scholarly journals Antibodies to the Golgi complex and the rough endoplasmic reticulum.

1982 ◽  
Vol 92 (1) ◽  
pp. 92-107 ◽  
Author(s):  
D Louvard ◽  
H Reggio ◽  
G Warren
Keyword(s):  
Endoplasmic Reticulum ◽  
Rough Endoplasmic Reticulum ◽  
Golgi Complex ◽  
Rat Kidney ◽  
Apparent Molecular Weight ◽  
Fluorescent Labeling ◽  
Central Feature ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Culture Cells

Rabbits were immunized with membrane fractions from either the Golgi complex or the rough endoplasmic reticulum (RER) by injection into the popliteal lymph nodes. The antisera were then tested by indirect immunofluorescence on tissue culture cells or frozen, thin sections of tissue. There were may unwanted antibodies to cell components other than the RER or the Golgi complex, and these were removed by suitable absorption steps. These steps were carried out until the pattern of fluorescent labeling was that expected for the Golgi complex or RER. Electron microscopic studies, using immunoperoxidase labeling of normal rat kidney (NRK) cells, showed that the anti-Golgi antibodies labeled the stacks of flattened cisternae that comprise the central feature of the Golgi complex, many of the smooth vesicles around the stacks, and a few coated vesicles. These antibodies were directed, almost entirely, against a single polypeptide with an apparent molecular weight of 135,000. The endoplasmic reticulum (ER) in NRK cells is an extensive, reticular network that pervades the entire cell cytoplasm and includes the nuclear membrane. The anit-RER antibodies labeled this structure alone at the light and electron microscopic levels. They were largely directed against four polypeptides with apparent molecular weights of 29,000, 58,000, 66,000, and 91,000. Some examples are presented, using immunofluorescence microscopy, where these antibodies have been used to study the Golgi complex and RER under a variety of physiological and experimental condition . For biochemical studies, these antibodies should prove useful in identifying the origin of isolated membranes, particularly those from organelles such as the Golgi complex, which tend to lose their characteristic morphology during isolation.

OBSERVATIONS ON THE LOCALIZATION OF RECENTLY SYNTHESIZED CATECHOLAMINES IN CHROMAFFIN CELLS AFTER THE INJECTION OF l-[2,5,6-3H]DOPA

1976 ◽  
Vol 69 (1) ◽  
pp. 139-NP ◽  
Author(s):  
R. E. COUPLAND ◽  
S. KOBAYASHI ◽  
CHRISTINE KENT
Keyword(s):  
Endoplasmic Reticulum ◽  
Rough Endoplasmic Reticulum ◽  
Golgi Complex ◽  
Chromaffin Cells ◽  
Intracellular Localization ◽  
Chromaffin Granules ◽  
Electron Microscopic ◽  
Electron Microscopic Autoradiography ◽  
Metabolic Products ◽  
Noradrenaline And Adrenaline

SUMMARY The fate of l-[2,5,6-3H]DOPA, and the intracellular localization of its metabolic products dopamine, noradrenaline and adrenaline, have been determined by the simultaneous use of assay techniques following separation of amines by chromatography and light and electron microscopic autoradiography. During the first 24 h after i.v. or i.p. injection of [3H]DOPA, synthesis of the above catecholamines occurred. Throughout this time the labelled amines were associated with chromaffin granules or immediately adjacent cytosol and not with either the Golgi complex or rough endoplasmic reticulum. Labelling of chromaffin granules occurred simultaneously throughout the cell and there was no evidence of regions containing recently labelled granules and others containing previously charged (older) granules. Adrenaline-storing cells took up [3H]DOPA and its products more rapidly and lost recently synthesized adrenaline more rapidly than noradrenaline-storing cells took up and stored their equivalent amines. This was in keeping with a more rapid turnover of catecholamines in adrenaline-storing elements.

scholarly journals Immunoreactive kallikrein localization in the rat kidney: an immuno-electron-microscopic study.

1984 ◽  
Vol 32 (1) ◽  
pp. 117-121 ◽  
Author(s):  
C D Figueroa ◽  
I Caorsi ◽  
J Subiabre ◽  
C P Vío
Keyword(s):  
Endoplasmic Reticulum ◽  
Electron Microscope ◽  
Golgi Complex ◽  
Plasma Membranes ◽  
Rat Kidney ◽  
Immunocytochemical Staining ◽  
Cell Type ◽  
Tubule Cell ◽  
Electron Microscopic ◽  
Pap Method

The cellular and subcellular localization of immunoreactive kallikrein was studied in the rat kidney using the peroxidase-antiperoxidase (PAP) method for the electron microscope. The effect of various tissue-processing protocols on ultrastructural preservation and immunocytochemical staining was evaluated by fixing kidneys with four different mixtures. The tissues were immunostained and further stained with OsO4 or silver methenamine. The best ultrastructural and immunocytochemical staining was obtained with Zamboni's-glutaraldehyde fixative. The kallikrein-immunoreactive cell type was identified, according to its localization and ultrastructural features, as the connecting-tubule cell. Immunoreactive kallikrein was concentrated mainly in the upper one-third of the cell and at both sides of the nuclei, and to a less extent was associated with the plasma membranes and basolateral infoldings. The immunoreactivity was related to free polyribosomes, the rough endoplasmic reticulum (RER), and the Golgi complex, suggesting that kallikrein is actively synthesized in this particular type of cell.

scholarly journals Glycosyltransferase activities in Golgi complex and endoplasmic reticulum fractions isolated from African trypanosomes.

1984 ◽  
Vol 99 (2) ◽  
pp. 569-577 ◽  
Author(s):  
D J Grab ◽  
S Ito ◽  
U A Kara ◽  
L Rovis
Keyword(s):  
Endoplasmic Reticulum ◽  
Rough Endoplasmic Reticulum ◽  
Golgi Complex ◽  
Smooth Endoplasmic Reticulum ◽  
Surface Glycoprotein ◽  
Relative Distribution ◽  
African Trypanosomes ◽  
Independent Form ◽  
Variable Surface ◽  
Dependent Form

Highly enriched Golgi complex and endoplasmic reticulum fractions were isolated from total microsomes obtained from Trypanosoma brucei, Trypanosoma congolense, and Trypanosoma vivax, and tested for glycosyltransferase activity. Purity of the fractions was assessed by electron microscopy as well as by biochemical analysis. The relative distribution of all the glycosyltransferases was remarkably similar for the three species of African trypanosomes studied. The Golgi complex fraction contained most of the galactosyltransferase activity followed by the smooth and rough endoplasmic reticulum fractions. The dolichol-dependent mannosyltransferase activities were highest for the rough endoplasmic reticulum, lower for the smooth endoplasmic reticulum, and lowest for the Golgi complex. Although the dolichol-independent form of N-acetylglucosaminyltransferase was essentially similar in all the fractions, the dolichol-dependent form of this enzyme was much higher in the endoplasmic reticulum fractions than in the Golgi complex fraction. Inhibition of this latter activity in the smooth endoplasmic reticulum fraction by tunicamycin A1 suggests that core glycosylation of the variable surface glycoprotein may occur in this organelle and not in the rough endoplasmic reticulum as previously assumed.

scholarly journals Expression of Enamel Proteins in Rough Endoplasmic Reticulum and Golgi Complex in Human Dental Germs

2017 ◽  
Vol 35 (2) ◽  
pp. 435-441
Author(s):  
Francisco Javier Gutiérrez-Cantú ◽  
Alma Lilián Guerrero-Barrera ◽  
Wulfrano Sánchez Meraz ◽  
Amaury de Jesús Pozos-Guillen ◽  
Héctor Flores-Reyes ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Rough Endoplasmic Reticulum ◽  
Golgi Complex ◽  
Enamel Proteins

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.

scholarly journals Immunocytochemical localization of D-amino acid oxidase in the central clear matrix of rat kidney peroxisomes.

1986 ◽  
Vol 34 (12) ◽  
pp. 1709-1718 ◽  
Author(s):  
N Usuda ◽  
S Yokota ◽  
T Hashimoto ◽  
T Nagata
Keyword(s):  
Amino Acid ◽  
Proximal Tubule ◽  
Rat Kidney ◽  
Protein A ◽  
Immunoblot Analysis ◽  
Acid Oxidase ◽  
Electron Microscopic ◽  
Amino Acid Oxidase ◽  
Thin Sections ◽  
Gold Particles

Light and electron microscopic localizations of D-amino acid oxidase (DAO) in rat kidney was investigated using immunoenzyme and protein A-gold techniques. The enzyme was purified from rat kidney homogenate and its antibody was raised in rabbits. By Ouchterlony double-diffusion analysis and immunoblot analysis with anti-(rat kidney DAO) immunoglobulin, the antibody was confirmed to be monospecific. The tissue sections (200 micron thick) of fixed rat kidney were embedded in Epon or Lowicryl K4M. Semi-thin sections were stained for DAO by the immunoenzyme technique after removal of epoxy resin for LM, and ultra-thin sections of Lowicryl-embedded material were labeled for DAO by the protein A-gold technique for EM. By LM, fine cytoplasmic granules of proximal tubule were stained exclusively. Among three segments of proximal tubules, and S2 and S3 segments were heavily stained but the S1 segment only weakly so. By EM, gold particles indicating the antigenic sites for DAO were exclusively confined to peroxisomes. Within peroxisomes, the gold particles were localized in the central clear matrix but not in the peripheral tubular substructures. The results indicate that D-amino acid oxidase in rat kidney is present exclusively in peroxisomes in the proximal tubule and that within peroxisomes it is found only in central clear matrix and not in the peripheral tubular substructures.

scholarly journals ELECTRON MICROSCOPY OF THE BURSA OF FABRICIUS OF THE EMBRYONIC CHICK WITH PARTICULAR REFERENCE TO THE LYMPHO-EPITHELIAL NODULES

1962 ◽  
Vol 13 (1) ◽  
pp. 127-146 ◽  
Author(s):  
G. Adolph Ackerman
Keyword(s):  
Endoplasmic Reticulum ◽  
Epithelial Cells ◽  
Embryonic Development ◽  
Golgi Complex ◽  
Bursa Of Fabricius ◽  
Cortical Region ◽  
Electron Microscopic ◽  
Submicroscopic Structure ◽  
Microscopic Studies ◽  
Pass Through

Electron microscopic studies of the bursa of Fabricius during the 15th and 16th day of embryonic development in the chick have shown the following findings in the submicroscopic structure of the cellular elements of the lympho-epithelial follicles. In the medulla, basal endodermal epithelial cells undergo mitosis and differentiation into lymphoblasts. During this transformation, there is a reduction in the amount of rough endoplasmic reticulum, an increase in the number or ribosomes, and frequently an enlargement of the Golgi complex. As lymphoblasts differentiate into medium lymphocytes there is a loss of endoplasmic reticulum, a reduction in the number of ribosomes and in the size of the Golgi complex, as well as a decrease in the number and size of mitochondria and in the size of the cell and nucleus. Cytoplasmic processes of reticular-epithelial cells extend between proliferating lymphocytic cells. Desmosomes connect stellate reticular-epithelial and basal epithelial cells but are not present in lymphocytic cells. Nuclear blebbing and vesiculation are frequently observed in the various cell forms of the developing lympho-epithelial nodules. Although lymphocytes and lymphocytopoietic activities in the cortex are sparse during this stage of embryonic development of the bursa, transitional forms between mesenchymal cells and lymphoblasts have been encountered. In addition, lymphoblasts and/or undifferentiated epithelial cells occasionally may pass through the basement membrane from the medulla into the cortical region of the developing nodule. That lymphocytes in the bursa of Fabricius originate from both endodermal and mesodermal derivatives during embryonic development appears to be consistent with both light and electron microscopic observations.

scholarly journals Phosphatidylinositol kinase and diphosphoinositide kinase of rat kidney cortex: properties and subcellular localization

1976 ◽  
Vol 160 (1) ◽  
pp. 97-105 ◽  
Author(s):  
P H Cooper ◽  
J N Hawthorne
Keyword(s):  
Endoplasmic Reticulum ◽  
Golgi Complex ◽  
Brush Border ◽  
Rat Kidney ◽  
Subcellular Fractionation ◽  
Kidney Cortex ◽  
Marker Enzymes ◽  
Rat Kidney Cortex ◽  
Phosphatidylinositol Kinase ◽  
Golgi Membranes

The properties of phosphatidylinositol kinase and diphosphoinositide kinase from rat kidney cortex were studied. The enzymes were completely Mg2+-dependent. Cutscum detergent activated phosphatidylinositol kinase, but diphosphoinositide kinase was inhibited by all detergents tested. The pH optima were 7.7 for phosphatidylinositol kinase and 6.5 for diphosphoinositide kinase. On subcellular fractionation of kidney-cortex homogenates by differential centriflgation, the distribution of phosphatidylinositol kinase resembled that of the marker enzymes for brush-border, endoplasmic-reticulum and Golgi membranes. Diphosphoinositide kinase distribution resembled that of thiamin pyrophosphatase (assayed in the absence of ATP), diphosphoinositide phosphatase and triphosphoinositide phosphatase. Activities of both kinases were low in purified brush-border fragments. Diphosphoinositide kinase is probably localized in the Golgi complex.

Electron microscopic observations of Toxoplasma ‘Nicolle et Manceaux’ in thin sections of tissue cultures

Parasitology ◽  
1957 ◽  
Vol 47 (1-2) ◽  
pp. 66-69 ◽  
Author(s):  
H. Meyer ◽  
I. De Andrade Mendonça
Keyword(s):  
Endoplasmic Reticulum ◽  
Research Council ◽  
National Research Council ◽  
Cost Of Reproduction ◽  
Tissue Cultures ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Double Membrane ◽  
Opposite Pole ◽  
The Cost

Toxoplasma ‘Nicolle et Manceaux’ has been examined in the electron microscope in thin sections of infected tissue cultures. The intracellular forms, single or in rosette formations, show that the parasite has no flagella, cilia or other organelles for locomotion. It is covered by a double membrane which is especially well denned at the two poles. A ring-like formation, often slightly protruding, can be seen at one end of the parasite, and in connexion with this, long, homogeneous, dark-stained inclusions of still unknown nature. At the opposite pole a distinct opening has been found.Mitochondria and strands of an endoplasmic reticulum are also present in the cytoplasm and a fine striation in the region of the periplast. No dividing forms have been observed. Near the nucleus, in the region of what may correspond to the centrosome and the Golgi apparatus, a few extremely fine granules and a spiral-like formation of fine striae or tubules have been observed.This work has been partly supported by the National Research Council of Brazil. The cost of reproduction of figures was defrayed by the Institute de Biofísica da Universidade do Brasil.

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