Ultrastructural localization of Sm28 GST protective antigen in Schistosoma mansoni adult worms

Parasitology ◽  
1996 ◽  
Vol 113 (4) ◽  
pp. 377-391 ◽  
Author(s):  
J. L. Liu ◽  
J. Fontaine ◽  
A. Capron ◽  
J. M. Grzych
Keyword(s):  
Schistosoma Mansoni ◽  
Protective Antigen ◽  
Ultrastructural Localization ◽  
Ultrastructural Level ◽  
Staining Intensity ◽  
Wide Distribution ◽  
Published Data ◽  
Glutathione S Transferase ◽  
Sustentacular Cells ◽  
Germinal Cells

SUMMARYThe localization of the 28 kDa Schistosoma mansoni glutathione S-transferase (Sm28 GST) has been investigated using immunohistochemistry and electron microscopy and the results compared with previously published data. This study confirms the wide distribution of this antigen in the parasite. In male and female worms, Sm28 GST is localized in the tegument, the parenchyma, the oesophageal epithelium and in genital organs. Sm28 GST was clearly detected in germinal and sustentacular cells. The decrease of staining intensity during the differentiation of germinal cells suggests a down-regulated expression of the molecule. At the ultrastructural level, this antigen was abundant in nuclei and less present in the cytoplasm. The marked heterogeneity observed in the staining of individual worms indicates that Sm28 GST seems to be closely associated with the parasite’s metabolism. The results are discussed in relation to the biological and protective functions of the protein.

Schistosoma mansoni: the sustentacular cells of the testes

Parasitology ◽  
1981 ◽  
Vol 82 (1) ◽  
pp. 125-130 ◽  
Author(s):  
O. A. Otubanjo
Keyword(s):  
Endoplasmic Reticulum ◽  
Schistosoma Mansoni ◽  
Potassium Permanganate ◽  
Basal Lamina ◽  
Cell Types ◽  
Principal Cell ◽  
Sustentacular Cells ◽  
Glycogen Particles ◽  
Germinal Cells

SUMMARYStudies on the multiple compact testes of Schistosoma mansoni have revealed 2 principal cell types: germinal and non-germinal cells. Two morphological forms representing different stages in the cytomorphosis of the sustentacular cells have been considered. The polymorphic sustentacular cells which are conspicuously stained with potassium permanganate contain abundant ribosomal endoplasmic reticulum, glycogen particles and ribosomal masses in comparison with the germinal component of the testes. Golgi complexes with associated secretory bodies and vacuoles are present in the cells. The cytoplasm of the sustentacular cells interdigitate between other cells in the testes. Based on their morphology, nutritive, supportive and phagocytic functions have been attributed to the sustentacular cells. The possible recycling of nutrients by the cells is discussed. It is suggested that the cells regulate the production of spermatozoa from a nutritive viewpoint and also that the translocation of nutrients from the parenchyma to the testes is facilitated by these cells whose cytoplasm extends directly adjacent to the basal lamina.

scholarly journals Ultrastructural localization of cyclic GMP and cyclic AMP in rat striatum.

1981 ◽  
Vol 91 (1) ◽  
pp. 287-292 ◽  
Author(s):  
M A Ariano ◽  
A I Matus
Keyword(s):  
Cyclic Amp ◽  
Glial Cells ◽  
Cyclic Gmp ◽  
Second Messengers ◽  
Ultrastructural Localization ◽  
Ultrastructural Level ◽  
Staining Intensity ◽  
Synaptic Function ◽  
Rat Striatum ◽  
Synaptic Localization

The subcellular localization of cyclic GMP and cyclic AMP in the rat caudate-putamen has been studied using horseradish peroxidase immunocytochemistry. Both of the putative neurotransmitter second messengers were visualized in neurons and glial cells at light microscopic resolutions, but not all cells of either category gave detectable staining. This was confirmed at the ultrastructural level where both stained and unstained elements of the same cell type were found within the same field. A striking variation was seen in cyclic nucleotide staining intensity within individual neural and glial cells. Both of the cyclic nucleotides were detected within postsynaptic terminal boutons and within astroglial processes. Cyclic GMP postsynaptic staining was stronger than glial staining, whereas the localization pattern was reversed for cyclic AMP. The synaptic localization of cyclic AMP and cyclic GMP immunoreactivity adds support to the idea that these compounds have an influential role in synaptic function within the striatum.

scholarly journals Application of the FITC-anti-FITC-gold system to ultrastructural localization of antigens.

1991 ◽  
Vol 39 (12) ◽  
pp. 1725-1728 ◽  
Author(s):  
G J van Dam ◽  
B J Bogitsh ◽  
J A Fransen ◽  
D Kornelis ◽  
R J van Zeyl ◽  
...  
Keyword(s):  
Schistosoma Mansoni ◽  
Fluorescein Isothiocyanate ◽  
High Specificity ◽  
Ultrastructural Localization ◽  
Good Alternative ◽  
Ultrastructural Level ◽  
Electron Microscopic ◽  
Protein Conjugates ◽  
Specificity And Sensitivity ◽  
Microscopic Studies

We report the application of a fluorescein isothiocyanate (FITC)-anti-FITC method to localize antigens at the ultrastructural level. In the systems studied, the anti-FITC-based detection method displays high specificity and sensitivity. These observations, combined with ease of production and with availability of FITC-protein conjugates, suggest that the FITC-anti-FITC method is a good alternative to presently used methods and is widely applicable to immunochemical and immunocytochemical procedures. The same preparation and protocol can be used for light and electron microscopic studies, thereby reducing possible artifacts introduced if different procedures are used. In the present study, two systems were used to test the method. One system used an FITC-labeled monoclonal antibody (MAb) to schistosome circulating cathodic antigen. In this system, the label was detected in the gut of adult Schistosoma mansoni by an anti-FITC MAb conjugated to 10-nm gold particles. The second system used human IgM antibodies pooled from patients infected with Schistosoma mansoni. In this system detection was accomplished using an anti-human IgM-FITC conjugate followed by the anti-FITC-Au antibody conjugate.

Ultrastructural Localization of Antigens by Capillary Action Immunocytochemistry

1996 ◽  
Vol 54 ◽  
pp. 40-41
Author(s):  
László G. Kömüves
Keyword(s):  
San Francisco ◽  
Colloidal Gold ◽  
Specific Binding ◽  
Ultrastructural Localization ◽  
Ultrastructural Level ◽  
Uranyl Acetate ◽  
Adhesive Tape ◽  
Distilled Water ◽  
Capillary Action ◽  
Rabbit Igg

Light microscopic immunohistochemistry based on the principle of capillary action staining is a widely used method to localize antigens. Capillary action immunostaining, however, has not been tested or applied to detect antigens at the ultrastructural level. The aim of this work was to establish a capillary action staining method for localization of intracellular antigens, using colloidal gold probes.Post-embedding capillary action immunocytochemistry was used to detect maternal IgG in the small intestine of newborn suckling piglets. Pieces of the jejunum of newborn piglets suckled for 12 h were fixed and embedded into LR White resin. Sections on nickel grids were secured on a capillary action glass slide (100 μm wide capillary gap, Bio-Tek Solutions, Santa Barbara CA, distributed by CMS, Houston, TX) by double sided adhesive tape. Immunolabeling was performed by applying reagents over the grids using capillary action and removing reagents by blotting on filter paper. Reagents for capillary action staining were from Biomeda (Foster City, CA). The following steps were performed: 1) wet the surface of the sections with automation buffer twice, 5 min each; 2) block non-specific binding sites with tissue conditioner, 10 min; 3) apply first antibody (affinity-purified rabbit anti-porcine IgG, Sigma Chem. Co., St. Louis, MO), diluted in probe diluent, 1 hour; 4) wash with automation buffer three times, 5 min each; 5) apply gold probe (goat anti-rabbit IgG conjugated to 10 nm colloidal gold, Zymed Laboratories, South San Francisco, CA) diluted in probe diluent, 30 min; 6) wash with automation buffer three times, 5 min each; 7) post-fix with 5% glutaraldehyde in PBS for 10 min; 8) wash with PBS twice, 5 min each; 9) contrast with 1% OSO4 in PBS for 15 min; 10) wash with PBS followed by distilled water for5 min each; 11) stain with 2% uranyl acetate for 10 min; 12) stain with lead citrate for 2 min; 13) wash with distilled water three times, 1 min each. The glass slides were separated, and the grids were air-dried, then removed from the adhesive tape. The following controls were used to ensure the specificity of labeling: i) omission of the first antibody; ii) normal rabbit IgG in lieu of first antibody; iii) rabbit anti-porcine IgG absorbed with porcine IgG.

Immunohistochemical and ultrastructural localization of Schistosoma mansoni soluble egg antigens processed by the infected host

Parasitology ◽  
1996 ◽  
Vol 112 (6) ◽  
pp. 537-543 ◽  
Author(s):  
J. J. P. M. Bogers ◽  
H. A. M. Nibbeling ◽  
A. M. Deelder ◽  
E. A. E. Van Marck
Keyword(s):  
Schistosoma Mansoni ◽  
Indirect Evidence ◽  
Ultrastructural Localization ◽  
Carbohydrate Epitopes ◽  
Eosinophilic Granulocytes ◽  
Antigenic Material ◽  
Egg Shells ◽  
Secondary Lysosomes ◽  
Antigen Antibody ◽  
Egg Antigen

SUMMARYThe detection of egg-derived antigens in the serum and urine of Schistosoma mansoni-infected individuals and experimental animals would provide an alternative method to assess the tissue egg burden. The detected levels are, however, not only a function of the amounts of antigen produced, but also of the processing or clearance by the host. In the present study the immunolocalization pattern of antigens using 2 recently described monoclonal antibodies to repetitive carbohydrate epitopes of S. mansoni soluble egg antigen (114–5B1–A and 114–4D12–A) in various organs of the host was investigated. In the liver strong immunoreactivity could be detected around the entrapped eggs and in egg-shells, as well as in Kupffer cells accumulating both antigen and schistosomal pigment. In the spleen, immunohistochemistry revealed antigen in the plasma as well as in secondary lysosomes of macrophages. Strong labelling was found in the vesicles of the eosinophilic granulocytes: indirect evidence perhaps for the presence of antigen–antibody complexes. In conclusion, the secreted egg antigens were sequestered in the reticulo-endothelial macrophages of the liver and the spleen as already partly described for worm-derived antigens. The presence of large quantities of antigenic material in the spleen could suggest an important role of this organ in the clearance of antigen and might even provide an additional explanation for the hepatosplenomegaly mainly present in S. mansoni-infected children.

scholarly journals The organelles of the trans domain of the cell. Ultrastructural localization of sialoglycoconjugates using Limax flavus agglutinin.

1986 ◽  
Vol 34 (8) ◽  
pp. 1069-1077 ◽  
Author(s):  
K Hedman ◽  
I Pastan ◽  
M C Willingham
Keyword(s):  
Plasma Membrane ◽  
Ultrastructural Localization ◽  
Ultrastructural Level ◽  
Coated Pits ◽  
Golgi Stack ◽  
Acetylneuraminic Acid ◽  
Secretory Glycoproteins ◽  
Double Label ◽  
Endocytic Vesicles ◽  
Exocytic Pathway

The subcellular distribution of sialic acid was determined at the ultrastructural level using Limax flavus agglutinin (LFA). This lectin, which is specific for N-acetylneuraminic acid and N-glycolylneuraminic acid, was covalently conjugated to horseradish peroxidase (HRP). The conjugates (LFA-HRP) were applied to aldehyde-fixed, saponin-permeabilized 3T3 cells in pre-embedding labeling electron microscopy. Peroxidase label was detected in a patchy distribution at the cell surface, and in plasma-membrane-coated pits, endocytic vesicles (receptosomes), multivesicular bodies, and lysosomes. Smooth-surfaced tubular and vesicular structures, similar to those that participate in membrane recycling, were labeled. In the Golgi complex, more than half of the cisternae contained label--typically only one cisterna on the cis side was unlabeled. Heavily labeled structures of the trans Golgi included a reticular membranous system with coated regions--50-80 nm diameter vesicular or pit-like profiles and larger coated vacuoles. Smooth 200-300 nm vacuoles were labeled on the trans side of the Golgi stack. Similar structures have been previously shown to participate in the exocytosis of plasma membrane and secretory glycoproteins from the Golgi stacks. These findings identify those intracellular organelles that are functionally at the level of, or distal to, the sialyltransferase-containing membranes of the Golgi, and distinguish them from the pre-Golgi membranous structures. The LFA-HRP conjugate is an indicator for this functional trans domain of the cell, and should be applicable for ultrastructural double-label experiments as a cis versus trans marker of the exocytic pathway.

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