a single unit membrane. Other organelles contained in the endosymbiont cytoplasm are the nucleus, ribosomes and mitochondria. The endosymbiont is thought to be a chromophyte alga. The eyespot is bounded by a triple membrane and is situated in the host cytoplasm. On the basis of the similarities in the ultrastructure of...

Integumental ultrastructure and color patterns in the iridescent copepods of the family Sapphirinidae (Copepoda: Poecilostomatoida)

Abstract: The ultrastructure of the integument of the sapphirinid copepods was studied by scanning and transmission electron microscopy. Samples were collected between 1991 and 1993 by plankton-net tows from the subtropical and tropical waters of the North Pacific. In all the seven species examined of Sapphirina and Copilia, a structure with multilayered platelets was found in the epidermal cells of the dorsal integument of the male. Each platelet is a regular hexagonal prism. The platelets form a plate with honeycomb arrangement within each epidermal cell. Just ventral to the dorsal cuticle, 10 to 14 plates are located parallel to each other and to the cuticle. The mean diameter and thickness of the platelets measured between 1.0 and 1.8 μm and 61 and 83 nm, respectively, for the four species. The specific coloration of seven species was examined with reflected and transmitted light. The iridescent color may be explained by the theory of multiple thin-layer interference in some species which are considered to have an ideal laminar structure, but for the other species, mechanisms from non-ideal systems, including pigment-thin layer interaction, may also be involved.

Ultrastructure of Mycobacterium tuberculosis

Abstract: Mycobacterium tuberculosis was an early object of study by electron microscopy, consonant with its importance as a human pathogen. This chapter describes what is understood of the ultrastructure of M. tuberculosis and points out problems of interpretation and areas where further investigation, probably involving the development of new techniques, is needed. Most bacteria do not possess the elaborate system of internal compartments found in eukaryotic cells, and the information obtained about mycobacteria by electron microscopy is primarily information about the envelope layers, a discussion of which forms the major part of the chapter. It also describes the interaction between bacterium and host cells and some aspects of the ultrastructure of this interaction. Membranes of mycobacteria, including M. tuberculosis, appear in ultrathin sections as classic bilayers, with two electrondense layers separated by a transparent layer. If the hypothesis is that the walls of M. tuberculosis and other mycobacteria form permeability barriers somewhat analogous to the outer membranes of gram-negative bacteria, then the space between the outer leaflet of the membrane and the wall forms a compartment analogous to the periplasmic space of gram-negative bacteria. The commonest type of host cell is the macrophage; inside this cell, the bacteria occur within vacuoles, apparently the phagosomes formed as the bacteria are engulfed by the cells.

Variations in mitochondrial ultrastructure and dynamics observed by high resolution scanning electron microscopy (HRSEM)

Abstract: Rat adrenal cortex was processed for high resolution scanning electron microscopy (HRSEM) to confirm tubular cristae, reported by transmission electron microscopy to be present in cortex mitochondria. Mitochondria in several other tissue and cell types were also observed and their ultrastructure confirmed by using three-dimensional, stereo, high resolution scanning electron microscopy. The mitochondria in rat and human hepatocytes as well as human skin fibroblasts grown in culture contained tubular cristae approximately 30 nanometers in diameter. The fibroblast mitochondria proved to be long, up to 46 micrometers and branching, as compared to those in liver which were spherical in shape. Cold adapted brown fat cells were packed with mitochondria, these containing plate or shelf-like cristae. Branched, rat striated muscle mitochondria were observed to curve around contractile protein filament bundles. The muscle mitochondrial cristae were found to be both tubular and plate-like, within the same mitochondrion. The ratio of tubular cristae to plate-like cristae varied considerably between muscle mitochondria. In order to use ultrastructural changes in mitochondria for differential diagnosis, and because 3D reconstruction of mitochondria based on transmission electron microscopy serial sections is severely limited in resolution, it is imperative to first develop a correct understanding of tissue specific, normal mitochondrial ultrastructure based on three-dimensional, HRSEM methods.

Ultrastructure of the olfactory organ of the newt, Cynops pyrrhogaster

Abstract: The ultrastructure of the nasal sacs of the Japanese newt, Cynops pyrrhogaster, was studied by scanning and transmission electron microscopy. The paired nasal sacs of the newt are dorsoventrally flattened with a lateral nasal sinus off the main cavity of each sac. Throughout each sac is a series of ridges and grooves. In the main cavity, sensory epithelium with ciliated and microvillous receptor cells lines the grooves, and a thin, ciliated non-sensory epithelium lines the ridges. Secretory glands are present in the lamina propria. In the lateral nasal sinus, the ridges are lined with a thick, non-ciliated sensory epithelium that lacks glands. This region resembles and may function as a primitive vomeronasal organ.

Ultrastructure of nonspecific cytotoxic cells in teleosts. I. Effector-target cell binding in a marine and a freshwater species (seabream: Sparus aurata L., and carp: Cyprinus carpio L.).

Abstract: Background: Fish cytotoxic effectors form a cell population whose ultrastructure and properties of conjugation with target cells have not been completely established. We report the ultrastructure of the non-specific cytotoxic cells in a seawater teleost (Sparus aurata L.) and compare it to a freshwater species (Cyprinus carpio L.). Methods: Blood leucocytes were incubated with HeLa or B16 melanoma cells. Samples were processed for transmission electron microscopic study. Results: Conjugates consisting of leucocytes binding targets were regularly observed after 30 min, 1 hr, or 2 hr of incubation. In both species leucocytes binding to targets showed ultrastructural features of either monocyte-like or lymphocyte-like cells. Monocyte-like cells usually appeared flattened against the targets and seemed to enclose fragments of the target to form cytoplasmic vesicles and the content of their scarce cytoplasmic granules seemed to be delivered into these vesicles. In the seabream lymphocyte-like cells, dense cytoplasmic granules occurred only occasionally, and neither microvilli nor cell processes were present at the contact areas with the targets. In the carp, the contacts were more numerous and formed regularly interdigitating contact areas and the lymphocytes showed granules with characteristic dense and fibrillar contents. Conclusions: We conclude that seabream and carp have a leucocyte cell population with ultrastructural features of either monocytes or lymphocytes showing nonspecific cytotoxic ability. © 1994 Wiley-Liss, Inc.

Ultrastructure of Ependymoma

Abstract: Twenty-six ependymomas were studied by light and electron microscopy. By electron microscopy, the acellular zones around small vessels in light microscopic sections were seen to be composed of large numbers of closely packed, filament-rich, cytoplasmic processes ringing small vessels. Lumina were consistently present but many of them were too small to be seen by light microscopy. The lumina contained slender, curving microvilli and variable numbers of cilia. Their bordering cells were connected by unusually long tight junctions. Electron microscopy can be useful to establish or confirm a diagnosis of ependymoma when the light microscopic appearance is atypical, or when the tumor arises in an unusual location. The clinical data on the 26 cases has been reviewed; follow-up information was available on 23 patients.

An ultrastructural study of the dorsal lingual epithelium of the rat snake, Elaphe quadrivirgata.

Abstract: The histological characteristics and ultrastructure of the dorsal lingual epithelium of the rat snake, Elaphe quadrivirgata, were investigated by light microscopy and scanning and transmission electron microscopy. Most of the surface of the bifurcated part of the tongue was relatively smooth. Dome-shaped, hemispherical bulges were compactly arranged on the epithelial cell surface of the basal area of this region. Intercellular borders were clearly recognizable as striations. Microridges were densely distributed on the epithelial cell surface of the lingual body. Intercellular borders were thickened. A keratinized layer was clearly visible in the epithelium of the anterior bifurcated area, namely, at the apex of the tongue. Although keratohyalin granules were not found in any layer of the epithelium in this area, the cells of the surface layer were filled with keratin filaments. The dorsal lingual epithelium of the posterior area, namely, the lingual body, did not show any evidence of keratinization. Each cell on the surface side still had a large, oval nucleus and intact organelles, such as mitochondria, rough endoplasmic reticulum, ribosomes, tonofibrils, and tonofilaments. Cellular interdigitation was evident between adjacent cells and clear microridges or microvilli were observed on the cell membranes on the free-surface side of cells located in the surface layer. The phylogenetic relevance of these findings is discussed.

Secretion of phenolic bodies following fertilisation in Durvillaea potatorum (Durvillaeales, Phaeophyta)

Abstract: Cytological events immediately following plasmogamy in Durvillaea potatorum are described. Eggs contain several types of cytoplasmic vesicle differing in size and appearance. Histochemical tests and measurements are used to characterise and distinguish different types of vesicle containing phenolic compounds, lipids and polysaccharides. Within 2 min of plasmogamy, small phenolic vesicles located just below the egg membrane undergo mass synchronised exocytosis. The contents of these vesicles are discharged as phenolic bodies on the outside of the membrane. Secretion of phenolic bodies precedes secretion of the primary zygote wall by several minutes. Limited secretion of phenolics also occurs in unfertilised eggs. Peripheral phenolic vesicles are distinguishable from physodes, which also contain phenolic compounds but which are significantly larger and tend to be localised around the egg nucleus. The possible functional significance of the phenolic bodies is discussed. Coated pits and vesicles are common in...

Histochemical and Cytological Characterizations of Mucosal and Connective Tissue Mast Cells of Mongolian Gerbils (Meriones unguiculatus)

Abstract: Mast cells in jejunum, skin, and tongue of Mongolian gerbils (Meriones unguiculatus) were examined in terms of their histochemical, enzyme-histochemical, and ultrastructural properties. When glycosaminoglycans of mast cells in jejunum and tongue were characterized by measuring the critical electrolyte concentration, the salt concentration at which 50% of mast cells could be stained was > 1.0 M for those in the jejunum and also in the tongue, indicating that mast cells in both sites contained heparin. Enzyme-histochemical study revealed that mast cells in the jejunum of Mongolian gerbils were strongly positive for chymase and tryptase, whereas those in tongue and skin were essentially negative for both proteases. By electron microscopy, granular morphology and distribution of surface micro folds of mast cells in the jejunum were different from those in the ear skin. After in vivo stimulation with compound 48/80, however, mast cells in both sites showed remarkable degranulation. From these results, although Mongolian gerbils also have distinct mast cell subsets, their phenotypic properties are different from those of the previously known mast cell subsets of other animals.

Fertilization in Nicotiana tabacum: cytoskeletal modifications in the embryo sac during synergid degeneration. A hypothesis for short-distance transport of sperm cells prior to gamete fusion

Abstract: The cytoskeletal organization of the embryo sac of tobacco (Nicotiana tabacum L.) was examined at maturity and during synergid degeneration, pollen-tube delivery and gamete transfer using rapid-frozen, freeze-substituted and chemically fixed material in combination with immunofluorescence and immunogold electron microscopy. Before fertilization, the synergid is a highly polarized cells with dense longitudinally aligned arrays of microtubules adjacent to the filiform apparatus at the micropylar end of the cell associated with major organelles. The cytoskeleton of the central cell is less polarized, with dense cortical microtubules in the micropylar and chalazal regions and looser, longitudinally oriented cortical microtubules in the lateral region. In the synergid and central cell, F-actin is frequently found at the surface of the organelles and co-localizes with either single microtubules or microtubule bundles. Egg cell microtubules are frequently cortical, randomly oriented and more abundant at the chalazal end of the cell actin filaments are associated with microtubules and the cortex of the egg cell. At 48 h after pollination and before the pollen tube arrives, the onset of degeneration is evident in one of the two synergids: the electron density of cytoplasmic organelles and the ground cytoplasm increases and the nucleus becomes distorted. Although synergids otherwise remain intact, the vacuole collapses and organelles degenerate rapidly after pollen-tube entry. Abundant electron-dense material extends from the degenerated synergid into intercellular spaces at the chalazal end of the synergid and between the synergids, egg and central cell. Rhodamine-phalloidin and anti-actin immunogold labeling reveal that electron-dense aggregates in this region contain abundant actin forming two distinct bands termed "coronas". This actin is part of a mechanism in the egg apparatus which appears to precisely position and facilitate the access of male gametes to the egg and central cell for fusion.

Ultrastructural changes during in vitro encystment of Blastocystis hominis.

Abstract: The morphological changes occurring inBlastocystis hominis at different time points following in vitro encystment were studied by electron microscopy. The following stages of the parasite were sequentially seen: (a) the amoebic form, which was irregular in shape, with a majority of the organelles being concentrated at the condensed cytoplasmic region; (b) the pre-cystic form, which was rounded and had an electron-dense material forming a homogeneous wall around the central body; and (c) the cystic form, which had a very prominent, thick osmiophilic electron-dense wall, within which there were many inclusions and possibly reproductive granules. The amoebic form appeared to be an intermediate stage between the vacuolar form and the pre-cystic form, as this stage allowed the parasite to ingest bacteria to enhance encystment. The pre-cystic stage had previously been shown in experimental infection to be infective. The role of the cystic stage in producing infection is currently being investigated.

Ultrastructure of the endophyte and localization of nifH transcripts in root nodules of Coriaria nepalensis Wall. by in situ hybridization.

Abstract: SUMMARY Studies on the root nodules of Coriaria nepalensis Wall. using light and electron microscopy revealed that infected cortical cells are enlarged and form a compact kidney-shaped region with an acentric stele. Within a single cell, the actinomycetous endophyte has branched hyphae and elongate vesicles. Hyphae penetrate the host cell wall in different directions. Inside the host cell, the hyphae are located in the peripheral region of the cytoplasm near the host cell whilst the vesicles are positioned towards the centre of the cell around the central vacuole. No septa formation was observed in the microsymbiont vesicles. Host cell mitochondria were aggregated near the hyphal/vesicular junction of the endophyte. In situ hybridization studies demonstrated that the nifH transcripts are unequally distributed over the infected cortical cells. In infected cells, nifH mRNA was located in the region occupied by the elongate vesicles of the endophyte near the host cell central vacuole. The results indicate that the elongate vesicles of the nodule endophyte of C. nepalensis are functionally identical to the spherical vesicles of the nodule endophyte of Alnus.

Ultrastructure and protein uptake of the embryonic trophotaeniae of four species of goodeid fishes (Teleostei: Atheriniformes).

Abstract: Embryos of most species within the viviparous teleost family Goodeidae develop characteristics perianal processes that are considered to be derivatives of the embryonic hindgut. These processes, termed trophotaeniae, are covered with an epithelium that is continuous with the absorptive epithelium lining the hindgut. Gestation is intraovarian, and trophotaeniae mediate the uptake of maternally provided nutrients into the embryo from the ovarian fluid. Ultrastructural examination of the trophotaeniae of four goodeid species reveals substantial diversity in the organization of the epithelium within the family. The trophotaeniae of Alloophorus robustus, Zoogoneticus quitzeoensis, and Ilyodon furcidens have morphological features associated with the endocytosis of macromolecules and can be shown to endocytose the exogenous protein tracer horseradish peroxidase (HRP) rapidly. The trophotaenial epithelia of these species differ from one another with respect to other morphological features such as cell height, organization of the brush border, and the complexity of the intercellular spaces. The trophotaeniae of Goodea atripinnis lack an endocytotic apparatus and do not endocytose HRP. However, the overall organization of G. atripinnis trophotaenial cells suggests a function as a transporting epithelium. The cells have a dense brush border, numerous mitochondria, and many mitochondria that are enveloped by lamellar sheets of intracellular membrane. Post-fixation with osmium and potassium ferrocyanide reveals a marked difference in the complexity of the subepithelial connective tissue. Alloophorus robustus and Z. quitzeoensis exhibit an extremely electron-dense ground substance containing many acellular components. Goodea atripinnis exhibits an electron-lucid ground substance with few acellular components. © 1994 Wiley-Liss, Inc.

Light and electron microscopical study of two types of endocrines cell in the midgut of the adult worker honeybee (apis mellifera).

Abstract: The occurrence, development and ultrastructure of two types of gut endocrine cell have been studied in the midgut of adult honeybees. These cells, one of a basal granular type and one of a vesicular type, are evenly distributed throughout the posterior three-quarters of the midgut. Each crypt complex contains one of each cell type, both of which may be derived from the same stem cells as the enterocytes. They already contain their respective secretory product while still in the nidus. Both reach the midgut lumen by a narrow apex and are therefore of the open type. The granular cells release their secretory granules at the cell base in a typical endocrine way. In young vesicular cells the secretory vesicles are released at the cell base and in the intercellular spaces. Old cells are still filled with vesicles when they are shed in the midgut lumen. This seems to indicate that these cells have both an endocrine (or paracrine) and an exocrine function, the latter apparently by holocrinc release.

Ultrastructure of melanocytes in the dark cell area of human vestibular organs: Functional implications of gap junctions, isolated cilia, and annulate lamellae

Abstract: Background: It is known that melanocytes exist in almost all parts of the inner ear, such as the cochlear duct, stria vascularis, Reissner's membrane, modiolus, vestibular organs in the region surrounding the cristae and maculae, semicircular canals, and pars rugosa of the endolymphatic sac. But there have been few studies using human materials, because of the difficulty of obtaining materials. We attempted to investigate the detailed ultrastructure of melanocytes in the vestibular organs of human inner ear. Methods: Eight surgical specimens obtained from patients with vestibular schwannoma were studied by light microscopy and electron microscopy. Results: Melanocytes were found in the subepithelial layer of the dark cell area. Melanocytes had round or spindle-shaped nuclei and clear cytoplasm with brown pigment granules. Besides melanocytes, there were melanophages, fibroblasts, and small blood vessels. Through electron microscopy we found melanocytes with round-shaped melanosomes in various stages of pigmentation, well-developed Golgi apparatus and endoplasmic reticulum in the cytoplasm, and many cytoplasmic processes. Gap junctions were occasionally found between the cytoplasmic processes. And there were pinocytotic vesicles just under the limiting membrane of melanocytes, and intermediate filaments were abundant in the cytoplasm. Isolated cilia of melanocytes, annulate lamellae, and fusiform banded structures in the connective tissue area around melanocytes were found. Conclusions: Melanocytes in human vestibular organs actively synthesize melanosomes. Frequent findings of isoalted cilia and fusiform banded structures and the incidental existence of annulate lamellae may be an indicator of this metabolically activated state of melanocytes. Moreover, monitoring environmental changes by isolated cilia, melanocytes in the human inner ear could act not only as one cell but also as a group to achieve their physiological functions by means of information transmission through gap junctions. © 1994 Wiley-Liss, Inc.

Ultrastructural analysis of the intestinal content of earthworm Lumbricus rubellus Hoffm. (Annelida, Lumbricidae).

Abstract: Gut content sections of the earthworm Lumbricus rubellus, the surrounding soil and casts were studied using transmission electron microscopy. Bacteria, actinomycetes, dead fungi and empty mycelia were observed to occur throughout the gut content. In the foregut and midgut of worms, some bacteria were found to be partially or totally lysed and digested, although were protected by polysaccharides and clay particles. Not only inactive resting forms (spores) or cells protected by capsular polysaccharides and clay particles and/or by plant cellular remnants, but also separate metabolically active and dividing cells were recovered especially in hindgut. The formation of new bacterial microaggregates was noticed in the posterior intestine as well.

Ultrastructure and development of a thick basement membrane-like layer in the anchoring villi of macaque placentas

Abstract: Background: Anchoring villi and cytotrophoblastic cell columns are important structural components involved in placental morphogenesis. We have previously described the presence of an unusual basement membrane-like layer (BMLL) that separates these placental compartments. The purpose of the present study was to identify developmental changes in the ultrastructure of the BMLL and to assess its changes in extracellular matrix composition over the course of gestation. Methods: Conventional techniques were used to examine macaque placental tissue by transmission electron microscopy. Standard immunoperoxidase methods were used to identify type IV collagen, laminin and fibronectin in paraffin sections. Results: Until day 35 of gestation the BMLL was 70–100 nm thick and appeared similar to basement membranes seen in other regions of the villus, although it usually lacked a lamina lucida along the surface adjacent to the cytotrophoblast cells. Immunohistochemistry revealed the presence of laminin and type IV collagen in the BMLL. By 53 days of gestation the BMLL had hypertrophied at the junction of the anchoring villus and cell column, measuring 2,000–5,000 nm in thickness. The BMLL retained immunoreactivity for laminin and type IV collagen. Ultrastructural examination revealed the presence of a new component in the form of 10 nm microfibrils. By 89 days of gestation the BMLL was not reactive for laminin or type IV collagen but otherwise maintained the structural organization seen at 53 days. No additional changes were observed in the BMLL during late pregnancy. Conclusions: The BMLL is a distinct extracellular matrix region that separates the distal aspect of the anchoring villus from the proximal portion of the cell columns. Evidence indicates that adjacent cytotrophoblast plays a prominent role in the production of the BMLL. The BMLL may serve to organize this complex tissue by separating fetal mesenchyme from cytotrophoblast cells that are proliferating, differentiating, and migrating. Modifications to the composition of the BMLL may indicate changes in the role this matrix plays in the development of the placenta. © 1994 Wiley-Liss, Inc.

Symbiotic vesicle ultrastructure in high pressure-frozen, freeze-substituted actinorhizae

Abstract: Using tissue stained en bloc with chromic acid or tissue prepared by high pressure-freezing and freeze-substitution, it was possible to analyze quantitatively the ultrastructure of symbiotic vesicle envelopes (SVE) inAlnus serrulata, Ceanothus americanus, Elaeagnus umbellata, andMyrica cerifera. The lamina measured about 4.7 nm in thickness in thin section. Despite diverse symbiotic vesicle morphology, the SVE thickness was similar in all of these symbioses: 36–71 nm, which corresponded to 6–15 laminae based on counts of chromic acid-stained SVEs. This similarity in structure suggests that a similar environmental signal regulates envelope thickness in the different root nodules. Based on previous studies, this is likely to be pO2. Three types of envelope morphologies were distinguished: (1) theAlnus-type (as inAlnus andElaeagnus), which had localized thickenings around the vesicle and had thickest dimensions over the stalk; (2) theCeanothus-type. characterized as a relatively uniform envelope over both vesicle and attached hypha, and (3) theMyrica-type, which had no stalk region and a basal SVE thickness of about six laminae throughout except where localized thickening occurred. Localized thickening of the SVE resulted from extra numbers of laminae being deposited, generally over regions where septa contacted the edge of the vesicle. Freeze-substituted symbiotic vesicles had a variety of novel structures that are poorly preserved in chemically-fixed tissue. A paracrystalline body inAlnus symbiotic vesicles may be composed of particles that also exist free in the symbiotic vesicle cytoplasm. In addition, a previously unknown complex at the base of theAlnus-type symbiotic vesicle and within its stalk was evident in freeze-substituted tissues.

Ultrastructure of the holdfast of Phrixocephalus cincinnatus (Wilson), a blood-feeding parasitic copepod of flatfishes.

Abstract: Phrixocephalus cincinnatus is a blood-feeding copepod parasite inhabiting the choroid of the eye of the Pacific sanddab Citharichthys sordidus off the coast of southern California. The present study examined the fine structure of the midgut using both light and electron microscopy. The anterior midgut or stomach was lined with a tall columnar epithelium. The apical surfaces of the cells formed vesicles that pinched off and entered the gut lumen. Posteriorly, the midgut consisted of 2 morphologically distinct cell types; vacuolar and nonvacuolar. These cells probably were different developmental and functional stages of a single kind of columnar epithelial cell. The nonvacuolar cells possessed basally located nuclei, rough endoplasmic reticulum, and long microvilli indicative of an absorptive function. The vacuolar cells were characterized by short microvilli, endocytic vesicles, primary and secondary lysosomes, well-developed rough endoplasmic reticulum, and Golgi apparatus, suggesting that they were active in endocytosis as well as the synthesis of digestive and lysosomal enzymes. Distension of the apical surface of vacuolar cells with enzyme-containing vesicles resulted in the liberation of the distal part of the cell into the gut lumen. Subsequent lysis of the cell membrane released enzyme-containing vesicles, suggesting that digestive enzymes are discharged by apocrine secretion of the vacuolar cells. Both vacuolar and nonvacuolar cells absorb nutrients resulting from the extracellular digestion of host blood.

Electron microscopy of surface structures ofBlastocystis sp. from different hosts

Abstract: Samples ofBlastocystis sp. obtained from humans, monkeys, pigs and chickens were examined by scanning electron microscopy and transmission electron microscopy to compare surface structures. The surface coat ofBlastocystis sp. cells from each host species showed some morphological variations, but these were not sufficiently different to allow judgement to be made on speciation. The surface structure morphology appeared similar for samples ofBlastocystis sp. from the same host species. The surface coat of the cultured human isolate ofB. hominis was much thinner than that of cells from fresh human faecal material, and the cell surface appeared to be smoother and without the small projections seen in the fresh forms. Bacteria were frequently found in association with the surface coat ofBlastocystis sp. from all fresh faecal material. Possible functions of the surface coat, especially in relation to protection against osmotic shock, are discussed.