Effect of a high fat diet on rat adipocyte lipolysis: responses to epinephrine, forskolin, methylisobutylxanthine, dibutyryl cyclic AMP, insulin and nicotinic acid.

An earlier report from this laboratory showed that feeding rats a high fat diet decreased epinephrine-stimulated lipolysis in their adipose tissue. Experiments were designed to explore further the effects of such diets on adipocyte response to epinephrine and to several other lipolytic and antilipolytic agents. Rats were fed diets with 67% of energy consisting of glucose or lard for 5 to 7 d.

Effect of acute exercise on insulin generation of pyruvate dehydrogenase activator by rat liver and adipocyte plasma membranes.

Groups of young adult rats with body weights of 125-135 g (group A) or 300-400 g (group B) were subjected to one bout of prolonged exercise to exhaustion on a treadmill and were studied 2 h postexercise. Liver glycogen levels were markedly depleted in the exercised rats. Adipocytes from group A exercised rats showed a significantly greater increase in pyruvate dehydrogenase (PDH) activity in response to insulin than those from sedentary controls.

Effect of trifluoperazine on the action of insulin in rat adipocytes.

Trifluoperazine (TFP), a potent inhibitor of calmodulin action, at a concentration of 12 microM decreased the stimulating effects of insulin on 1) fat cell pyruvate dehydrogenase (PDH) activation, 2) generation/action of PDH activator by adipocyte plasma membranes, and 3) insulin-induced loss of insulin receptors, without altering spermine-induced activation of fat cell PDH or preventing insulin stimulation of glucose oxidation. In addition to these effects on insulin action, TFP abolished several biological actions of the insulin-generated PDH stimulator from liver particulate fractions. These actions include fat cell PDH activation and decrease in receptors.

Insulin-induced internalization and replacement of insulin receptors in adipocytes of rats adapted to fat feeding.

To elucidate the mechanisms of the previously observed decrease in adipocyte surface insulin binding in fat diet-induced insulin resistance (Ip et al., J. Lipid Res.

Studies on the effects of protease substrate analogues on some of the actions of insulin.

Added TAME (N alpha-p-tosyl-1-anginine methyl ester) or BAME (benzoyl-anginine methyl ester) inhibited insulin induced activation of glucose oxidation and fat cell PDH activation without affecting spermine action on PDH activation and glucose oxidation in fat cells. BAME inhibited insulin-induced generation of both PDH stimulator and PDH inhibitor from liver particulate fraction. In contrast, insulin-induced internalization of insulin receptors and negative cooperativity of insulin receptors were not affected by protease substrate inhibitors.

Studies on the possible involvement of prostaglandins in insulin generation of pyruvate dehydrogenase activator.

Insulin-exposed liver particulate fraction supernatants from control rats stimulated mitochondrial pyruvate dehydrogenase (PDH) activity by 26%, while the stimulation by similar preparations from indomethacin-injected rats (5 mg/kg twice daily, i.p., for 2 days) was 4%.

Mechanisms of the fasting-induced dissociation of insulin binding from its action in isolated rat hepatocytes.

Fasting leads to an increase in insulin binding to isolated rat hepatocytes from 12 to 17%. This increase was accounted for by changes in the affinity of insulin receptors without alteration in their number. In contrast, the responsiveness of hepatocytes to insulin was markedly diminished in fasted rats.

Effect of dexamethasone on adipose tissue and liver pyruvate dehydrogenase and its stimulation by insulin-generated chemical mediator.

Rats were treated with dexamethasone (50 micrograms/day, sc) for 4 days. Total pyruvate dehydrogenase (PDH) and insulin-stimulated PDHa activities were decreased in fat pads from dexamethasone-treated rats compared to control values. Coincubation experiments with adipocyte mitochondria, plasma membrane, and insulin demonstrated decreased stimulation of PDH in preparations from dexamethasone-treated rats.

Decreased insulin-generation of pyruvate dehydrogenase inhibitor in insulin resistant states.

Insulin resistance produced in rats by feeding a high fat diet or by dexamethasone administration (50 micrograms/day, sc for 4 days) resulted in 50-70% decrease in the generation of pyruvate dehydrogenase inhibitor by insulin exposed liver particulate fractions. The inhibition was dose dependent. Treatment of insulin mediator preparations with neuraminidase and B-D-galactosidase resulted in inactivation of the pyruvate dehydrogenase inhibitor.

The effects of streptozotocin diabetes on the activities of rat liver glycosyltransferases.

The effects of streptozotocin diabetes on the activities of rat liver glycosyltransferase enzymes have been investigated. Liver microsomal fractions were prepared from rats that had been injected with streptozotocin (65 mg/kg, i.v.

Effects of high fat and high carbohydrate diets on liver pyruvate dehydrogenase and its activation by a chemical mediator released from insulin-treated liver particulate fraction: effect of neuraminidase treatment on the chemical mediator activity.

Rats were fed a high fat diet or a high glucose diet for 5-7 days. Basal pyruvate dehydrogenase activity (both the active form and the total enzyme activity) was decreased in liver homogenates from fat diet-adapted rats as compared to those fed the glucose diet. Supernatants from insulin-exposed liver particulate fractions from fat-fed rats showed decreased stimulation of pyruvate dehydrogenase activity as compared to those from glucose-fed rats.

Effects of high carbohydrate and high fat diets on rat adipose tissue pyruvate dehydrogenase responses to concanavalin A and spermine.

Rats were fed a high lard diet or a high glucose diet for 5-7 days. Basal and Concanavalin A (Con A)-stimulated epididymal fat pad pyruvate dehydrogenase (PDH) activities were decreased in fat diet-adapted rats compared to those fed the glucose diet. When adipocyte plasma membranes and mitochondria were coincubated with and without Con A, it was found that the lectin stimulation of PDH activity was lower in preparations from fat-fed rats.

Effect of high fat and high carbohydrate diets on adipose tissue pyruvate dehydrogenase and its activation by a plasma membrane-enriched fraction and insulin.

Rats were fed a high lard diet or a high glucose diet for 5--7 days. Basal and insulin-stimulated epididymal fat pad pyruvate dehydrogenase (PDH) activities were decreased in fat diet-adapted rats compared to those fed the glucose diet. When adipocyte plasma membranes and mitochondria were incubated together with and without insulin, it was found that the insulin stimulation of PDH activity was lower in preparations from fat-fed rats on both an absolute and percentage basis.

Effects of high glucose and high lard diets on the activities of rat liver glycosyltransferases.

The activities of several enzymes involved in glycoprotein synthesis were measured in the livers of rats (L) fed diets with 67% of calories as lard and compared with those of rats (G) fed 67% glucose diets for 5-9 days. Glucosamine synthetase activity was not influenced by diet, but the activities of UDP-N-acetylglucosaminyl, galactosyl and sialyltransferases were significantly greater in the livers of the rats fed the glucose diet than in L rat livers. The content of UDP-N-acetylglucosamine was also higher in G livers than in the L group.

Dietary effects on the formation of dolichyl monophosphate mannose by microsomal preparations of rat adiopose tissue.

Microsomal preparations from rat adipose tissue catalyse the transfer of [(14)C]mannose from GDP-[(14)C]mannose to an endogenous acceptor forming a [(14)C]mannosyl lipid. The mannosyl lipid co-chromatographs with hen oviduct dolichyl monophosphate beta-mannose on three solvent systems. It is stable to mild alkaline hydrolysis, but strong alkaline treatment yields a compound that co-migrates with mannose 1-phosphate.

Lipolysis and cyclic AMP response to isoproterenol in diaphragms from control and dystrophic mice.

A comparison was made of the sensitivity of lipolysis (glycerol and free fatty acid release) and of cyclic AMP production to the action of isoproterenol in diaphragms from control and dystrophic Bar Harbor mice at 7 weeks of age. An increased lipolytic response was observed in diaphragms from dystrophic mice that was more apparent in the males, and was demonstrable when cyclic AMP was used instead of isoproterenol. The increased glycerol and free fatty acid release in response to isoproterenol and cyclic AMP cannot be explained by a higher triglyceride content of diaphragms from dystrophic mice, because it was found to be similar to that of controls when it was estimated by biochemical and light microscopic techniques.

Control of endogenous triglyceride breakdown in the mouse diaphragm.

The control of endogenous triglyceride breakdown was studied in vitro, in the incubated intact mouse diaphragm. Isoproterenol (2 microgram/ml) produced parallel increases in glycerol and free fatty acid release, and in tissue cyclic AMP levels, suggesting that cyclic AMP mediates the action of the catecholamine on triglyceride mobilization. In addition to cyclic AMP, calcium seems to be involved in the action of isoproterenol because preincubation of hemidiaphragms in the presence of the calcium ionophore A23187 decreased the lipolytic effect of the drug.

Studies on the utilization and mobilization of lipid in skeletal muscles from streptozotocin-diabetic and control rats.

Several aspects of lipid metabolism in the soleus and diaphragm muscles of streptozotocin-diabetic and control rats were investigated. The triglyceride content of both muscles was elevated in the diabetic state and the presence of increased intracellular lipid was confirmed by electron microscopy. In vitro glucose and palmitate oxidation studies showed that both types of muscle from the diabetic animals metabolized more fat than did the soleus and diaphragm from control rats.

Effects of high-glucose and high-fat diets on concanavalin A binding to rat liver plasma membranes and on the amount and pattern of their glycoprotein carbohydrates.

Purified liver plasma membranes were prepared from rats fed a high-fat, carbohydrate-free diet or a high-glucose, fat-free diet. Membranes from rats fed the high-fat diet bound significantly less 125I-labeled concanavalin A (Con A) than did those from rats fed the fat-free diets. The magnitude of the binding difference increased with increasing concentrations of Con A.

Lipid composition of liver plasma membranes isolated from rats fed a high glucose or a high fat diet.

Several plasma membrane associated functions have been shown to differ in rats fed a high lard, carbohydrate free diet (L rats) from those observed in preparations from rats fed a high glucose, fat free diet (G rats). To explore the possibility that differences in the lipid components of the plasma membranes might contribute to these functional changes, groups of rats were fed each diet for 5 days and the lipids of their plasma membranes were separated and analyzed. The major differences found were a greater cholesterol content in the plasma membranes from L rats compared to those of G rats (12.

The concentration of cyclic AMP and the activity of cyclic AMP dependent protein kinase and an inhibitor in the adipose tissue of rats fed lard or glucose diets.

Measurements of tissue cyclic AMP (cAMP) concentration, the activity of cAMP-dependent protein kinase and the level of the enzyme's thermostable, macromolecular inhibitor were made on preparations of rat epididymal fat pad from animals fed high fat or high carbohydrate diets. The cAMP concentration from rats adapted to a high lard diet for 14-15 days was 153 +/- 17.8 pmoles/mg protein as opposed to 76 +/- 6.

Studies on serum lipids, insulin, and glucagon and on muscle triglyceride in rats adapted to high-fat and high-carbohydrate diets.

A comparisonwas made of lipid circulation, storage, and mobilization in rats adapted to lard or glucose diets. In the morning, lard-fed rats had higher blood triglyceride (TG) and free fatty acid (FFA) levels. In the evening TG was higher, but FFA was significantly lower in the lard vs.