scholarly journals MICROPHOTOMETRIC DETERMINATION OF ENZYME ACTIVITY IN SINGLE CELLS IN CRYOSTAT SECTIONS I. APPLICATION OF THE GEL FILM TECHNIQUE TO MICROPHOTOMETRY AND STUDIES ON THE INTRALOBULAR DISTRIBUTION OF SUCCINATE DEHYDROGENASE AND LACTATE DEHYDROGENASE ACTIVITIES IN RAT LIVER

1972 ◽  
Vol 20 (8) ◽  
pp. 567-576 ◽  
Author(s):  
J. NOLTE ◽  
D. PETTE
Keyword(s):  
Enzyme Activity ◽  
Lactate Dehydrogenase ◽  
Succinate Dehydrogenase ◽  
Dehydrogenase Activity ◽  
Reaction Rate ◽  
Single Cells ◽  
Succinate Dehydrogenase Activity ◽  
Activity Determination ◽  
Gel Film ◽  
Cryostat Sections

A gel film technique was adopted for kinetic enzyme activity determination in single cells using cryostat sections by means of a microscope photometer. The main principle of the method is a comparative activity determination, based on bipositional recording of initial reaction kinetics in two preselected measuring fields in the same tissue section. Systematic model experiments were performed to prove the fulfillment of the following conditions: ( a) validity of Beer's law; ( b) optimal concentrations of substrates, cosubstrates and dye within the reaction film, including evidence that the reaction rate is not limited by insufficient diffusion of these compounds; ( c) proportionality between local enzyme concentration or thickness of tissue sections and the recorded reaction rate; and ( d) specificity of the method as demonstrated by studying reaction rates in the absence of the substrate as well as in the presence of substrate and a specific inhibitor. The validity of the method was also examined by comparing the levels of succinate dehydrogenase activity in various rat tissues, as measured microphotometrically, with the enzyme levels as determined in homogenates. Microphotometric assays for nicotinamide adenine dinucleotide phosphate isocitrate dehydrogenase, lactate dehydrogenase and succinate dehydrogenase are described. Using these techniques it was found that: the periportal and central areas of the hepatic lobule in rat contain the same level of lactate dehydrogenase activity; succinate dehydrogenase activity is 1.6 times higher in the periportal area as compared to the centrilobular area. In experimental thyrotoxicosis, the ratio of enzyme activities in the periportal and central areas was 1.1.

scholarly journals MICROPHOTOMETRIC DETERMINATION OF ENZYME ACTIVITY IN SINGLE CELLS IN CRYOSTAT SECTIONS II. SUCCINATE DEHYDROGENASE, LACTATE DEHYDROGENASE AND TRIOSEPHOSPHATE DEHYDROGENASE ACTIVITIES IN RED, INTERMEDIATE AND WHITE FIBERS OF SOLEUS AND RECTUS FEMORIS MUSCLES OF RAT

1972 ◽  
Vol 20 (8) ◽  
pp. 577-582 ◽  
Author(s):  
J. NOLTE ◽  
D. PETTE
Keyword(s):  
Lactate Dehydrogenase ◽  
Succinate Dehydrogenase ◽  
Dehydrogenase Activity ◽  
Soleus Muscle ◽  
Cross Sections ◽  
Single Cells ◽  
Rectus Femoris ◽  
Succinate Dehydrogenase Activity ◽  
Parallel Increase ◽  
Musculus Soleus

Comparative microphotometric determinations of succinate dehydrogenase, triosephosphate dehydrogenase and lactate dehydrogenase activities in single white, red and intermediate fibers in cross-sections of rat rectus femoris muscle and soleus muscle were performed. White fibers have the highest triosephosphate and lactate dehydrogenase activities and the lowest succinate dehydrogenase activity. Red fibers are highest in succinate dehydrogenase activity and relatively low in triosephosphate and lactate dehydrogenase activities. The intermediate fibers have medium succinate dehydrogenase activity and relatively low triosephosphate dehydrogenase activity. In the soleus muscle, the triosephosphate dehydrogenase activity of the red fibers exceeds that of the intermediate fibers. A parallel increase in succinate dehydrogenase is observed in the red fibers of musculus soleus and musculus rectus femoris after thyroid hormone administration. There is some evidence, however, that the red fibers of the two muscles investigated vary with regard to their absolute succinate dehydrogenase activities and also the triosephosphate dehydrogenase to succinate dehydrogenase activity ratio.

Some enzymes of carbohydrate metabolism in Mesocestoides corti and Heterakis spumosa

1991 ◽  
Vol 65 (3) ◽  
pp. 187-192 ◽  
Author(s):  
P. Dubinský ◽  
B. Rušcinová ◽  
S. L. Hetmanski ◽  
C. Arme ◽  
L. Turčeková ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Lactate Dehydrogenase ◽  
Succinate Dehydrogenase ◽  
Carbohydrate Metabolism ◽  
Dehydrogenase Activity ◽  
Malate Dehydrogenase ◽  
Adult Females ◽  
Enzymes Of Carbohydrate Metabolism

ABSTRACTThe activities of selected enzymes of carbohydrate metabolism were measured in tetrathyridia of Mesocestoides corti and in adult females and males of Heterakis spumosa. When the species were compared, only lactate dehydrogenase and phosphoenolphyruvate carboxykinase activities were considerably higher in M. corti. Activities of other enzymes were higher in H. spumosa, with malate dehydrogenase activity being considerable so. In H. spumosa, enzyme activity was higher, and succinate dehydrogenase markedly so in males, when compared with females. Tetrathyridia aged 170 and 210 days show relatively stable malate and lactate dehydrogenase activities, and mice of ICR and BALB/c strains are suitable for the maintenance of tetrathyridia.

Effects of chronic hypoxia and dietary restriction on myocardial enzyme activities

1976 ◽  
Vol 231 (4) ◽  
pp. 1308-1313 ◽  
Author(s):  
SE Barrie ◽  
P Harris
Keyword(s):  
Lactate Dehydrogenase ◽  
Succinate Dehydrogenase ◽  
Dehydrogenase Activity ◽  
Dietary Restriction ◽  
Enzyme Activities ◽  
Chronic Hypoxia ◽  
Phosphoglycerate Kinase ◽  
Barometric Pressure ◽  
Succinate Dehydrogenase Activity ◽  
Left And Right

Enzyme activities were measured in homogenates of left and right ventricles of guinea pigs after 14 and 28 days' exposure to 400 mmHg barometric pressure. All animals developed anorexia and right ventricular hypertrophy. Two control groups of animals were used, one free fed and the other restricted to the amount of food chosen by the hypobaric group. The factorial design of the experiment allowed some distinction between the effects of anorexia, hypertrophy, and hypoxia. Dietary restriction was associated with a decrease in glycogen phosphorylase, hexokinase, and succinate dehydrogenase activity and an increase in the M-subunits of lactate dehydrogenase. Myocardial hypertrophy was associated with an increase in the activity of the enzymes of the glycolytic pathway down as far as phosphoglycerate kinase and an increase in the M-subunits of lactate dehydrogenase. Chronic hypoxia seemed specifically to be associated with an increase in the H-subunits of lactate dehydrogenase and possibly a slight transient increase in succinate dehydrogenase activity. Mixing studies indicated that changes in enzyme activities were likely to be due to changes in enzyme concentrations.

Interactive effects of emphysema and malnutrition on diaphragm structure and function

1994 ◽  
Vol 77 (2) ◽  
pp. 947-955 ◽  
Author(s):  
M. I. Lewis ◽  
S. A. Monn ◽  
W. Z. Zhan ◽  
G. C. Sieck
Keyword(s):  
Succinate Dehydrogenase ◽  
Dehydrogenase Activity ◽  
Cross Sectional Area ◽  
Interactive Effects ◽  
Type I ◽  
Specific Force ◽  
Type Ii ◽  
Sectional Area ◽  
Succinate Dehydrogenase Activity ◽  
Cross Sectional

Interactive effects of emphysema (EMP) and prolonged nutritional deprivation (ND) on contractile, morphometric, and metabolic properties of hamster diaphragm muscle (DIA) were examined. Six months after induction of EMP (intratracheal elastase), saline-treated controls (CTL) and EMP hamsters of similar body weights were subjected to ND over 6 wk. Isometric contractile and fatigue properties of costal DIA were determined in vitro. DIA fibers were histochemically classified as type I or II, and fiber succinate dehydrogenase activity and cross-sectional area were determined using quantitative microscopic procedures. From histochemical sections, the number of capillaries per fiber (C/F) and per fiber cross-sectional area (C/A) were determined. ND resulted in progressive loss of body weight (ND-CTL, 23.8%; ND-EMP, 28.4%; P = NS). ND did not affect reduction in optimal length (Lo) of DIA fibers in EMP compared with CTL and ND-CTL hamsters. Maximum specific force (i.e., force/unit area) was reduced by approximately 25% in EMP animals compared with CTL. ND did not improve or exacerbate the reduction in specific force with EMP. ND attenuated improved fatigue resistance of DIA in EMP animals. No differences in fiber type proportions were noted among experimental groups. Significant atrophy of type I and II DIA fibers was noted after ND. Atrophy was proportionately greater in type II fibers of ND-EMP when referenced to EMP animals. Thus adaptive hypertrophy of type II DIA fibers in EMP animals was abolished. Fiber succinate dehydrogenase activity was significantly increased in type I and II fibers in EMP DIA. ND did not affect this metabolic adaptation of DIA fibers to persistent loads imposed by EMP.(ABSTRACT TRUNCATED AT 250 WORDS)

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