Cell surface saccharides of Trypanosoma lewisi. I. Polycation-induced cell agglutination and fine-structure cytochemistry

1975 ◽  
Vol 19 (3) ◽  
pp. 621-644
Author(s):  
D.M. Dwyer
Keyword(s):  
Fine Structure ◽  
Cell Surface ◽  
Alcian Blue ◽  
Chondroitin Sulphate ◽  
Surface Membrane ◽  
Ruthenium Red ◽  
Surface Coat ◽  
Cell Agglutination ◽  
Trypanosoma Lewisi ◽  
Cationic Compounds

Trypanosoma lewisi bloodstream and culture forms were agglutinated differentially with low concentrations of the cationic compounds: ruthenium red, ruthenium violet, Alcian blue chloride, 1-hexadecylpyridinium chloride, lanthanum chloride, and cationized ferritin. The bloodstream form trypanosomes gave the highest agglutination levels with each of the compounds tested. Ruthenium red was the most effective inducer of cell agglutination among the several cations used. Trypsin-treated bloodstream forms were agglutinated less in the presence of ruthenium red than untreated controls. Ruthenium red-induced cell agglutination also was lowered with chondroitin sulphate and dextran sulphate, but not with alpha-D-glucose, alpha-D-mannose or with several methyl glycosides. Treatment of the bloodstream trypanosomes with alpha-amylase, dextranase, or neuraminidase had little effect on agglutination levels obtained with ruthenium red. Fine-structure cytochemical staining with ruthenium red, ruthenium violet, and Alcian blue-lanthanum nitrate was used to ascertain the presence and distribution of presumptive carbohydrates in the trypanosome cell surface. The extracellular surface coat of the bloodstream forms stained densely with each of the polycationic dyes. Trypsin treatment removed the surface coat from bloodstream trypanosomes; however, the surface membranes of the organisms were stained densely with the several dyes. Similar surface-membrane staining was obtained with the cationic compounds and the culture forms, which lack a cell surface coat. Cationized ferrin was used at the fine-structure level to visualize the negative surface charge present in the cell surface coat and external membrane of the several trypanosome stages. Results obrained from the agglutination and cytochemistry experiments indicate that complex polysaccharides are present in the surface membranes and cell surface coat of T. lewisi bloodstream forms. Similar conclusions also pertain to the surface membranes of the T. lewisi culture from trypanosomes. The carbohydrates probably represent glycopeptide and glycoprotein structural components of the surface membrane of this organism.

Cell surface saccharides of Trypanosoma lewis i. II. Lectin-mediated agglutination and fine-structure cytochemical detection of lectin-binding sites

1976 ◽  
Vol 22 (1) ◽  
pp. 1-19
Author(s):  
D.M. Dwyer
Keyword(s):  
Fine Structure ◽  
Cell Surface ◽  
Lectin Binding ◽  
Glycoside Hydrolases ◽  
Surface Coat ◽  
Con A ◽  
Cell Agglutination ◽  
Flagellar Membrane ◽  
Surface Localization ◽  
Coupled Reactions

Bloodstream (BSF) and culture forms (CF) of Trypanosoma lewisi were specifically agglutinated with the plant lectins concanavalin A (Con A), soybean agglutinin (SBA), wheat germ agglutinin (WGA), and fucose-binding protein (FBP). Lectin-mediated cell agglutination was inhibited, and reversed in the presence of specific lectin-binding saccharides. Cells were agglutinated randomly with all lectins suggesting a uniform distribution in the trypanosome cell surface of the lectin-binding saccharide ligands. The BSF and CF were not agglutinated with phytohaemagglutinin-M, phytohaemagglutinin-P, or influenza virions. Living trypsinized BSF, which lacked a surface coat, gave agglutination results with the lectins identical to those obtained with living intact BSF. Glutaraldehyde- or formalin-fixed intact and trypsinized BSF gave results similar to those obtained with living cells and SBA, WGA, and FBP. However, intact, fixed BSF gave much lower agglutination levels with Con A than trypsinized-fixed, living intact, or living trypsinized BSF cells. Intact and trypsinized living and fixed CF gave identical agglutination results with each of the lectins. Living and fixed cells treated extensively with the glycoside hydrolases alpha-amylase, dextranase, and neuraminidase gave results with the lectins identical to those obtained with untreated cells. Con A bound at the cell surface was visualized with an iron-dextran (Fe-Dex) conjugate. Dense iron marker particles were distributed randomly in the intact BSF surface coat. The Con A-bound Fe-Dex marker was present on the pellicular and flagellar membrane outer lamina of trypsinized BSF and intact CF cells. Horseradish peroxidase (HRPO)-diaminobenzidine (DAB) coupled reactions also were used to visualize surface-bound Con A. Dense Con A-HRPO-DAB deposits were present uniformly in the BSF surface coat, and on the membranes of trypsinized BSF and intact CF trypanosomes. SBA and WGA were conjugated to HRPO and these used in DAB-coupled reactions at the ultrastructure level. Results obtained with the HRPO-conjugated lectins were similar in surface localization and distribution to those obtained with the Con A-HRPO-DAB preparations. Treatment of BSF and CF with the several glycoside hydrolases produced no apparent enhanced or reduced reactivity for the lectins in any of the fine-structure cytochemistry experiments. The cumulative results indicate that ligands similar or identical to alpha-D-mannose, N-acetylgalactosamine, and N-acetylglucosamine, and alpha-L-fucose are constituents in the extracellular surface coat matrix of T. lewisi BSF. Similar conclusions also pertain to the pellicular and flagellar membrane ligands of the BSF and CF cells. Moreover, results obtained with the glycoside hydrolases and influenza virions suggest that the T. lewisi cell surface ligands are not associated directly with repetitively bonded alpha-I,4- and alpha-I,6-D-glucans or sialic acid moieties.

Cell Surface Glycoconjugates of Euglena gracilis (Euglenozoa): Modifications under Potassium and Magnesium Deficiency

1997 ◽  
Vol 52 (1-2) ◽  
pp. 33-41
Author(s):  
Angelika Preisfeld ◽  
Gabriele Scholten-Beck ◽  
Hans Georg Ruppel
Keyword(s):  
Cell Surface ◽  
Protein Content ◽  
Alcian Blue ◽  
Euglena Gracilis ◽  
Magnesium Deficiency ◽  
Ruthenium Red ◽  
Complete Medium ◽  
Cell Surface Changes ◽  
Cell Surface Glycoconjugates ◽  
Surface Changes

Abstract Biochemical and ultrastructural examinations on the pellicle of autotrophically grown Eu­glena gracilis were carried out after three days under potassium and magnesium deficiency. Cell-surface changes were detected by lectin assay. Compared to cells grown in complete medium, deficient cells become larger in shape, accompanied by rising carbohydrate, chloro­phyll and protein content, bind more and other lectin molecules: an increase of mainly galactose and N-acetylgalactosamine receptors was observed. Investigations with the mucilage stains alcian blue and ruthenium red indicated that mucilaginous material is released under deficient conditions, whereas the control cells show a strong precipitate of these stains well inside the cells beneath the pellicle.

Fine Structure of the Cell Surface Coat of Trypanosoma musculi Bloodstream and Metacyclic Forms Using the Thiosemicarbazide-Silver Proteinate Method

1987 ◽  
Vol 73 (2) ◽  
pp. 415
Author(s):  
Michel Roger ◽  
Simon Garzon ◽  
Henri Strykowski ◽  
Pierre Viens
Keyword(s):  
Fine Structure ◽  
Cell Surface ◽  
Surface Coat

scholarly journals Surface coat on the epithelium of developing palatine shelves in the mouse as revealed by electron microscopy

Development ◽  
1974 ◽  
Vol 31 (3) ◽  
pp. 683-692
Author(s):  
R. M. Greene ◽  
D. M. Kochhar
Keyword(s):  
Electron Microscopy ◽  
Fine Structure ◽  
Ruthenium Red ◽  
Entire Length ◽  
Surface Epithelium ◽  
Positive Staining ◽  
Surface Coat ◽  
In Vitro Experiments ◽  
Acid Mucopolysaccharides

The fine structure of the surface epithelium of developing palatine shelves in the mouse was studied from days 11 through 14 of gestation. Ruthenium red, a cationic stain used as an ultrastructural indicator of acid mucopolysaccharides, was employed to detect the presence of any surface coat. Positive staining was first observed on day 12 of gestation and was seen to be present throughout the period of shelf elevation and fusion. It was seen over medial and lateral surfaces as well as the inferior tip of vertical shelves. The surface coat was found to be present along the entire length of the shelf, extending superiorly up the medial and lateral epithelial borders until it abruptly disappeared. Since this surface coat first appeared approximately 48 h prior to shelf elevation, it is suggested that its appearance may be associated with the ability of palatine shelves to undergo fusion as shown by previous in vitro experiments. The time of acquisition by the shelves of this ‘fusing potential’ is also in the range of 48 h before shelf elevation.

Rectilinear particle arrays in freeze-fracture replicas of the surface membrane of Paramecium tetraurelia

1986 ◽  
Vol 83 (1) ◽  
pp. 269-291
Author(s):  
R.R. Preston ◽  
T.M. Newman
Keyword(s):  
Plasma Membrane ◽  
Cell Surface ◽  
Surface Membrane ◽  
Freeze Fracture ◽  
Dorsal Surface ◽  
Surface Coat ◽  
Paramecium Tetraurelia ◽  
Intramembranous Particle ◽  
Logarithmic Growth Phase ◽  
Culture Cycle

Freeze-fracture replicas of the plasma membrane of unfixed, uncryoprotected Paramecium tetraurelia bear large rectilinear arrays of 11 nm particles arranged in 7–11 parallel rows. The arrays are of sufficient size to leave impressions in replicas of the underlying outer alveolar membrane, and are apparent as parallel ridges in replicas of the surface coat of deep-etched cells. By noting the location of arrays in replicas of identified portions of the cortex of P. tetraurelia, it has been possible to map the distribution of arrays over the cell surface. The arrays are found primarily over the anterior surfaces of the cell, covering an area that extends from the preoral suture over the left adoral field and a large portion of the anterior dorsal surface. Freeze-fracture analyses of cells taken from a number of different stages of a culture cycle suggest that the particle arrays are not replicated as an integral part of the cortex during cell division, but are assembled and oriented in the membrane as the cells mature. The appearance of small intramembranous particle complexes in the plasma membrane of cells in logarithmic growth phase supports this hypothesis, possibly representing an assembly stage in the formation of the larger particle arrays. The facts that the particle arrays are apparent in replicas of the surface coat of cells, are found primarily at the anterior of the cell body, and have a highly specific orientation with respect to the cell surface, strongly suggest that they function as chemoreceptors in P. tetraurelia.

scholarly journals THE SARCOPLASMIC RETICULUM AND ITS ASSOCIATION WITH THE T SYSTEM IN AN INSECT

1967 ◽  
Vol 32 (3) ◽  
pp. 535-545 ◽  
Author(s):  
Martin Hagopian ◽  
David Spiro
Keyword(s):  
Fine Structure ◽  
Sarcoplasmic Reticulum ◽  
Cell Surface ◽  
Osmium Tetroxide ◽  
Surface Membrane ◽  
Transverse Tubular System ◽  
Leucophaea Maderae ◽  
Tubular System ◽  
Femoral Muscle ◽  
T System

The fine structure of the sarcoplasmic reticulum and the transverse tubular system of the femoral muscle of the cockroach, Leucophaea maderae, was studied after prefixation in glutaraldehyde, postfixation in osmium tetroxide, and embedding in Epon. The sarcoplasmic reticulum in this muscle reveals features not previously reported. The sarcoplasmic reticulum is abundant, consisting mainly of a fenestrated envelope which surrounds each myofibril at all levels in the sarcomere. This sarcoplasmic reticulum envelope is continuous transversally as well as longitudinally along the myofibrils. Dyadic junctions are formed by a single T system element which contacts the unfenestrated sarcoplasmic reticulum of adjacent myofibrils in an alternating manner at the ends of the A band. At the dyads, regularly spaced thickenings of the sarcoplasmic reticulum membranes bordering the dyadic spaces are noted. These thickenings, however, do not contact the T tubule membrane. Typical dyadic contacts also are seen between the cell surface membrane and sarcoplasmic reticulum. Z line-like material is seen in contact with the membranes of the cell surface and longitudinal branches of the T systems.

Fine Structure of the Ventral Disk Apparatus and the Mechanism of Attachment in the Flagellate Giardia Muris

1973 ◽  
Vol 13 (1) ◽  
pp. 11-41 ◽  
Author(s):  
D. V. HOLBERTON
Keyword(s):  
Fine Structure ◽  
Surface Membrane ◽  
Ventral Surface ◽  
Surface Coat ◽  
Suction Force ◽  
Sinusoidal Waveform ◽  
The Face ◽  
Ventral Disk ◽  
Host Epithelium ◽  
Duodenal Epithelium

The topography of the Giardia trophozoite is dominated by the large domed sucking disk of the ventral surface. Attached to the host duodenal epithelium, the rim of this disk penetrates the enteric surface coat and interdigitates with microvilli of the epithelial cells, approaching to within 20 nm of the host surface membrane. Distortion of the host brush border within the disk suggests an applied suction force. A mechanical explanation of disk action is sought in a detailed description of the fine structure of components of the ventral surface - but is found to be untenable. The disk is supported by a platform of modified 25-nm microtubules, linked to the ventral membrane by side arms and bearing heavily cross-linked vertical dense ribbons. It is argued that such is the architecture of rigidity rather than relative movement. Around the disk a mobile cytoplasmic flange is supported by 2 lateral plates of periodic substructure. The flange has no clear mechanical role in attachment; a likely evolutionary origin from a component of the anterior axonemal axis is suggested. The cavity of the ventral disk leads posteriorly through a portal into the ventrocaudal groove: a shallow depression that houses the ventral flagella. Observation of isolated living trophozoites suggests that attachment depends on the continuing activity of the ventral flagella, which normally beat synchronously in a sinusoidal waveform. Electron micrographs confirm that this waveform is maintained in situ on the host epithelium. Of the 4 pairs of flagella, the ultrastructure of the ventral flagella is notable for additional components in the flagellar shaft, including an intraflagellar dense rod linked to 3 axonemal doublets by fine connectives. From a consideration of analogous macroscopic systems, a preliminary hydrodynamic analysis is advanced in which the suction force of attachment follows from the pattern of fluid flow induced by the beating ventral flagella. The significance of the conclusion that cytoplasmic microtubules (or structures derived from them) apparently maintain cell shape in the face of an applied external force is discussed.

Ruthenium red staining of the endolymphatic sac in the guinea pig

1988 ◽  
Vol 102 (9) ◽  
pp. 760-765 ◽  
Author(s):  
Masaya Takumida ◽  
Dan Bagger-Sjöbäck ◽  
Helge Rask-Andersen
Keyword(s):  
Epithelial Cells ◽  
Cell Surface ◽  
Guinea Pig ◽  
Tight Junctions ◽  
Apical Cell ◽  
Staining Technique ◽  
Ruthenium Red ◽  
Endolymphatic Sac ◽  
Surface Coat ◽  
Ruthenium Red Staining

AbstractThe ultrastructure of the guinea pig endolymphatic sac was studied, using the ruthenium red staining technique. The dye stained the apical cell surface coat and the homogeneous substance in the luminal space of the endolymphatic sac, when introduced from the luminal side of the epithelium. It is suggested that the surface coat and homogeneous substance may play an important part in the endolymph regulatory mechanism in the endolymphatic sac. When ruthenium red was introduced from the subepithelial side, the basolateral surface of the epithelial cells usually became brightly stained in the absence of staining of the apical cell surface, due to the presence of the tight junctions. In some instances, however, the dye penetrated beyond the level of the tight junctions. Pinocytotic vesicles and larger vacuoles in the epithelial cells were also sometimes stained, both apically and near the lateral cell surface. These findings suggest that endolymph efflux mechanisms in the endolymphatic sac may involve the combined actions of a paracellular and transepithelial flow as well as a transcellular, vacuolar bulk flow.

Cytochemistry of the very early stages of in situ coagulation in Calpodes ethlius (Lepidoptera)

1977 ◽  
Vol 55 (11) ◽  
pp. 1767-1772 ◽  
Author(s):  
M. Neuwirth ◽  
J. Lai-Fook
Keyword(s):  
Alcian Blue ◽  
Larval Instar ◽  
Ruthenium Red ◽  
Surface Coat ◽  
Early Stages ◽  
Latex Beads ◽  
Defence Reactions

Coagulation and phagocytosis are defence reactions which insects use in response to the introduction of foreign particles into their hemocoel. Coagulation occurs within 10 s of injection of latex beads and ink particles into a midfifth larval instar of Calpodes ethlius. A stringy, flocculent coagulum forms, trapping the particles and hemocytes. The hemocytes then go on to phagocytize the particles.The coagulum stains with alcian blue and lanthanum and dialyzed iron. The surface coat of granular hemocytes stains differently after coagulation has taken place with the same stains and with ruthenium red. There is no corresponding change in staining of the surface coat of plasmatocytes.

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