scholarly journals HISTOCHEMICAL DEMONSTRATION OF MITOCHONDRIAL ADENOSINE TRIPHOSPHATASE ACTIVITY IN TISSUE SECTIONS

1962 ◽  
Vol 10 (1) ◽  
pp. 65-74 ◽  
Author(s):  
MAX WACHSTEIN ◽  
MAIRE BRADSHAW ◽  
JOSÉ M. ORTIZ
Keyword(s):  
Adenosine Triphosphatase ◽  
Histochemical Demonstration ◽  
The Other ◽  
Mitochondrial Activity ◽  
Bile Canaliculi ◽  
Adenosine Triphosphatase Activity ◽  
Tissue Sections ◽  
Dog Kidney ◽  
Cryostat Sections ◽  
Neutral Formalin

A comparative study was made of the distribution of mitochondrial adenosine triphosphatase activity in several organs of rat and rabbit using both the calcium and lead techniques. Certain modifications in these techniques assured both improved intracellular morphology and considerable preservation of enzyme activity. Of particular importance was the low temperature (–2 to –3°C) of the neutral calcium-formol solution, and, following short fixation (lead method), a subsequent wash in neutral buffer. Mitochondrial activity was similar with both techniques, but the lead method proved to be less costly, less time-consuming, and above all, far less capricious than the calcium technique. In good preparations, the appearance of mitochondria is as clearly defined as in sections stained with non-enzymatic, conventional techniques. Long exposure of cryostat sections to formalin or preparation of sections from tissue blocks fixed in neutral formalin leads to the complete abolition of mitochondrial activity; on the other hand, it accentuates enzyme staining of other structures, such as, for instance, bile canaliculi in the liver and the secretory capillaries in the pancreas and salivary glands. It also visualizes the infolding membranes in certain tubules of the rat and dog kidney. It is assumed that formalin-fixation aids in the enzymato-morphologic distinction of these two different intracellular structures.

The histochemical demonstration of myofibrillar adenosine triphosphatase activity in fish muscle

1974 ◽  
Vol 52 (7) ◽  
pp. 871-877 ◽  
Author(s):  
I. A. Johnston ◽  
S. Patterson ◽  
P. Ward ◽  
G. Goldspink
Keyword(s):  
Atpase Activity ◽  
Adenosine Triphosphatase ◽  
Fish Muscle ◽  
Histochemical Demonstration ◽  
Differential Staining ◽  
Fiber Types ◽  
Myofibrillar Atpase ◽  
Adenosine Triphosphatase Activity ◽  
Acid Ph ◽  
Myofibrillar Atpase Activity

A technique for the demonstration of myofibrillar adenosine triphosphatase activity (ATPase) used for mammalian muscle has been modified to suit fish muscle. The mammalian method involves selectively inhibiting fiber types by preincubation at either alkaline (pH 10.4) or acid (pH 4.3) pH before incubation for myofibrillar (ATPase) activity. Fish muscle fibers were found to be generally inactivated under these conditions. Preincubation at an acid pH was found to be unsuitable for fish muscle because of the indiscriminate inactivation of the fibers. The effects of preincubating at pH 10.4 and incubating tissue sections for different time periods and at different pH's and temperatures have been investigated. A differential staining of fiber types correlated with biochemical data on myofibrillar ATPase for red and white muscles was obtained by preincubating sections for short periods (1–2 min) at pH 10.4. Under these conditions the intermediately positioned pink fibers were found to stain similarly to the white fibers of high myofibrillar ATPase activity. An investigation has been made of the qualitative distribution of fiber types in the myotomal muscle of live teleost species: coalfish (Gadus virens), grey mullet (Mugil cephalus), crucian carp (Carassius carassius), black mollie (Mollienesia sp), and glassfish (Chanda ranga). The pink fibers were found to be abundant in all the species examined with the exception of the glassfish.

scholarly journals ADENOSINE TRIPHOSPHATASE: A NEUROHISTOCHEMICAL STUDY

1962 ◽  
Vol 10 (6) ◽  
pp. 731-740 ◽  
Author(s):  
D. NAIDOO
Keyword(s):  
Atpase Activity ◽  
Adenosine Triphosphatase ◽  
Wistar Rat ◽  
The Other ◽  
Nuclear Staining ◽  
Adenosine Triphosphatase Activity ◽  
Bivalent Cations ◽  
Vascular Elements ◽  
The Brain ◽  
Cerebral Vascular

The location of adenosine triphosphatase in the brain has been studied in rapidly frozen-dried cerebral tissues of the Wistar rat. It is found that adenosine triphosphatase is an almost exclusively nuclear enzyme. Two tissue fractions of the cerebrum were separated, so that one sample was made up of vascular elements, and the other of neural elements. The two fractions were then studied for their adenosine triphosphatase activity, and compared with the histochemical findings. The two tissue fractions were found not to differ in the absence of bivalent cations. When Ca++ were added to the cerebral vascular suspension, ATPase activity was increased approximately 15 times, and only 3 times in the presence of Mg++. Conversely, the addition of Mg++ increased the ATPase activity of the neural fraction 200%; whereas, Ca++ was responsible for a 60% increase. This fact was detectable microscopically when Ca++ was found to intensify vascular nuclear staining, and Mg++ to increase the neuronal and glial nuclear staining. The results, histochemical and biochemical, are mutually confirmatory.

scholarly journals Temperature-dependence of activation and inhibition of rat-brain adenosine triphosphatase activated by sodium and potassium ions

1966 ◽  
Vol 100 (3) ◽  
pp. 762-767 ◽  
Author(s):  
N Gruener ◽  
Y Avi-Dor
Keyword(s):  
Rat Brain ◽  
Adenosine Triphosphatase ◽  
Arrhenius Plot ◽  
Enzyme System ◽  
The Other ◽  
Potassium Ions ◽  
Adenosine Triphosphatase Activity ◽  
Brain Microsomes ◽  
High Concentrations ◽  
Sodium And Potassium

1. The adenosine-triphosphatase activity of rat-brain microsomes was measured between 0 degrees and 37 degrees . The stimulatory effect of Na(+) plus K(+) on the Mg(2+)-dependent adenosine-triphosphatase activity decreased sharply with decreasing temperature and became negligible at 0 degrees . An Arrhenius plot drawn from the experimental data showed two discontinuities: one at about 6 degrees and the other at about 20 degrees . 2. The increment in activity induced by Na(+) plus K(+) was more sensitive to oligomycin at lower than at higher temperatures, but the opposite was observed for ouabain. The action of oligomycin showed a biphasic character, since below a certain concentration it caused slight activation of Na(+)-plus-K(+)-activated adenosine triphosphatase. 3. Where oligomycin increased the activity of the enzyme, it also enhanced the accumulation of an acid-precipitable phosphorylated compound formed through the transfer of the gamma-phosphate group of [(32)P]ATP to the enzyme system. Stimulatory concentrations of oligomycin did not interfere with K(+)-mediated dephosphorylation of the intermediate, though high concentrations of oligomycin counteracted the effect of K(+). 4. The temperature profile of K(+)-stimulated microsomal phosphatase qualitatively resembled that of microsomal adenosine triphosphatase.

scholarly journals Microspectrophotometric quantitation of the diaminobenzidine reaction for histochemical demonstration of cytochrome oxidase activity.

1978 ◽  
Vol 26 (3) ◽  
pp. 157-162 ◽  
Author(s):  
A C Frasch ◽  
M E Itoiz ◽  
R L Cabrini
Keyword(s):  
Cytochrome Oxidase ◽  
Incubation Medium ◽  
Histochemical Demonstration ◽  
Tongue Muscle ◽  
Tissue Sections ◽  
Direct Light ◽  
Fixed Concentration ◽  
Set Up ◽  
Cryostat Sections

Polyacrylamide models in which an extract of cattle heart mitochondria was incorporated, as well as cryostat sections of tongue muscle and epithelium, were used to set up the conditions under which the histochemical reaction for the demonstration of cytochrome oxidase can be quantitated. Using diaminobenzidine in a concentration of 5.5 mM, cytochrome C in a fixed concentration of 76 micron and keeping the incubation medium away from direct light action, enzyme activity can be evaluated by means of direct microphotometry on tissue sections. Each biologic model requires previous individual determination of the measurement limits. These limits can be readily established by using a small chamber for the incubation medium, which can be placed in the microphotometer, allowing the reaction rate to be following using a single section.

scholarly journals THE HISTOCHEMICAL DEMONSTRATION OF ADENOSINE TRIPHOSPHATASE ACTIVITY IN MYOCARDIUM

1964 ◽  
Vol 12 (10) ◽  
pp. 740-743 ◽  
Author(s):  
N. R. NILES ◽  
J. CHAYEN ◽  
G. J. CUNNINGHAM ◽  
LUCILLE BITENSKY
Keyword(s):  
Enzymatic Activity ◽  
Adenosine Triphosphate ◽  
Adenosine Triphosphatase ◽  
Inhibitory Effect ◽  
Histochemical Demonstration ◽  
Human Myocardium ◽  
Phosphate Esters ◽  
Precise Localization ◽  
Adenosine Triphosphatase Activity

Adenosine triphosphatase has been demonstrated histochemically in rat and human myocardium. To obtain its precise localization in discrete bands, apparently corresponding to the concentration of myosin, it was necessary to modify the existing technique to obtain better preservation of unfixed tissue and maximal enzymatic activity. Thus it was necessary to increase the concentration of calcium and to effect the reaction at pH 9.4 after treatment with 2:4-dinitrophenol. The specificity of the reaction was shown by these factors, by testing with phosphate esters other than adenosine triphosphate, and by the inhibitory effect of magnesium.

scholarly journals A NEW DIAZO REAGENT FOR SPECIFIC STAINING OF CONJUGATED BILIRUBIN IN TISSUE SECTIONS

1968 ◽  
Vol 16 (6) ◽  
pp. 419-427 ◽  
Author(s):  
V. J. DESMET ◽  
A.-M. BULLENS ◽  
J. DE GROOTE ◽  
K. P. M. HEIRWEGH
Keyword(s):  
Total Bilirubin ◽  
Diazonium Salt ◽  
Histochemical Demonstration ◽  
Specific Method ◽  
Bile Pigments ◽  
Specific Staining ◽  
Tissue Sections ◽  
A New Technique ◽  
Cryostat Sections ◽  
Conjugated Bilirubin

A new technique is presented for the specific histochemical demonstration of conjugated bilirubin in tissue sections, using the diazonium salt of ethylanthranilate. The specificity of this method was proved by using test materials and cholestatic tissue sections. There was no diffusion artifact under the prescribed working conditions. The method is applicable to fresh cryostat sections and frozen sections of cold, formol-calcium-fixed tissues. The method could not be satisfactorily applied for the staining of total bilirubin. The method using 2,4-dichloraniline remains the best available technique for the histochemical demonstration of total bilirubin, while the technique with ethylanthranilate proves to be the most specific method available for the demonstration of conjugated bile pigments.

scholarly journals Histochemical demonstration of neurotoxic esterase.

1989 ◽  
Vol 37 (5) ◽  
pp. 589-596 ◽  
Author(s):  
G B Koelle ◽  
N S Thampi ◽  
M S Han ◽  
E J Olajos
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Enzymatic Activity ◽  
Histochemical Demonstration ◽  
The Other ◽  
Delayed Neurotoxicity ◽  
The Central Nervous System ◽  
Cryostat Sections ◽  
Insoluble Precipitate ◽  
Neurotoxic Esterase

We developed a histochemical method for localizing neurotoxic esterase (NTE), defined as the phenylvalerate (PV)-hydrolyzing esterase that is resistant to 40 microM paraoxon (A) but inactivated by paraoxon plus 50 microM mipafox (B). NTE is considered to be the target enzyme in the production of organophosphorus ester-induced delayed neurotoxicity (OPIDN). Cryostat sections were incubated in a medium containing alpha-naphthyl valerate and 6-benzamido-4-methoxy-m-toluidine diazonium chloride (fast violet B) after treatment with the above-mentioned inhibitors, leading to formation of an aqueous insoluble precipitate at sites of enzymatic activity. NTE activity was estimated as staining detectable in A but not in B. In the central nervous system (CNS) of chicken, NTE appeared to be present primarily in the somata of most neurons, but at sites indistinguishable from those of the other inhibitor-resistant and -sensitive alpha-naphthyl valerate-hydrolyzing esterases. It could not be distinguished in the CNS of cat, probably because it constitutes less than 3% of the total PV-hydrolyzing activity in the CNS of that species.

scholarly journals Isolation and properties of brain α-actinin

1978 ◽  
Vol 175 (1) ◽  
pp. 63-72 ◽  
Author(s):  
W Schook ◽  
C Ores ◽  
S Puszkin
Keyword(s):  
Sodium Dodecyl Sulphate ◽  
Gel Electrophoresis ◽  
Synaptic Vesicles ◽  
Adenosine Triphosphatase ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Bovine Brain ◽  
The Other ◽  
Muscle Actin ◽  
Adenosine Triphosphatase Activity

alpha-Actinin isolated from bovine brain migrated on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis like muscle alpha-actinin with an apparent mol.wt. of 100000 and cross-reacted with antibodies to muscle alpha-actinin. Brain alpha-actinin modulated actin-myosin Mg2+-activated adenosine triphosphatase activity and, when bound by polystyrene particles, was found to bind muscle actin and tropomyosin from solution. Brain alpha-actinin, in conjunction with the other components of the contractile and relaxing complex, may play a role in the release of neurotransmitters from synaptic vesicles.

scholarly journals Genetic complementation between two mutant unc alleles (unc A401 and unc D409) affecting the F1 portion of the magnesium ion-stimulated adenosine triphosphatase of Escherichia coli K12

1978 ◽  
Vol 170 (3) ◽  
pp. 593-598 ◽  
Author(s):  
G B Cox ◽  
J A Downie ◽  
F Gibson ◽  
J Radik
Keyword(s):  
Escherichia Coli ◽  
Electron Transport ◽  
Fluorescence Quenching ◽  
Oxidative Phosphorylation ◽  
Mutant Allele ◽  
Adenosine Triphosphatase ◽  
The Other ◽  
Genetic Complementation ◽  
Magnesium Ion ◽  
Adenosine Triphosphatase Activity

A new mutant strain of Escherichia coli in which phosphorylation is uncoupled from electron transport was isolated. A genetic-complementation analysis, using partial diploid strains, showed that the new mutant allele, uncD409, is in a gene distinct from the other previously identified genes uncA, uncB and uncC. A strain carrying the uncd409 allele has no Mg2+ ion-stimulated adenosine triphosphatase activity and is therefore phenotypically similar to strains carrying the uncA401 mutant allele. Complementation between the uncA401 and the uncD409 alleles occurred, as indicated by growth of partial diploid strains on succinate and their growth yields on limiting concentrations of glucose. Complementation was confirmed by using membranes prepared from the above partial diploids. Such membranes were found to have Mg2+-stimulated adenosine triphosphatase activity, ATP-dependent transhydrogenase activity ADP-induced atebrin-fluorescence quenching and low but significant amounts of oxidative phosphorylation.

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