Effect of experimental diabetes on ornithine decarboxylase activity of rat tissues

Measurements have been made of the activity of ornithine decarboxylase of liver, heart, kidney and brain in alloxan-diabetic and control rats. In all these tissues this enzyme had decreased markedly at four weeks after induction of diabetes. These results are discussed in relation to the hormonal control and cyclic nucleotide regulation of ornithine decarboxylase.     

Effect of 6-Aminonicotinamide on the activity of hexokinase and lactate dehydrogenase isoenzymes in regions of the rat brain

Changes in the activity of hexokinase and lactate dehydrogenase isoenzymes in the three brain regions and heart were studied in the 6-Aminonicotinamide-treated rats. Drug administration decreased the particulate hexokinase and lactate dehydrogenase activity, but increased the soluble hexokinase     

All hexokinase isoenzymes coexist in rat hepatocytes.

The cellular distribution of hexokinase isoenzymes, N-acetylglucosamine Kinase and pyruvate kinases in rat liver was studied. Hepatocytes and non-parenchymal cells with high viability and almost no cross-contamination were obtained by perfusion in situ of the liver with collagenase, with the use of an enriched cell-culture medium in all steps of cell isolation. Separation of hexokinase isoenzymes was done by DEAE-cellulose chromatography, and enzyme activities were measured by a specific radioassay. Cytosol from isolated hepatocytes contained high-affinity hexokinases A, B and C, in addition to hexokinase D. The last-mentioned represented about 95% of total glucose-phosphorylating activity. Only hexokinase A was found associated t the particulate fraction. Isolated non-parenchymal cells contained only hexokinases A, B and C. N-Acetylglucosamine kinase was measured with a specific radioassay and was found as a single enzyme form in both hepatocytes and non-parenchymal cells, with higher activities in the former. Pyruvate kinase isoenzyme L was present only in the hepatocytes and isoenzyme K only in the non-parenchymal liver cells, confirming that they are good cellular markers.     

Influence of streptozotocin-induced diabetes on hexokinase activity of rat salivary glands

The influence of diabetes on the enzyme hexokinase (HK) was examined in the salivary glands of rats. Diabetes was induced by an intraperitoneal injection of streptozotocin (60 mg/Kg body weight) in overnight fasted rats (180-200 g). The animals were killed 48 hours and 30 days after the induction of diabetes and the submandibular and parotid salivary glands extracted for use. Hyperglycemia was evaluated by determining the blood sugar. The area occupied by each intralobular component, acini, ducts, total parenchyma and stroma was measured, and no differences were observed compared with control. In the soluble fraction of the submandibular gland, no difference in the specific activity of HK was observed, between the diabetic and control animals, however, the activity per gland and per g of tissue showed lower values than control. The specific activity of the bound form was reduced in the diabetic gland. The results obtained for the parotid gland were different from the submandibular. The specific activity of both the soluble and bound forms were increased in the diabetic animals. The DEAE-cellulose column chromatography of the soluble and bound forms of the enzyme from both glands showed a first peak appearing during the washing of the column and two other peaks were eluted by the gradient. Thus, three isoenzymes in the submandibular and parotid salivary glands for the control and diabetic rats have been found.     

Effect of streptozotocin-induced diabetes on the turnover of hexokinase II in the rat

Skeletal muscle hexokinase II activity and turnover rates were measured in the normal and streptozotocin-induced diabetic rat. Enzyme activity decreases in the diabetic animal relative to the normal rat; however, the specific activity of hexokinase II is essentially the same for the two conditions. No alteration is observed in the relative rate of hexokinase II synthesis in the normal or diabetic rats, but there is a 3-fold increase in the rate of hexokinase II degradation in the latter group of animals. These results suggest that the primary cause of the well-established decrease in hexokinase II activity in skeletal muscle of the diabetic is an increase in the rate of enzyme degradation.     

Histochemical and immunohistochemical localization of hexokinase isoenzymes in rat kidney

Summary Histochemical and immunohistochemical staining techniques have been used to investigate the localization of hexokinase isoenzymes within rat kidney tissue. Hexokinase type I was shown to be the major isoenzyme present. It was located mainly in the thin and thick limbs of loops of Henle, in distal tubules and in the transitional or dark cells in the initial portions of collecting ducts. The smooth muscle cells of arteries and arterioles, peripheral nerves and the transitional epithelial cells lining the renal pyramid also contained large amounts of the isoenzyme while smaller quantities were present in glomeruli and in collecting tubules near the papillary tip. The distribution pattern obtained in tubular epithelia agrees well with that demonstrated in earlier microdissection studies. It is also consistent with the suggestion that glycolysis provides the majority of the energy fuelling the sodium transport mechanisms which form such an essential feature of the countercurrent urine concentration system present within the renal medulla.     

A study of hexokinase isoenzymes in rat sarcoma M-1

Using a wide spectrum of criteria, the isozyme composition of hexokinase from sarcoma M-1 reinoculated to rat m. gastrocnemius was studied. The structural, physicochemical and functional properties of the homogeneous enzyme which is represented in sarcoma M-1 by one molecular form, were investigated. Some properties of the enzyme (amino acid composition, resistance to mild proteolysis, Mr, pH-dependence of enzyme activity, electrophoretic mobility, kinetic behaviour) indicate that sarcoma M-1 hexokinase is a specific form of the enzyme which differs markedly from other known isozymes of mammalian hexokinase. The observed peculiarities of sarcoma M-1 hexokinase are discussed in terms of present-day concepts on the structure of isozymic spectra of enzymes in neoplastic tissues.     

Histochemical and immunohistochemical localization of hexokinase isoenzymes in normal rat liver

Summary Histochemical and immunohistochemical procedures have been used to examine the localization of three of the four hexokinase isoenzymes present in the liver of fed female Wistar rats. Distinctive distribution patterns were found for hexokinase type I and glucokinase but hexokinase type II was not detectable. Hexokinase type I was identified in sinusoidal cells and in bile duct epithelia, nerves and arteries in the portal triad. Glucokinase, the major isoenzyme, was confined to parenchymal cells where it was present in much higher amounts in perivenous compared with periportal hepatocytes. Staining within these two zones was not homogeneous and each had a mosaic appearance caused by the presence of a few hepatocytes containing little or no glucokinase amongst the majority of darkly stained cells in perivenous areas and a few darkly stained cells amongst the majority of unstained cells in periportal areas. Hence, hepatocytesin situ are a strikingly heterogeneous population of cells. Their metabolic status cannot be controlled simply by the differential supply of oxygen, substrates and hormones to different regions of the liver acini as proposed in the metabolic zonation model. Phenotypic differences may exist between cells within a given metabolic zone which influence their ability to respond to different environmental conditions.     

Changes in the subcellular distribution of brain and heart hexokinase isoenzymes during alloxan diabetes

Changes in the subcellular distribution of hexokinase activity from three brain regions and heart were studied during alloxan induced diabetes. There was an overall decrease in the particulate hexokinase with an increase in the soluble form, after different time intervals of the onset of diabetes. Administration of insulin to the diabetic rats showed a partial counteraction of the enzyme changes. A possible regulation of brain hexokinase by metabolite changes is proposed     

Changes in hexokinase isoenzymes in regions of rat brain during thyroid deficiency

In this work, activities of hexokinase isoenzymes Type I and Type II were measured in the soluble and particulate fractions from the brain regions (cerebral hemispheres (cerebrum), cerebellum and brain stem) of the thyroidectomized adult rats as well as of the thyroidectomized rats administered with triiodothyronine. Thyroidectomy generally decreased the hexokinase activity associated with particulate and soluble fractions. Hexokinase Type II isoenzyme was more affected than the Type I isoenzyme. Administration of triiodothyronine to the hypothyroid rats abolished the effect of thyroidectomy. Adult brain enzymes have been generally considered not be affected by thyroid hormones. The data obtained in this work are suggestive of an effect of thyroid hormones on hexokinase in the adult brain. Since the effects of thyroidectomy on the energy metabolism of the heart tissue are well known, the heart tissue was also studied for comparison.     

An inverse relation between mitochondrial hexokinase content and phosphoglucomutase activity of rat tissues

Summary The hexokinase: fumarase ratios of mitochondria isolated from ten tissues of the rat were determined, and compared with the tissue content of phosphoglucomutase and phosphorylase, taken as representatives of enzymes concerned with glycogen metabolism. A generally inverse relationship was found between the mitochondrial hexokinase: fumarase ratio and phosphoglucomutase levels. The cytochrome: fumarase ratios were relatively invariant in these same mitochondria. The results are interpreted as indicating a specialization of mitochondria, with increased amounts of hexokinase being associated with the mitochondria in tissues exhibiting less dependence on glycogen metabolism, as judged from phosphoglucomutase levels.     

The localization of hexokinase isoenzymes in red and white skeletal muscles of the rat

Summary Maximum assayable hexokinase activities vary with the proportion of red, fast-twitch, oxidative-glycolytic and intermediate, slow-twitch, oxidative fibres in different rat skeletal muscles. The major isoenzymic form, type II hexokinase, is present throughout the intermyofibrillar sarcoplasm in all fibres but a proportion of the total activity appears to be weakly associated with mitochondria. Variations in the histochemical staining intensity between fibre types correlate with their mitochondrial content and seem to be due mainly to differences in mitochondrially-associated hexokinase activity. Changes in the strength of this association may be important in controlling increases in glucose metabolism in response to prolonged increased muscular activity while regulation of the equilibrium between free and loosely-bound forms may be an important control feature in all skeletal muscle. Type I hexokinase is a minor isoenzymic component of skeletal muscle and occurs mainly in blood vessels and nerves in the perimysia and endomysia. The majority of this isoenzyme is tightly bound to mitochondria and is not detectable in homogenates prepared in the absence of Triton X-100.