scholarly journals CYTOPHOTOMETRIC QUANTITATION OF GLUCOSE 6-PHOSPHATASE ACTIVITY IN RAT LIVER

1969 ◽  
Vol 17 (2) ◽  
pp. 107-109 ◽  
Author(s):  
D. E. ABRAHAMSON ◽  
J. L. RIGATUSO ◽  
A. LAZAROW
Keyword(s):  
Phosphatase Activity ◽  
Rat Liver ◽  
Optical Density ◽  
Incubation Time ◽  
Specific Inhibitor ◽  
Histochemical Demonstration ◽  
Precipitation Method ◽  
Normal Tissues ◽  
Fresh Frozen ◽  
Normal Rat

The Wachstein-Meisel lead sulfide precipitation method was used for the histochemical demonstration of glucose 6-phosphatase activity in fresh-frozen normal rat liver sections. Transmittances, microspectrophotometrically determined over cytoplasmic areas of stained sections, were used to calculate the optical density of the stain. In spite of the fact that no absorption maximum was found, the spectrum of glucose 6-phosphatase stain showed proportional increases with increased incubation time. The extinction at a fixed wavelength was shown to be a linear function of incubation time. Studies in which normal tissues were incubated in buffer-substrate and then removed and reincubated for an additional period of time in buffer-substrate containing a specific inhibitor of glucose 6-phosphatase indicate that the increase in optical density of the stain was a result of glucose 6-phosphatase activity.

scholarly journals Freeze substituted tissue in 5'-nucleotidase histochemistry. Comparative histochemical and biochemical investigations.

1979 ◽  
Vol 27 (12) ◽  
pp. 1582-1587 ◽  
Author(s):  
K Klaushofer ◽  
H von Mayersbach
Keyword(s):  
Enzyme Activity ◽  
Sensitivity And Specificity ◽  
Rat Liver ◽  
Freeze Substitution ◽  
High Sensitivity ◽  
Histochemical Demonstration ◽  
Frozen Sections ◽  
Enzyme Localization ◽  
Fresh Frozen ◽  
Preparation Techniques

The suitability of freeze-substitution in n-butanol and paraffin embedding of tissues for the histochemical demonstration of 5'-nucleotidase was investigated and compared with commonly used preparation techniques, such as fresh frozen sections and cryostate sections of cold formalin and glutaraldehyde-fixed rat liver. The influences of each step of the preparation techniques on the enzyme activity were controlled by a quantitative radiochemical assay. Freeze substitution was revealed to be superior to the commonly used preparation techniques with respect to: 1) high sensitivity and specificity of the histochemical 5'-nucleotidase reaction (this is based on the fact that incubation media with very low lead concentrations (0,6 mM/1) can be used); 2) excellent morphological appearance of the tissues showing cytological details of enzyme localization; 3) unlimited storage of the tissue materials and ease of sectioning.

scholarly journals A NEW HISTOCHEMICAL METHOD FOR ACID PHOSPHATASE BY THE USE OF 5-IODOINDOXYL PHOSPHATE

1966 ◽  
Vol 14 (2) ◽  
pp. 171-176 ◽  
Author(s):  
G. W. EVANS ◽  
CECILIA L. WHINNEY ◽  
K. C. TSOU
Keyword(s):  
Acid Phosphatase ◽  
Phosphatase Activity ◽  
Acid Phosphatase Activity ◽  
Azo Dye ◽  
Redox System ◽  
Histochemical Demonstration ◽  
Crystal Formation ◽  
Frozen Sections ◽  
Fresh Frozen ◽  
Ph Range

5-Iodoindoxyl phosphate has been found to be a useful indigogenic substrate in the histochemical demonstration of acid phosphatase activity. Its superiority to other indoxyl phosphates is apparently due to a rapid oxidation of 5-iodoindoxyl to 5,5'-diiodoindigo in the acid pH range. A redox system of ferri-ferrocyanide enhances the oxidation and improves the localization. This method can be applied to calcium-formol-fixed tissues or to fresh frozen sections, although fixed tissues yield better results. The method is not recommended for the demonstration of enzyme activity in lipid-rich tissues because of the complexing property of lipids with 5,5'-diiodoindigo that results in crystal formation. The distribution of acid phosphatase activity with this method is generally similar to that obtained using azo dye methods.

scholarly journals LOCALIZED AREAS OF HIGH ALKALINE PHOSPHATASE ACTIVITY IN THE TERMINAL ARTERIAL TREE

1962 ◽  
Vol 15 (1) ◽  
pp. 73-84 ◽  
Author(s):  
Flaviu C. A. Romanul ◽  
Roger G. Bannister
Keyword(s):  
Alkaline Phosphatase ◽  
Enzymatic Activity ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Histochemical Demonstration ◽  
Capillary Endothelium ◽  
Parent Vessel ◽  
Arterial Tree ◽  
Fresh Frozen ◽  
Cryostat Sections

Fresh frozen skeletal muscles of rats, rabbits, and humans were sectioned in a cryostat. Sections 12 to 32 micra thick were incubated in a substrate solution for the histochemical demonstration of non-specific alkaline phosphatase activity. A modified azo dye coupling technique was used at pH 9.5. Localized areas of high enzymatic activity were found in specific and well defined areas along the terminal arterial tree, in addition to the activity which has been previously described in capillary endothelium. Arterial branches with luminal diameters of 25 micra or less showed staining of their endothelium starting abruptly at their origin from the parent vessel and fading distally. Smaller arterial branches showed the same localization of enzymatic activity and stained more intensely. Other organs of rats surveyed showed arterial branches with the same pattern of staining. Identical results were obtained using the Gomori technique for alkaline phosphatase. Extensive saline perfusion of the vascular tree did not affect the observed localization of enzymatic activity. The enzymatic activity described may be part of the mechanism regulating the blood flow.

scholarly journals STUDIES ON THE FUNCTION OF INTRACELLULAR RIBONUCLEASES

1966 ◽  
Vol 29 (3) ◽  
pp. 395-403 ◽  
Author(s):  
Takeshi Utsunomiya ◽  
Jay S. Roth
Keyword(s):  
Natural Products ◽  
Sodium Chloride ◽  
Rat Liver ◽  
Messenger Rna ◽  
Rnase Activity ◽  
Optimum Ph ◽  
Normal Rat ◽  
Pancreatic Rnase ◽  
The Stability ◽  
Normal Constituent

The RNase activity and properties of ribosome and polysome preparations from normal rat liver and some hepatomas have been examined. Polysome and ribosome preparations from the Novikoff, McCoy MDAB, and Dunning hepatomas had considerably higher specific RNase activity than corresponding preparations from normal rat liver, Novikoff ascites, or Morris 5123 hepatomas. The optimum pH of the RNase was approximately 8.5 for all samples tested, and the samples showed no evidence of latent RNase activity when treated with 3 M sodium chloride, EDTA, urea, or p-chloromercuribenzenesulfonic acid. The RNase activity appeared to be associated principally with breakdown products and/or subunits smaller than 80S. In the presence of Mg++ ions, subunits could reaggregate to form monomer ribosomes indistinguishable from the natural products, but some of the reassociated ribosomes could contain RNase activity which had been bound to the smaller particles. Similar results were obtained with spermine. In the hepatomas, evidence was obtained for the preexistence of considerable amounts of the smaller, RNase-containing subunits in the cell. When a small amount of crystalline bovine pancreatic RNase was added to partly dissociated ribosomes, the RNase was found only in association with the smaller subunits, and little or no enzyme was taken up by ribosomes or polysomes. The results have led to the conclusion that RNase is not a normal constituent of the ribosome or polysome, but that RNase may become associated with these particulates if dissociation and reassociation take place. Some implications of these findings for the stability of messenger RNA and for the mechanism of its breakdown are discussed.

Intracellular re-localisation by photochemical internalisation enhances the cytotoxic effect of gelonin — Quantitative studies in normal rat liver

2010 ◽  
Vol 142 (3) ◽  
pp. 347-353 ◽  
Author(s):  
Josephine Woodhams ◽  
Pei-Jen Lou ◽  
Pål K. Selbo ◽  
Alexander Mosse ◽  
Dahmane Oukrif ◽  
...  
Keyword(s):  
Rat Liver ◽  
Cytotoxic Effect ◽  
Quantitative Studies ◽  
Normal Rat

scholarly journals Retinyl palmitate hydrolase activity in normal rat liver.

1981 ◽  
Vol 256 (9) ◽  
pp. 4498-4503
Author(s):  
J.H. Prystowsky ◽  
J.E. Smith ◽  
D.S. Goodman
Keyword(s):  
Rat Liver ◽  
Retinyl Palmitate ◽  
Hydrolase Activity ◽  
Normal Rat

Hepatoma, host liver, and normal rat liver phospholipids as affected by diet

Lipids ◽  
1975 ◽  
Vol 10 (12) ◽  
pp. 736-745 ◽  
Author(s):  
Randall Wood
Keyword(s):  
Rat Liver ◽  
Host Liver ◽  
Normal Rat
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