scholarly journals A Histochemical Method for the Demonstration of Diphosphopyridine Nucleotide Diaphorase

1958 ◽  
Vol 4 (1) ◽  
pp. 29-38 ◽  
Author(s):  
Marvin M. Nachlas ◽  
Donald G. Walker ◽  
Arnold M. Seligman
Keyword(s):  
Gastric Mucosa ◽  
Rat Kidney ◽  
Succinic Dehydrogenase ◽  
Lactic Dehydrogenase ◽  
Histochemical Demonstration ◽  
Cytochemical Localization ◽  
Dehydrogenase System ◽  
Special Distribution ◽  
Almost All ◽  
Mucous Lining

The present investigation concerning the histochemical demonstration of DPN diaphorase follows the development of a new reagent, Nitro-BT, which has already been used successfully for the cytochemical localization of the succinic dehydrogenase system. The most consistently favorable results were obtained with the lactate-lactic dehydrogenase system buffered at pH 7.4. Using sections of rat kidney and stomach, it was found that the intensity of stain was optimal after 15 minutes incubation at 37°C., conducted aerobically. By appropriate variations in the substrate mixture it was possible to selectively demonstrate the histochemical distribution of certain DPN-linked dehydrogenases in addition to DPN diaphorase. This was made possible by the special distribution of some of these dehydrogenases which distinguished them from one another. Of the dehydrogenases studied the distribution pattern of ß-hydroxybutyric dehydrogenase was the most singular. In the gastric mucosa ß-hydroxybutyric dehydrogenase was restricted to the cells of the mucous lining epithelium and the gland necks; and in the kidney the enzyme was limited to the cells of the proximal convoluted tubule and thick limbs of Henle's loop. In contrast, lactic dehydrogenase like DPN diaphorase was demonstrable in almost all cytologic elements of both the stomach and the kidney.

ENZYME INHIBITION BY DERIVATIVES OF PHENOTHIAZINE: III. CATALASE, CYTOCHROME OXIDASE, AND DEHYDROGENASES

1942 ◽  
Vol 20b (12) ◽  
pp. 284-290 ◽  
Author(s):  
H. Bruce Collier ◽  
Della E. Allen
Keyword(s):  
Cytochrome Oxidase ◽  
Succinic Dehydrogenase ◽  
Lactic Dehydrogenase ◽  
Acid Oxidase ◽  
Amino Acid Oxidase ◽  
Leuco Form ◽  
Dehydrogenase System ◽  
Living Organisms ◽  
Relationship Of ◽  
Derivatives Of

The inhibition of liver catalase and of cytochrome oxidase by leucophenothiazone has been confirmed by manometric methods. Phenothiazine sulphoxide is a powerful catalase inhibitor, its activity being much greater at pH 5.3 than at neutrality.The succinoxidase activity of beef heart is inhibited by phenothiazone and by thionol. Phenothiazone in its oxidized form acts upon the succinic dehydrogenase, while the leuco form inhibits the cytochrome oxidase.Phenothiazone is reduced by the yeast lactic dehydrogenase system, and the enzyme activity is markedly decreased. Urease is also partially inhibited, whereas the effect of phenothiazone upon d-amino-acid oxidase is almost negligible.The possible relationship of these findings to the action of phenothiazine upon living organisms is discussed.

A Dye Phosphate for the Histo- and Cytochemical Demonstration of Alkaline Phosphatase, with some Observations on the Differential Behaviour of Nuclear and Extranuclear Enzymes

1949 ◽  
Vol s3-90 (9) ◽  
pp. 57-66
Author(s):  
A. LOVELESS ◽  
J. F. DANIELLI
Keyword(s):  
Alkaline Phosphatase ◽  
Rat Kidney ◽  
Histochemical Demonstration ◽  
Optimal Conditions ◽  
Enzymic Hydrolysis ◽  
Cytochemical Localization ◽  
Cytochemical Demonstration ◽  
End Products ◽  
Differential Behaviour ◽  
Hydrolysis Of

1. A new method is described for the cyto- and histochemical demonstration of alkaline phosphatase (monoesterase), a synthetic substrate p-nitrobenzene-azo-4α-naphthol-phosphate being employed as the sodium salt. Two methods for preparing a solution of this substance are described, and optimal conditions for enzymic hydrolysis of the ester worked out by the use of kidney sections. 2. Experiments are also described in which the substrate has been used to demonstrate phosphatase activity in calcifying tissues of dogfish and rat. 3. The results of the experiments with rat kidney indicate that traces of end-products are necessary before the enzyme can attack this substrate. 4. When results obtained with the new substrate are compared with those obtained with β-glycerophosphate, it is found that some sites display more activity towards one substrate than to the other. 5. The cytochemical localization of phosphatase with the new substrate is not as precise as with β-glycerophosphate.

scholarly journals HISTOCHEMICAL DEMONSTRATION OF THE SITES OF ACTIVITY OF DEHYDROGENASE SYSTEMS WITH THE ELECTRON MICROSCOPE

1957 ◽  
Vol 3 (4) ◽  
pp. 577-588 ◽  
Author(s):  
Russell J. Barrnett ◽  
George E. Palade
Keyword(s):  
Succinic Dehydrogenase ◽  
Potassium Cyanide ◽  
Histochemical Demonstration ◽  
Potassium Tellurite ◽  
Thin Sections ◽  
Fine Particulate ◽  
Close Relationship ◽  
Dehydrogenase System ◽  
Endogenous Activity ◽  
Enzymatic Nature

In the present study a histochemical method demonstrating the activity of dehydrogenase systems was developed for electron microscopy, utilizing potassium tellurite as the hydrogen or electron acceptor. This reagent was used intravitally (intravenously, intraperitoneally, or intraluminally in hollow organs) or supravitally on small blocks of tissue for the demonstration of endogenous dehydrogenase activity. Blocks of tissue which had been frozen and thawed or which had been washed in 0.44 M sucrose to prevent endogenous activity, were used to demonstrate the activity of the succinic dehydrogenase system. In the latter case, the incubating medium contained tellurite, succinate, phosphate buffer, sucrose, and activators. The incubation was as performed either aerobically (with or without the addition of potassium cyanide) or anaerobically. The specificity and the enzymatic nature of the reactions were ascertained by appropriate control experiments. Reduced tellurite, the end product of this histochemical reaction, could be visualized in thin sections of osmium tetroxide-fixed, methacrylate-embedded tissues as crystals or fine particulate deposits of high density, localized on, or in close relationship to mitochondrial membranes. The results of these experiments are demonstrated, utilizing heart muscle (rat) as the source of the enzyme systems.

Succinic and Malic Oxidase in Gastric Hydrochloric Acid Production

1956 ◽  
Vol 187 (3) ◽  
pp. 427-431 ◽  
Author(s):  
Joseph J. Vitale ◽  
Oscar M. Jankelson ◽  
Patricia Connors ◽  
D. Mark Hegsted ◽  
Norman Zamcheck
Keyword(s):  
Gastric Mucosa ◽  
Guinea Pig ◽  
Succinic Dehydrogenase ◽  
Titratable Acidity ◽  
Spectrophotometric Analysis ◽  
Parietal Cells ◽  
Human Gastric Mucosa ◽  
City Hospital ◽  
Malic Dehydrogenase ◽  
Oxidase System

Effect of histamine on the activity of succinic oxidase and malic dehydrogenase was studied in guinea pig and human gastric mucosa. Human tissue was obtained through the surgical services of the Boston City Hospital. Control value for the succinic oxidase system of the proximal half of the guinea pig stomach was approximately 480 ( Qo2 (N) (µl O2/mg nitrogen/hr.)). After histamine, this value rose to 550 in 30 minutes with a simultaneous rise in titratable acidity of the stomach contents. Animals fasted for 72 hours had a Qo2 (N) of approximately 500 and after histamine a Qo2 (N) of 700 was observed. Spectrophotometric analysis of succinic dehydrogenase and cytochrome oxidase activities, two of the major components of the succinic oxidase system, revealed that both components are increased following histamine administration. Malic dehydrogenase, however, was not affected by histamine treatment. Succinic dehydrogenase was demonstrated by histochemical localization and was concentrated below the superficial mucous layer where parietal cells were abundant. Succinic oxidase activity of human gastric mucosa was demonstrable only in those specimens containing abundant parietal cells. This study confirms the view that HCl production by parietal cells is associated with aerobic metabolism and is perhaps under enzymatic control. The study suggests that the succinic oxidase system may be involved in the production or secretion of HCl.

Formation of a Labile Pigment in Rabbit Ova During Histochemical Demonstration of Succinic Dehydrogenase

Science ◽  
1953 ◽  
Vol 117 (3043) ◽  
pp. 449-451 ◽  
Author(s):  
A. G. Foraker ◽  
S. W. Denham ◽  
D. D. Mitchell
Keyword(s):  
Succinic Dehydrogenase ◽  
Histochemical Demonstration

The Histochemical Demonstration of Succinic Dehydrogenase

Science ◽  
1951 ◽  
Vol 113 (2934) ◽  
pp. 317-320
Author(s):  
Arnold M. Seligman ◽  
Alexander M. Rutenburg
Keyword(s):  
Succinic Dehydrogenase ◽  
Histochemical Demonstration

scholarly journals The ultrastructural cytochemistry of lactic dehydrogenase, succinic dehydrogenase, dihydro-nicotinamide adenine dinucleotide diaphorase and cytochrome oxidase activities in hair cell mitochondria of the guinea pig cochlea.

1975 ◽  
Vol 23 (3) ◽  
pp. 216-234 ◽  
Author(s):  
G J Spector
Keyword(s):  
Hair Cell ◽  
Cytochrome Oxidase ◽  
Nicotinamide Adenine Dinucleotide ◽  
Succinic Dehydrogenase ◽  
Lactic Dehydrogenase ◽  
Nicotinamide Adenine ◽  
Fixation Method ◽  
Inner Mitochondrial Membrane ◽  
Tetrazolium Chloride ◽  
Outer Compartment

The use of cinnamyl nitroblue tetrazolium chloride (DS-NBT) in dehydrogenase experiments (lactic dehydrogenase, succinic dehydrogenase, nicotinamide adenine dinucleotide diaphorase) and 3,3'-diaminobenzidine tetrahydrochloride (DAB) in cytochrome oxidase experiments indicated that mitochondrial oxidoreduction reactions from nicotinamide adenine dinucleotide to cytochrome oxidase are located on the inner mitochondrial membrane in the outer compartment and the intracristate spaces. These reactions behave according to the chemiosmotic hypothesis. The cochlear hair cell mitochondria are cytochemically indistinguishable from free liver mitochondria. The heterogeneous mitochondrial staining pattern is related to the osmolarity of the incubation media, solubility of the enzymes and pH of the medium, but not to the fixation method.

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