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D Sasse

Publications and source records attributed to D Sasse.

At least 37 records · Page 2Linked to original sources

The circadian rhythm of intra-acinar profiles of alcohol dehydrogenase activity in rat liver: a microquantitative study.

Using microquantitative measurements of alcohol dehydrogenase activity in microdissected samples of liver tissue along the sinusoidal length, the intra-acinar distribution profiles were studied in seven groups of female rats at different times during 24 h with a light phase from 6:30 h to 18:30 h. The mean values of alcohol dehydrogenase activity showed a circadian rhythm with a minimum at 13.30 h and a maximum at 17.30 h (p less than 0.001). However, the intra-acinar gradients remained almost unchanged, indicating that increase and decrease in enzyme activity takes place simultaneously in all parts of the liver acinus. This observation, together with data from the literature, suggests that the circadian rhythm of alcohol dehydrogenase activity reflects variations in different liver cell constituents, rather than enzyme protein synthesis or proteolysis.

Alcohol Dehydrogenase↗

Nutritional and gonadal effects on the intra-acinar profiles of low-Km and high-Km aldehyde dehydrogenase activity in rat liver.

Total and low-Km aldehyde dehydrogenase (ALDH) activity was measured in 50-150 ng microdissected liver tissue samples of the entire sinusoidal length. High-Km ALDH activity was calculated by subtracting the low-Km ALDH values from the total ALDH activity. Enzyme activity was measured by a microchemical assay, using the oil-well technique with luminometric determination of NADH. The intra-acinar profiles of high-Km and low-Km ALDH activity could be demonstrated graphically for both male and female rats after 84 h of starvation, and after starvation and refeeding for 6 nights. In addition, the ALDH distribution patterns of juvenile, castrated, and castrated and testosterone-treated rats were determined. It could be demonstrated that starvation, and starvation followed by refeeding, lead to changes in enzyme activity which parallel the loss and regain of liver- and body-weight. The nutritional factors do not essentially alter the normal intra-acinar profiles. In juvenile rats, ALDH is lower by 30% in comparison with the controls, but sex-differences in the distribution profiles are not yet present. Castration has no effect on the amount of enzyme activity but the sex specific distribution profiles are less marked. The main effect of testosterone treatment is an elevation of low-Km ALDH in the perivenous zone. The characteristics of the intra-acinar profiles of high-Km and low-Km ALDH activity are discussed with respect to hepatic acetaldehyde oxidation and alcoholic liver damage.

Aldehyde Dehydrogenase↗

Microquantitative determination of intra-acinar distribution profiles of low-Km and high-Km aldehyde dehydrogenase activity in rat liver.

Microquantitative measurements of total and of low-Km aldehyde dehydrogenase (ALDH) activity with millimolar and micromolar concentrations of acetaldehyde and propionaldehyde were carried out on the livers of male and female rats. Lyophilized cryostat sections of liver parenchyma were microdissected along the entire sinusoidal length from the terminal afferent vessels to the terminal efferent venule. ALDH activity was measured in a microbiochemical assay using the oil-well technique with luminometric determination of NADH. On the basis of single measurements, mean values of total, low-Km and high-Km ALDH activity could be calculated and the specific distribution patterns graphically demonstrated. The two substrates acetaldehyde and propionaldehyde yielded similar values of ALDH activity, the intraacinar distribution profiles of which showed characteristic sex differences. In the liver of the male rat high-Km ALDH activity has two flat peaks in the periportal and the perivenous area, while low-Km ALDH activity is almost evenly distributed throughout the acinus. In the livers of female rats, both high-Km and low-Km ALDH activity shows a continuous gradient which decreases from the periportal to the perivenous zone (pp/pv = 1.4:1). It was therefore possible to demonstrate that the maxima of alcohol dehydrogenase activity and of low-Km ALDH activity are localized in opposite parts of the liver acinus of the female rat. This heterotopy should have consequences with respect to hepatotoxicity after alcohol ingestion.

Aldehyde Dehydrogenase↗

The effects of starving and refeeding on the intra-acinar distribution pattern of alcohol-dehydrogenase activity in rat liver.

Microquantitative determinations of ADH activity were carried out on the livers of male and female rats. The animals were either starved for 84 h, or starved and then refed with a carbohydrate-rich diet for 6 nights. When the enzyme activity is expressed in mumoles/min/g dry weight, fasting does not appear to alter liver ADH activity, while in starved and subsequently refed rats it is diminished by 20%. Microquantitative measurements of ADH activity in 50-150 ng lyophilized tissue samples, microdissected the whole way along the sinusoidal length, made the computer-aided plotting of intra-acinar distribution patterns possible. The results showed that, under the feeding conditions selected, only minor changes in the ADH activity profiles occur in the liver acinus. These are within the range of the standard deviations of the normal mean values. From these results it can be deduced that fasting and refeeding do not lead to specific inhibition or induction of liver ADH activity. - The decrease of ADH activity of total liver (mumol/min) per total body weight in starved rats is obviously the result of a loss of protein which affects the liver cells of all acinar zones almost equally.

Alcohol Dehydrogenase↗

The intra-acinar distribution patterns of alcohol-dehydrogenase activity in the liver of juvenile, castrated and testosterone-treated rats.

Rat liver alcohol dehydrogenase shows characteristic sex-differences with respect to activity and heterotopy. For the recognition of gonadal influences on the intra-acinar distribution patterns luminometric determinations of ADH activity were carried out on 50-150 ng lyophilized liver tissue samples which had been microdissected along the sinusoidal length. Juvenile rats of both sexes showed equally high alcohol dehydrogenase activity, which surpassed the adult values by a factor of 2 in males and 1.3 in females. The distribution pattern was rather flat, with a weak maximum at the beginning of the last third of the sinusoid. Castration of adult male and female rats resulted in an increase of alcohol dehydrogenase activity to around the prepubertal values. The intra-acinar profiles showed a gradual increase in activity from low periportal values to a peak near the perivenous zone. Only the hepatocytes directly adjacent to the efferent venule showed an even lower activity. Administration of testosterone to castrated animals had no effect on the ADH activity in males and resulted in only a slight decrease of enzyme activity in females. The intra-acinar distribution patterns showed an intermediary peak at the end of the second third of the sinusoidal length in males and a gradual increase of activity, beginning periportally, in the direction of the perivenous zone in females. The present findings on total activity of ADH and its distribution patterns in the liver are considered to be the result of complex hormonal alterations rather than a specific effect of testosterone.

Age Factors↗

The relation of rat liver wet weight to dry weight.

To assess a reliable relation between the dry and wet weight of rat livers, the water content of liver samples was determined by freeze drying. The ratio between wet and dry weight of the livers turned out as 3.33 +/- 0.3 for male and 3.28 +/- 0.24 for female rats. Thus for calculations a value of 3.3 can be used irrespective of the sex of rats.

Animals↗

Histochemical studies on metabolic zonation of the liver in the trout (Salmo gairdneri).

The livers of 26 adult male and female trout were studied histochemically. G6Pase activity was always found to be heterotopically distributed with a constant maximum in the periportal area. In many cases the glycogen content and the activity of phosphorylase predominated in the periportal zone as well. Maximum activity of glucose-6-phosphate-dehydrogenase and malic enzyme, however, could be demonstrated preferentially in the perivenous area. Lactate dehydrogenase, succinate dehydrogenase, alcohol dehydrogenase, acid phosphatase and beta-glucuronidase were found equally in all liver cells. 3-Hydroxybutyrate dehydrogenase was absent. Thus, the principles of metabolic zonation have been established in trout liver, the architecture of which differs essentially from that of mammals. The course of the terminal afferent and efferent vessels is the decisive factor for the heterotopic localization of functional units rather than the tubular or plate-forming arrangement of the hepatocytes.

Acid Phosphatase↗

Microquantitative determination of the distribution patterns of alcohol dehydrogenase activity in the liver of rat, guinea-pig and horse.

Microquantitative measurements of ADH-activity were carried out on the livers of male and female rats, guinea-pigs and horses (two geldings and a mare). Lyophilized cryostat sections of liver parenchyma were microdissected the whole way along the sinusoidal length from the terminal afferent vessels to the terminal efferent venule. ADH activity in samples of about 50-150 ng was measured in a microbiochemical assay using the oil-well technique without enzymatic cycling, by direct luminometric determination of NADH. On the basis of the single measurements, mean values of total hepatic ADH activity could be calculated and the specific distribution patterns graphically demonstrated. Total activity of ADH in the liver of the female rat is 1.6 times higher than in the male; the male distribution pattern exhibits a relative maximum in the intermediary zone of the acinus while the activity in the liver of female rats increases towards a perivenous maximum. Mean values for total ADH activity in the livers of male and female guinea-pigs are almost equal and there is, moreover, no clear intra-acinar gradient. Mare and castrated male horses show high hepatic ADH activity which is evenly distributed in the liver acinus.

Alcohol Dehydrogenase↗

Antagonistic reaction of the periportal and perivenous zone in the rat liver after castration and estrogen treatment. Histochemical and biochemical studies on G6PDH and malic enzyme activity.

G6PDH and ME activity was determined biochemically in homogenates and demonstrated histochemically in cryostat sections of rat liver. Control animals were sham-operated, the male and female rats of the experimental groups were castrated. After castration groups of rats were treated with daily doses of 3 or 6 micrograms/estradiol benzoate for 21 days. The results show that in the controls there is a sex-dependent distribution pattern of the two enzymes; in males the rather low activity is mainly located in the periportal area, in females the higher activity is demonstrable in the perivenous area. After castration G6PDH activity (and to a lesser extent ME activity) increases, mainly in the periportal zone. Estrogen treatment results in the high activity of both enzymes, which are exclusively located in the perivenous zone. In the periportal zone no G6PDH or ME activity is demonstrable histochemically. This zone-typical effect of estrogen is interpreted in terms of the concept of Metabolic Zonation, according to which it is supposed that the NADPH generating enzymes in the perivenous area have a lipogenic function whereas the periportal activity contributes to bile acid production.

Animals↗

[Postnatal adaptation of gluconeogenic and liponeogenic areas in the liver parenchyma].

The activity of G6Pase points to the gluco(neo)genic function of hepatocytes, whereas the activity of G6PDH and malic enzyme, both yielding NADPH, are involved in liponeogenesis. Histochemical studies of the distribution patterns and quantitative measurements concerning the postnatal weeks of rats show that after the 6th week sex differences occur. In contrast to males in female rat liver a lipogenic area emerges in the perivenous zone. This lipogenic zone is situated opposite to the gluco(neo)genic area which corresponds to the periportal zone.

Animals↗

The glucose/glucose-6-phosphate cycle in the periportal and perivenous zone of rat liver.

Periportal and perivenous hepatocytes contain different activities (V) of antagonistic key enzymes such as glucokinase and glucose-6-phosphatase. In order to get an insight into the metabolism of the periportal and perivenous area the flux rates (v) of the glucose/glucose-6-phosphate cycle were calculated on the basis of the Michaelis-Menten equation using the measured zonal concentrations of glucose and glucose 6-phosphate, the zonal activities of glucokinase and glucose-6-phosphatase previously reported and the half-saturating substrate concentrations (Km) of the two enzymes found in the literature. The concentrations of glucose were obtained as a first approximation by measuring the concentrations in portal (= periportal) and hepatovenous (= perivenous) blood; those of glucose 6-phosphate were calculated from the levels determined in microdissected periportal and perivenous liver tissue. The calculations showed (a) that the overall cycling rates agreed remarkably well with those reported for intact animals and (b) that during a normal feeding rhythm the periportal zone should catalyze net glucose output and the perivenous zone should mediate net glucose uptake, as proposed by the model of 'metabolic zonation'.

Animals↗

Postnatal differentiation of sex-specific distribution patterns of G6Pase, G6PDH and ME in the rat liver.

The activity of the liver enzymes G6Pase, G6PDH and ME was studied in rats of 2-9 weeks old by histochemical means. In addition, G6PDH and ME activity was quantitatively determined in homogenates. In the 2nd and 3rd week G6Pase is similarly distributed in both sexes: while in the periportal zone high activity is demonstrable, the perivenous zone shows only low activity. After this period a nearly homogeneous distribution pattern becomes evident in all animals. Sex difference occurs after the 6th week: in the livers of male rats the periportal "maximum" is sometimes combined with a second peak in the perivenous area, in females a steep gradient emerges with high activity in the periportal zone and a low one in the perivenous zone. In the first postnatal weeks G6PDH activity is very low in parenchymal cells, but very prominent in Kupffer cells. Around the 5th week there is an increase, predominantly in the perivenous zone of both sexes. While there is again a further decrease demonstrable in male rats, the G6PDH activity of female rats rises to high adult values. This increase seems to be restricted to the perivenous zone. ME can be demonstrated at first in leucocytes. In the course of the 3rd week there is an increase of activity in both sexes: ME is demonstrable in parenchymal cells of the perivenous area and in scattered hepatocytes of the periportal area. In male rats, the perivenous activity is diminished towards the end of the investigation period, in females, however, a high activity remains in the perivenous zone. The data show that in females the activity of NADP dependent enzymes is high in the perivenous zone, so it may be assumed that a lipogenic area is situated around the terminal efferent vessels. Because of the sex difference this area may be hormone-dependent. The lipogenic area is situated opposite to the gluco(neo)genic area which corresponds to the periportal zone.

Aging↗

Quantitative topography of organelles in the liver. A combined histochemical and morphometric analysis.

After seven days of feeding fructose the liver of Wistar rats showed enormous accumulations of glycogen, which completely altered the original pattern of distribution of organelles. A quantitative morphological method was used to analyze these changes. The cytoplasm was mapped into arbitrary "distance classes" corresponding to concentric rings beginning at the outer nuclear membrane. This allowed the density of organelles in a given zone to be estimated. In cells filled with glycogen as a result of the fructose feeding, the following rearrangements were found: in the intermediate zone of both cellular poles (i.e., bile canalicular pole and sinusoidal pole) the mitochondria disappeared, being replaced by glycogen. The endoplasmic reticulum was accumulated in the perinuclear zone of both cellular poles, as in control animals, but was reduced throughout the rest of cytoplasm. It showed a peripheral density maximum at the biliary canalicular pole, in contrast to the cells of control animals. These changes in the distribution of the organelles and cellular "compartments" correspond to histochemical findings and demonstrate an adaptive reaction in the liver parenchyma to fructose ingestion, the organelles arranging themselves in cytoplasmic regions which still show a metabolic activity.

Animals↗

Metabolic zonation in thioacetamide-induced liver cirrhosis.

After TAA administration to rats a central part may be distinguished histochemically from a marginal part in most of the cirrhotic nodules. The centre is characterized by a high glycogen content and by high activity of phosphorylase, G6Pase and SDH; the maxima of which are situated around the larger blood vessels. The vasculatory periphery, however, shows moderate G6PDH-activity. The marginal parts of the nodules are poor in glycogen and possess only weak G6Pase and phosphorylase activity, whereas high SDH- and G6PDH-activity can be demonstrated here. This distribution pattern leads to the conclusion that the larger blood vessels in the centre of the nodules are themselves the terminal afferent vessels. Thus the centre of the nodule corresponds to periportal zone 1, while G6PDH-activity marks the area corresponding to zone 3. The fact that the marginal parts of the nodules are marked by high SDH- but weak G6Pase-activity is interpreted as the result of a preferential arterial supply to this parenchymal part. The high G6PDH-activity of the marginal part is seen in context with the regeneration processes. In all animals single nodules could be found with a high glycogen content and extremely high G6PDH-activity. This loss of heterogeneity is interpreted as a first step in the direction of malignancy.

Animals↗