Immunolocalization of a Schistosoma mansoni facilitated diffusion glucose transporter to the basal, but not the apical, membranes of the surface syncytium

Parasitology ◽  
1995 ◽  
Vol 110 (4) ◽  
pp. 383-394 ◽  
Author(s):  
C. Zhong ◽  
P. J. Skelly ◽  
D. Leaffer ◽  
R. G. Cohn ◽  
J. P. Caulfield ◽  
...  
Keyword(s):  
Schistosoma Mansoni ◽  
Glucose Transporter ◽  
Light And Electron Microscopy ◽  
Linear Structure ◽  
Electron Microscopic Examination ◽  
Cdna Clones ◽  
Facilitated Diffusion ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Internal Domain

SUMMARYAdult parasites of Schistosoma mansoni reside within vertebrate mesenteric veins where they consume immense quantities of host glucose after transporting the sugar through their surface syncytium or tegument. Previously we obtained cDNA clones encoding two functional facilitated diffusion glucose transporter proteins expressed by S. mansoni adult worms (Skelly et al. 1994). Antibodies specific for one transporter (SGTP1) have been generated against an extrafacial and an internal domain of the protein and used to localize the protein by light and electron microscopy. By light microscopy both antibodies stain a linear structure approximately 1–5 μm from the surface of the tegument of adult male and female schistosomes. Electron microscopic examination of frozen thin sections show binding of the antibodies to membranes in the base of the tegument and not to the membranes covering the outer surface or their invaginations. Analysis of the gold distribution suggests that the extrafacial domain is disposed toward the interstitial space beneath the tegument and the internal domain faces the syncytial plasm. The localization suggests that SGTP1 may function to transport free glucose from within the tegument and into the interstitial fluids that bathe the internal organs of these parasites.

Toxic effects of inhaled DMMP on the kidneys of Fischer-344 rats

1987 ◽  
Vol 45 ◽  
pp. 880-881
Author(s):  
D.R. Mattie ◽  
C.J. Hixson
Keyword(s):  
Left Kidney ◽  
Inhalation Exposure ◽  
Electron Microscopic Examination ◽  
Exposure Period ◽  
Male Rats ◽  
Continuous Exposure ◽  
Fischer 344 Rats ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Fischer 344

Dimethylmethylphosphonate (DMMP) is a simple organophosphate used industrially as a flame retardant and to lower viscosity in polyester and epoxy resins. The military considered the use of DMMP as a nerve gas simulant. Since military use of DMMP involved exposure by inhalation, there was a need for a subchronic inhalation exposure to DMMP to fully investigate its toxic potential.Male Fischer-344 rats were exposed to 25 ppm or 250 ppm DMMP vapor on a continuous basis for 90 days. An equal number of control rats were sham-exposed. Following the 90-day continuous exposure period, 15 male rats were sacrificed from each group. Two rats from each group had the left kidney perfused for electron microscopic examination. The kidneys were perfused from a height of 150 cm water with 1% glutaraldehyde in Sorensen's 0.1M phosphate buffer pH 7.2. An additional kidney was taken from a rat in each group and fixed by immersion in 2.5% glutaraldehyde and 2% paraformaldehyde in 0.1M cacodylate buffer pH 7.4. A portion of the 9 kidneys collected for electron microscopy were processed into Epon 812. Thin sections, stained with uranyl acetate and lead citrate, were examined with a JEOL 100B Transmission Electron Microscope. Microvilli height was measured on photographs of the cells of proximal tubules. This data, along with morphologic features of the cells, allows the proximal convoluted tubules (PCT) to be identified as being S1, S2, or S3 segment PCT.

scholarly journals Application of lectin--gold complexes for electron microscopic localization of glycoconjugates on thin sections.

1983 ◽  
Vol 31 (8) ◽  
pp. 987-999 ◽  
Author(s):  
J Roth
Keyword(s):  
Endoplasmic Reticulum ◽  
Concanavalin A ◽  
Binding Sites ◽  
Light And Electron Microscopy ◽  
Gold Complex ◽  
Electron Microscopic ◽  
Gold Complexes ◽  
Thin Sections ◽  
Renal Tubular Cells ◽  
Lowicryl K4m

A method is described for the electron microscopic detection of lectin-binding sites in different cellular compartments and extracellular structures that uses thin sections from resin-embedded tissues. Various lectins (Ricinus communis lectin I and II, peanut lectin, Lotus tetragonolobus lectin, Ulex europeus lectin I, Lens culinaris lectin, Helix pomatia lectin, and soybean lectin) were bound to particles of colloidal gold and used for direct staining of thin sections or glycoprotein--gold complexes were prepared and applied in an indirect technique (concanavalin A and horseradish peroxidase--gold complex; wheat germ lectin and ovomucoid--gold complex). The details for preparation of such complexes from 14 nm gold particles are reported. The conditions of tissue processing that gave satisfactory staining results and good fine structure preservation were mild aldehyde fixation without osmification and low temperature embedding with the hydrophilic resin Lowicryl K4M. None of the so-called etching procedures was necessary prior to labeling of Lowicryl K4M thin sections. Examples of the use of this approach for detection of glycoconjugates in the rough endoplasmic reticulum, Golgi apparatus, and mucin of intestinal goblet cells as well as plasma membrane and various intracellular structures of absorptive intestinal and renal tubular cells are shown. A comparison is made with preembedding staining results on Concanavalin A-binding site localization in rat liver which shows that problems of penetration common in such a technique are circumvented by the postembedding approach described here. Concanavalin A-binding sites were not only consistently found in nuclear envelope, rough and smooth endoplasmic reticulum, plasma membranes, and collagen fibers, but also in mitochondria, glycogen, ribosomes, and nucleus. These data and those of a previous investigation (Roth J, Cytochem 31:547, 1983) prove the applicability of this cytochemical technique for postembedding localization of glycoconjugates by light and electron microscopy.

Molecular and functional characterization and tissue localization of 2 glucose transporter homologues (TGTP1 and TGTP2) from the tapeworm Taenia solium

Parasitology ◽  
1998 ◽  
Vol 117 (6) ◽  
pp. 579-588 ◽  
Author(s):  
D. RODRÍGUEZ-CONTRERAS ◽  
P. J. SKELLY ◽  
A. LANDA ◽  
C. B. SHOEMAKER ◽  
J. P. LACLETTE
Keyword(s):  
Glucose Transporter ◽  
Cytochalasin B ◽  
Functional Characterization ◽  
Taenia Solium ◽  
Cdna Clones ◽  
Facilitated Diffusion ◽  
Porcine Cysticercosis ◽  
Isolation And Characterization ◽  
Hexose Uptake

Tapeworms absorb and consume large quantities of glucose through their syncytial tegument, storing the excess as glycogen. Although some studies on the metabolism of glucose in several tapeworms are available, the proteins that mediate its uptake and distribution in their tissue have not been identified. We describe the isolation and characterization of cDNA clones encoding 2 facilitated diffusion glucose transporters (TGTP1 and TGTP2) from Taenia solium, the causal agent of human and porcine cysticercosis. Radio-isotope labelled hexose uptake mediated by TGTP1 expressed in Xenopus oocytes is inhibited by the natural stereoisomers d-glucose and d-mannose but not by l-glucose. Transport by TGTP1 is sensitive to classical inhibitors of facilitated diffusion such as phloretin and cytochalasin B, and insensitive to ouabain. TGTP2 did not function in Xenopus oocytes. Localization studies using specific anti-TGTP1 and anti-TGTP2 antibodies show that TGTP1 is abundant in a number of structures underlying the tegument in adult parasites and larvae, whereas TGTP2 appears to be localized only on the tegumentary surface of the larvae and is not detected in adults.

Cation Content and Transport Characteristics of the Sickle-Cell Erythrocyte and their Relationship with Structural Changes in the Membrane

1974 ◽  
Vol 46 (6) ◽  
pp. 679-692 ◽  
Author(s):  
J. Kurantsin-Mills ◽  
M. Kudo ◽  
S. Kojo Addae
Keyword(s):  
Sickle Cell ◽  
Sodium Transport ◽  
Structural Changes ◽  
Electron Microscopic Examination ◽  
Active Sodium ◽  
Electron Microscopic ◽  
Potassium Efflux ◽  
Thin Sections ◽  
Sodium Efflux ◽  
Active Sodium Transport

1. The intra-erythrocytic concentrations of sodium and potassium and the water content have been determined for haemoglobin (Hb) SS cells and negroid Hb AA cells. 2. The erythrocyte concentration of sodium was 40% higher and potassium 10% lower in the Hb SS than in the Hb AA cells. The cell water expressed as % weight of cell (corrected for trapped plasma) was identical for both cell types. 3. Normal Caucasian erythrocytes with Hb AA contained 40–50% less sodium but about the same potassium concentration as negroid Hb AA cells. 4. Potassium efflux into buffered iso-osmotic sucrose medium was much faster in Hb SS than in negroid Hb AA cells; ouabain-sensitive active sodium transport was twice as fast in the sickle-cell erythrocytes. Passive sodium efflux of erythrocytes suspended in a physiological medium was similarly faster in Hb SS cells. 5. Under the conditions of the experiments not less than 85% of the Hb SS erythrocytes appeared biconcave. Electron-microscopic examination of ultra-thin sections of Hb SS cells revealed marked discontinuities in the membrane. This suggests definite membrane alterations, which have probably resulted from the sickling-unsickling cycles occurring during the life-span of the cells. 6. It is suggested that the enhanced active sodium transport in the Hb SS erythrocyte is secondary to the augmented passive cation efflux, which in turn results from the leakiness of the erythrocyte membrane produced by the sickling-unsickling process.

Light and Electron Microscopic Examination of the Parasitic Dinoflagellate Haplozoon

2000 ◽  
Vol 6 (S2) ◽  
pp. 884-885
Author(s):  
Stephen C. Landers
Keyword(s):  
Gulf Coast ◽  
Light And Electron Microscopy ◽  
Electron Microscopic Examination ◽  
The United States ◽  
Intestinal Wall ◽  
State University ◽  
University Campus ◽  
Electron Microscopic ◽  
Daughter Cells ◽  
A Chain

The parasitic dinoflagellate Haplozoon is found in the intestine of marine polychaetes. It is composed of a chain of cells that hang from the intestinal wall into the lumen, and releases daughter cells from the posterior end of the chain which leave the host and reinfect other polychaetes. Few studies exist in the recent literature regarding Haplozoon and it has not been reported from the Gulf Coast of the United States. This study reports the genus Haplozoon from the maldanid polychaete Axiothella mucosa in St. Andrew Bay, Florida, and examines the structure of the organism by light and electron microscopy.Axiothella mucosa was collected in St. Andrew Bay, Florida and maintained in seawater at the Troy State University campus. Haplozoon spp. was prepared for whole mounts by smearing minced setigers of the worms onto a slide with a drop of seawater.

Electron microscopic analysis of objects in light microscopic sections

1978 ◽  
Vol 36 (2) ◽  
pp. 80-81
Author(s):  
Arvid B. Maunsbach
Keyword(s):  
Electron Microscopy ◽  
Light And Electron Microscopy ◽  
Microscopic Analysis ◽  
Electron Microscopic Level ◽  
Semithin Section ◽  
Cover Glass ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Glass Slides ◽  
Ultrathin Sections

Structural studies in experimental biology or in pathology are frequently extended from the light to the electron microscopic level. This is often done by cutting both semithin (about 1 μm) and thin sections from the same tissue block after embedding for electron microscopy. However, in many studies it would be of great value to analyse the same structure both by light and electron microscopy, i.e. to be able to study by electron microscopy an object which is first detected by light microscopy in a semithin section. To achieve this, a method has been developed by which ultrathin sections are cut directly from the semithin section containing the object of interest.Semithin sections, about 1 μ in thickness, are cut from Epon or Vestopal embedded tissue. The sections are placed on ordinary glass slides and stained with toluidine blue. The sections are studied in the light microscope without a cover glass or mounted in water.

Effect of Streptozotocin-Induced Diabetes on Estradiol-Stimulated Changes in the Ultrastructure of the Rat Endometrium

1981 ◽  
Vol 39 ◽  
pp. 532-533
Author(s):  
G.T. Frederick ◽  
R.M. Gardner ◽  
J.M. Kirkland ◽  
G.M. Stancel
Keyword(s):  
Electron Microscopic Examination ◽  
Sprague Dawley ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Whole Cells ◽  
Glucose Levels ◽  
Blood Glucose Levels ◽  
Sprague Dawley Rats ◽  
Uterine Growth ◽  
Streptozotocin Induced Diabetes

Estradiol (E2)-stimulated uterine growth has been well characterized both biochemically and morphologically. Recent studies have shown that insulin plays an important role in the regulation of cellular development. This study examines the effect of streptozotocin-induced diabetes on E2-stimulated changes in the ultrastructure of the endometrium of the rat uterus.Sprague-Dawley rats were ovariectomized at 21 days of age. Diabetes, defined as blood glucose levels greater than 300mg%, was induced in half of these animals by the injection of 85mg streptozotocin/kg body weight.The remaining animals were classified as normal. Animals from both groups were injected with either 0.9% saline or 4μg E2/100gm body weight. At 18, 24, 36 and 48 hours post-injection of E2, uterine segments were collected from 8-10 animals in each experimental group and processed for electron microscopic examination. Uterine segments from saline-injected animals served as controls. Data summarized describe observations made from thin-sections and have not been extrapolated for whole cells.

Electron microscopic evidence for a double hair-like nap appearing at low frequency on Bacillus anthracis Sterne spores

1969 ◽  
Vol 15 (10) ◽  
pp. 1247-1248 ◽  
Author(s):  
M. J. Kramer ◽  
Ivan L. Roth
Keyword(s):  
Bacillus Anthracis ◽  
Microscopic Examination ◽  
Low Frequency ◽  
Electron Microscopic Examination ◽  
Uranyl Acetate ◽  
Spore Coat ◽  
Lead Citrate ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Very Low Frequency

Electron microscopic examination of thin sections of Bacillus anthracis Sterne spores triply poststained with KMnO4, uranyl acetate, and lead citrate has indicated an unusual morphological variant. These spores are seen at very low frequency and have, in addition to the hair-like nap normally associated with the exosporium a second hairy layer which appears to originate in the spore coat complex.

The influence of aluminium on iron oxides: IX. Dissolution of Al-goethites in 6 M HCl

Clay Minerals ◽  
1984 ◽  
Vol 19 (1) ◽  
pp. 9-19 ◽  
Author(s):  
U. Schwertmann
Keyword(s):  
Crystal Growth ◽  
Surface Area ◽  
Electron Microscopic Examination ◽  
Separate Experiment ◽  
Dissolution Time ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Al Substitution ◽  
Crystallite Dimension ◽  
Oriented Parallel

AbstractThe rate of dissolution of synthetic Al-goethites with 0–10 mole% Al in 6 m HCl at 24°C decreases markedly with increasing extent of Al substitution. Most of the dissolution-time curves are S-shaped, suggesting some increase in surface area during the initial stages of dissolution. Based on electron microscopic examination, the increase in surface area is explained by preferential dissolution of less ordered ‘interdomainic’ zones and the consequent production of isolated ‘domains’. Thin sections of crystals provided some support for defect zones oriented parallel to the crystallographic c-direction. Stepwise multiple correlation analysis employing various properties of the goethites was used to investigate the large variation in half-dissolution time (1–96 h). 94% of the variation could be explained by the variation of the mean crystallite dimension perpendicular to the planes (110) and (111) (MCD110, MCD111). Inclusion of Al-substitution increased R2 to only 97%. As shown in a separate experiment, increasing Al concentration in the system retarded crystal growth. It is therefore believed that Al affects the dissolution rate of goethites not directly, but indirectly, by influencing crystal growth rate which, in turn, affects crystal size and order as measured by MCD110 and MCD111.

scholarly journals Use of anti-horseradish peroxidase antibody-gold complex in the ABC technique.

1991 ◽  
Vol 39 (6) ◽  
pp. 863-869 ◽  
Author(s):  
B Gee ◽  
M J Warhol ◽  
J Roth
Keyword(s):  
Horseradish Peroxidase ◽  
Polyclonal Antibodies ◽  
Light And Electron Microscopy ◽  
Gold Complex ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Endogenous Peroxidase ◽  
Pre Treatment ◽  
Lowicryl K4m ◽  
Antigen Antibody

We report a modification of the avidin-biotin-peroxidase complex (ABC) technique for the light and electron microscopic detection of antigens in tissue sections. An immunological approach was used instead of the DAB reaction to reveal ABC bound to antigen-antibody complexes. Affinity-purified polyclonal antibodies against horseradish peroxidase were complexed to particles of colloidal gold and applied for reaction with the horseradish peroxidase molecules of the ABC. For light microscopic immunolabeling, the signal produced by the anti-horseradish peroxidase antibody-gold complex required silver intensification. The ABC immunogold reaction as compared with the standard ABC technique, in particular with silver intensification of the DAB reaction product, provided superior resolution in paraffin sections. Furthermore, section pre-treatment to block endogenous peroxidase activity could be omitted and no potentially hazardous substrate was used. The ABC immunogold reaction was successfully applied for electron microscopic immunolabeling on Lowicryl K4M thin sections. We propose that the ABC immunogold reaction is a useful alternative to the standard ABC technique and can be equally well applied to light and electron microscopy.

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