scholarly journals HISTOCHEMICAL AND ULTRASTRUCTURAL DEMONSTRATION OF γ-GLUTAMYL TRANSPEPTIDASE ACTIVITY

1969 ◽  
Vol 17 (8) ◽  
pp. 517-526 ◽  
Author(s):  
ALEXANDER M. RUTENBURG ◽  
HWAKYU KIM ◽  
JEROME W. FISCHBEIN ◽  
JACOB S. HANKER ◽  
HANNAH L. WASSERKRUG ◽  
...  
Keyword(s):  
Azo Dye ◽  
Histochemical Demonstration ◽  
Zymogen Granules ◽  
Optimum Conditions ◽  
Histochemical Reaction ◽  
Copper Chelate ◽  
Electron Microscopic ◽  
Apical Portion ◽  
Glutamyl Transpeptidase ◽  
Electron Microscopic Localization

A simultaneous coupling azo dye method for the histochemical demonstration of γ-glutamyl transpeptidase activity using the new substrate γ-glutamyl-4-methoxy-2-naphthylamide has been described. The method appears superior to previously reported methods for γ-glutamyl transpeptidase activity and can easily be modified for the electron microscopic localization of the enzyme by bridging osmium to the copper chelate of the azo dye via thiocarbohydrazide. The optimum conditions for the histochemical reaction were developed and the distribution of enzymatic activity in the tissues of the rat is described for light microscopy and with rat pancreas for electron microscopy. The electron-opaque deposits were seen in the endoplasmic reticulum in the vicinity of the zymogen granules in the apical portion of the acinar cell.

scholarly journals The histochemical demonstration of gamma-glutamyl transpeptidase activity in different populations of rat liver during azo dye carcinogenesis.

1982 ◽  
Vol 30 (4) ◽  
pp. 312-316 ◽  
Author(s):  
R Daoust
Keyword(s):  
Bile Duct ◽  
Azo Dye ◽  
Histochemical Demonstration ◽  
Cell Populations ◽  
Liver Lesions ◽  
Cell Type ◽  
Gamma Glutamyl Transpeptidase ◽  
Gamma Glutamyl ◽  
Different Populations ◽  
Glutamyl Transpeptidase

To better assess the reliability of gamma-glutamyl transpeptidase (gamma-GTase) as a marker of preneoplastic liver lesions and hepatomas, the gamma-GTase activity of different cell populations was examined in liver sections from rats fed 4-dimethylaminoazobenzene. The results indicated that the biliary ductular cells in trabeculae of cirrhotic livers may exhibit appreciable gamma-GTase activity in addition to that shown by islands of regenerating parenchyma. At later stages of azo dye carcinogenesis, the epithelial cells of bile duct cysts and cholangiomas, as well as those of hepatomas, gave positive reactions for gamma-GTase. Thus biochemical data on liver gamma-GTase in different models of hepatocarcinogenesis cannot be translated directly in terms of alterations in a particular cell type unless such interpretation is justified by parallel histochemical investigations.

scholarly journals MEMBRANEOUS ULTRASTRUCTURAL DEMONSTRATION OF AMINOPEPTIDASE AND γ-GLUTAMYL TRANSPEPTIDASE ACTIVITIES WITH A NEW DIAZONIUM SALT THAT YIELDS A LIPOPHOBIC, OSMIOPHILIC AZO DYE

1970 ◽  
Vol 18 (8) ◽  
pp. 542-551 ◽  
Author(s):  
ARNOLD M. SELIGMAN ◽  
HANNAH L. WASSERKRUG ◽  
ROBERT E. PLAPINGER ◽  
TAKASHI SEITO ◽  
JACOB S. HANKER
Keyword(s):  
Light Microscopy ◽  
Enzymatic Activity ◽  
Azo Dyes ◽  
Azo Dye ◽  
Diazonium Salt ◽  
Aminopeptidase Activity ◽  
Electron Microscopic ◽  
New Methods ◽  
Droplet Distribution ◽  
Glutamyl Transpeptidase

When 4-aminophthalhydrazide is freshly diazotized and used as the coupling agent in demonstrating aminopeptidase activity or γ-glutamyl transpeptidase activity, with substrates that yield 4-methoxy-2-naphthylamine or even 2-naphthylamine, azo dyes are produced which are readily osmiophilic. Although the distribution of the reddish azo dyes is similar to that noted with earlier methods in light microscopy, the distribution of the deposits in electron microscopic preparations is distinctly membraneous in location in contrast to the droplet distribution of previous methods. We attribute membraneous localization of enzymatic activity with the new methods presented here to the lipophobic properties of the azo dyes provided by their 4-aminophthalhydrazide (4-APH) moiety. The new agent 4-aminophthalhydrazide was designed with this in mind.

Use of Uranium-Labeled Antibodies for Electron Microscopic Localization of Various Antigens in Mammalian Cells

1967 ◽  
Vol 25 ◽  
pp. 136-137
Author(s):  
J. P. Petrali ◽  
E. J. Donati ◽  
L. A. Sternberger
Keyword(s):  
Endoplasmic Reticulum ◽  
Hela Cells ◽  
Mammalian Cells ◽  
Specific Antiserum ◽  
Electron Microscopic ◽  
E Coli ◽  
Cytoplasmic Membranes ◽  
Viral Progeny ◽  
Ultrathin Sections ◽  
Electron Microscopic Localization

Specific contrast is conferred to subcellular antigen by applying purified antibodies, exhaustively labeled with uranium under immunospecific protection, to ultrathin sections. Use of Seligman’s principle of bridging osmium to metal via thiocarbohydrazide (TCH) intensifies specific contrast. Ultrathin sections of osmium-fixed materials were stained on the grid by application of 1) thiosemicarbazide (TSC), 2) unlabeled specific antiserum, 3) uranium-labeled anti-antibody and 4) TCH followed by reosmication. Antigens to be localized consisted of vaccinia antigen in infected HeLa cells, lysozyme in monocytes of patients with monocytic or monomyelocytic leukemia, and fibrinogen in the platelets of these leukemic patients. Control sections were stained with non-specific antiserum (E. coli).In the vaccinia-HeLa system, antigen was localized from 1 to 3 hours following infection, and was confined to degrading virus, the inner walls of numerous organelles, and other structures in cytoplasmic foci. Surrounding architecture and cellular mitochondria were unstained. 8 to 14 hours after infection, antigen was localized on the outer walls of the viral progeny, on cytoplasmic membranes, and free in the cytoplasm. Staining of endoplasmic reticulum was intense and focal early, and weak and diffuse late in infection.

The electron microscopic localization of nephrotoxic antibodies in isolated glomeruli

Pathology ◽  
1976 ◽  
Vol 8 (1) ◽  
pp. 73-80 ◽  
Author(s):  
I.P. McCausland ◽  
R.N. Seelye ◽  
J.B. Gavin ◽  
P.B. Herdson
Keyword(s):  
Electron Microscopic ◽  
Isolated Glomeruli ◽  
Microscopic Localization ◽  
Electron Microscopic Localization
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