Biomedical subjects

A Sener

Publications and source records attributed to A Sener.

At least 199 records · Page 11Linked to original sources

PubMed1988

Branched chain alpha-ketoacid dehydrogenase and pyruvate dehydrogenase activity in isolated rat pancreatic islets.

Branched-chain alpha-ketoacid dehydrogenase and pyruvate dehydrogenase in isolated rat pancreatic islets were shown to be regulated by a phosphorylation/dephosphorylation mechanism. Broad-specificity phosphoprotein phosphatase treatment stimulated and ATP addition inhibited their activities. The kinases responsible for inactivating these complexes were shown to be sensitive to inhibition by known inhibitors, alpha-chloroisocaproate and dichloroacetate. Total activity (nmol/min/islet / 37 degrees C) of branched-chain alpha-ketoacid dehydrogenase and pyruvate dehydrogenase was 0.86 and 5.09, with a % active form (activity before phosphatase treatment divided by activity after phosphatase treatment X 100) of 36% and 94%, respectively. Incubation of intact isolated islets with alpha-chloroisocaproate affected neither insulin release nor flux through branched-chain alpha-ketoacid dehydrogenase.

3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)↗

PubMed1988

Metabolism of D-glucose anomers in rat lens.

In lens homogenates, hexokinase displayed a greater affinity for alpha-D-glucose but a higher maximal velocity with beta-D-glucose. However, in intact lenses incubated at 10 degrees C in the presence of 4-7 mM alpha- or beta-D-glucose, no anomeric difference could be found in either the intracellular content in D-glucose and D-glucose 6-phosphate or the utilization of D-[5-3H]glucose and oxidation of D-[1-14C]glucose. Even the sorbitol content of the lens was not significantly different after 180 min perifusion at 37 degrees C in the presence of either alpha- or beta-D-glucose. It is proposed that the lack of anomeric specificity in D-glucose metabolism is attributable, at least in part, to the low turnover rate of glucose 6-phosphate in the lens, allowing for the interconversion of glucose 6-phosphate anomers.

Animals↗

PubMed1988

Defective catabolism of D-glucose and L-glutamine in mouse pancreatic islets maintained in culture after streptozotocin exposure.

We recently described a preferential reduction of the secretory response to nutrient secretagogues (glucose; leucine plus glutamine) in islets maintained in culture after in vitro exposure to streptozotocin (SZ). The present study is an attempt to further clarify the biochemical mechanisms behind this defective insulin response. Mouse pancreatic islets were collagenase isolated and, after 4-5 days in culture, exposed during 30 min at 37 C to 1.8 mM SZ or vehicle alone (controls). The islets were subsequently cultured for 7 days in medium RPMI 1640 plus 10% calf serum, before the enzymatic and metabolic studies were performed. The activities of the glycolytic enzymes, hexokinase, glucokinase, and glyceraldehyde 3-phosphate dehydrogenase, were similar in the control and SZ-exposed islets. The relative amount of cytosolic and mitochondria-bound hexokinase was also unaffected by SZ. However, there was a 30-40% decrease in the activity of NAD+- and NADP+-dependent glutamate dehydrogenase and glutamate-aspartate transaminase in the SZ-treated islets. This coincided with a 40% decrease in L-[U-14C]glutamine oxidation in the SZ-treated islets. The D-glucose catabolism was further examined in the presence of D-[5-3H] and D-[6-14C] glucose. There was no difference between control and SZ islets in terms of glucose utilization at either 1.7 or 16.7 mM glucose. The oxidation of D-[6-14C]glucose was nevertheless decreased by more than 50% in SZ islets incubated at 16.7 mM (but not 1.7 mM) glucose. Altogether, these converging observations suggest a perturbation of distal regulatory processes, apparently at the mitochondrial level, in the D-glucose and L-glutamine catabolism of SZ-exposed islets. Whether this reflects a primary action of SZ on the islet mitochondria, or an inhibitory effect of SZ on the synthesis of mitochondrial enzymes, as a result of nuclear DNA damage, remains to be elucidated.

Animals↗

PubMed1988

Effects of L-leucine, its 2-keto acid metabolite and its non-metabolized analogue on rat tumoral islet cell function.

L-Leucine and 2-ketoisocaproate stimulated insulin release from perifused rat tumoral islet cells (RINm5F line). The secretory response coincided with an increase in the intracellular ATP/ADP ratio, a stimulation of 45Ca outflow from cells perifused in the presence of extracellular Ca2+, and an increase in 32P efflux from cells prelabelled with radioactive orthophosphate. In contrast to D-glucose, however, L-leucine or 2-ketoisocaproate failed to decrease 86Rb outflow, to inhibit 45Ca outflow from cells perifused in the absence of Ca2+ and to enhance the labelling of inositol-containing phospholipids in cells exposed to myo-[2-3H]inositol. These findings suggest that D-glucose, L-leucine and 2-ketoisocaproate exert dissimilar effects on the subcellular distribution of adenine nucleotides and/or 86Rb. The non-metabolized analogue of L-leucine, 2-aminobicyclo[2.2.1]heptane-2-carboxylic acid (BCH), also caused an initial stimulation of insulin release and 32P efflux, but this was soon followed by a severe and irreversible inhibition of insulin output, associated with a permanent enhancement of 86Rb outflow. The dual ionic and secretory response to BCH is interpreted in the light of its dual effect on the catabolism of endogenous amino and fatty acids, and raises the view that BCH could be used to interfere with the function of insulinoma cells.

Adenine Nucleotides↗

PubMed1988

Insulin production and glucose metabolism in isolated pancreatic islets of rats with NIDDM.

Rats with non-insulin-dependent diabetes mellitus (NIDDM) induced by neonatal injection of streptozocin are known to have a deficient insulin response to glucose. To evaluate to what extent this glucose insensitivity can be attributed to a perturbation of the islet glucose metabolism, we estimated the rates of glucose phosphorylation, glucose utilization, oxygen consumption, and glucose oxidation in islets isolated from normal and NIDDM rats and compared these values with rates of islet insulin biosynthesis and release in vitro. The data confirm that islets from rats with NIDDM display a deficient response to glucose of both insulin biosynthesis and release that is still present after an overnight culture of the islets at 5.5 mM glucose. Furthermore, they show that islets of these rats have 1) normal low- and high-Km glucose-phosphorylating activities and no major alteration of the glucose utilization rate, 2) decreased insulin release in response to glyceraldehyde, 3) decreased rates of basal respiration and glucose oxidation and a markedly reduced stimulation by glucose of both islet oxygen consumption and glucose oxidation, and 4) decreased glucose-stimulated net 45Ca uptake. We conclude that the relative unresponsiveness to glucose of islets from NIDDM rats is associated with, and perhaps due to, a deficient islet glucose metabolism. This defect is not due to gross alterations in the glycolytic pathway but probably reflects alteration in the islet mitochondria function.

Animals↗

PubMed1988

Hexose metabolism in pancreatic islets: the Pasteur effect.

In rat pancreatic islets, hypoxia severely decreased both the oxidation of D-[U-14C]glucose and the release of insulin evoked by D-glucose. The production of [14C]lactate was increased in the hypoxic islets, the relative magnitude of such an increment being greater at low (2.8 mM) than high (8.3 and 16.7 mM) D-glucose concentrations. Hypoxia increased the detritiation of D-[5-3H]glucose at low glucose concentration (2.8 mM), failed to affect 3H2O production at an intermediate glucose level (8.3 mM), and inhibited the utilization of D-[5-3H]glucose at a higher hexose concentration (16.7 mM). In tumoral islet cells (RINm5F line) exposed to 16.7 mM D-glucose, hypoxia decreased D-[U-14C]glucose oxidation to the same extent as in normal islet cells, but increased the production of [14C]lactate and 3H2O to a greater extent than in normal islets. These findings indicate that the Pasteur effect is operative in islet cells. The experimental data also suggest that, under normal conditions of oxygenation, high concentrations of D-glucose lead to both activation of phosphofructokinase and stimulation of mitochondrial oxidative events in normal, but not tumoral, islet cells.

Adenoma, Islet Cell↗

PubMed1988

The coupling of metabolic to secretory events in pancreatic islets: does hexose transport affect cationic fluxes?

Poorly metabolized hexoses, such as 3-O-methyl-D-glucose, 2-deoxy-D-glucose and D-galactose failed to reproduce the inhibition of 86Rb outflow, the early inhibition and secondary rise in 45Ca efflux and the stimulation of insulin release evoked by D-glucose in perifused rat islets. Insulin release induced by either D-glucose or 2-ketoisocaproate was also unaffected by 3-O-methyl-D-glucose. It is concluded that hexose transport in islet cells does not represent in itself a significant determinant of the cationic and secretory response to D-glucose.

3-O-Methylglucose↗

PubMed1987

Fructose metabolism via the pentose cycle in tumoral islet cells.

In tumoral islet cells (RINm5F line) the phosphorylation of D-fructose is catalyzed by hexokinase rather than fructokinase. Fructose 6-phosphate appears to be preferentially channelled into the pentose cycle, as suggested by a ratio of D-[1-14C]fructose/D-[U-14C]fructose oxidation close to 2.7, the failure to generate 14C-labelled lactate from D-[1-14C]fructose and a poor metabolic response to menadione. When the islet cells are exposed to both D-fructose and D-glucose, however, the metabolism of the former hexose is dramatically modified, fructose 6-phosphate being now formed at a lower rate and preferentially channelled into the glycolytic pathway. These findings illustrate the existence of regulatory steps in fructose catabolism located distally to its site of phosphorylation.

Adenoma, Islet Cell↗

PubMed1987

Influence of D-glucose upon the respiratory and secretory response of insulin-producing tumor cells to 2-aminobicyclo[2,2,1]heptane-2-carboxylic acid.

In insulin-producing cells of the RINm5F line, the nonmetabolized analogue of L-leucine, 2-aminobicyclo[2,2,1]heptane-2-carboxylic acid decreases O2 consumption, lowers ATP content, and inhibits insulin release despite stimulation of both NH4 production and 14CO2 output from cells prelabeled with L-[U-14C]glutamine. The metabolic and secretory effects of 2-aminobicyclo[2,2,1]heptane-2-carboxylic acid are opposed to those of D-glucose, which increases respiration, ATP content, and insulin release, while lowering NH4+ production and 14CO2 output from the prelabeled cells. D-Glucose also antagonizes the inhibitory action of 2-aminobicyclo[2,2,1]heptane-2-carboxylic acid upon both respiration and secretion. These findings suggest that, in tumor as in normal islet cells, the regulation of insulin release by exogenous nutrients depends on the availability of endogenous ATP.

Adenoma, Islet Cell↗

PubMed1987

Interference of a nonmetabolized analogue of L-leucine with lipid metabolism in tumoral pancreatic islet cells.

The nonmetabolized analogue of L-leucine, 2-aminobicyclo[2,2,1]heptane-2-carboxylic acid (BCH), was recently found to inhibit O2 uptake and insulin release from tumoral islet cells of the RINm5F line. BCH inhibited lipogenesis, stimulated lipolysis, and severely decreased the oxidation of endogenous [U-14C]palmitate in prelabelled RINm5F cells. D-Glucose exerted metabolic effects which were sometimes opposite to those caused by BCH and, within limits, protected the islet cells against the inhibitor action of BCH. Since BCH augments NH4+ production and facilitated the catabolism of 14C-labelled amino acids in the prelabelled cells, it is proposed that the unexpected inhibition of O2 uptake by BCH is mainly attributable to a decrease in the oxidation of endogenous fatty acids.

Amino Acids↗

PubMed1987

Channeling of alpha-D-glucose 6-phosphate in tumoral islet cells exposed to D-galactose.

In human erythrocytes, in which the fractional turnover rate of glucose 6-phosphate is rather low, menadione increases to almost the same relative extent the oxidation of D-[U-14C]glucose and D-[U-14C]galactose. However, in pancreatic tumoral islet cells (RINm5F line), in which the fractional turnover rate of glucose 6-phosphate is considerably higher, menadione increases the oxidation of D-[1-14C]glucose but not that of D-[1-14C]galactose. These results suggest that alpha-D-glucose 6-phosphate generated from exogenous D-galactose is channeled preferentially into the glycolytic rather than pentose phosphate pathway. Such was no more the case, however, when the RINm5F cells were exposed simultaneously to both D-glucose and D-galactose.

Adenoma, Islet Cell↗

PubMed1987

Stimulation by D-glucose of mitochondrial oxidative events in islet cells.

D-Glucose catabolism was examined in rat pancreatic islets and tumoral insulin-producing cells (RINm5F line) exposed to D-[5-3H]glucose, D-[3,4-14C]glucose or D-[6-14C]glucose. In normal islets, a rise in D-glucose concentration from 2.8 to 16.7 mM augmented (i) the transfer of NADH, presumably via the glycerol phosphate shuttle, into the mitochondria, (ii) the decarboxylation of pyruvate and (iii) the oxidation of acetyl residues, even when the flow rates were expressed relative to the total rate of D-glucose utilization. Islet cells are thus organized to favour those mitochondrial oxidative events yielding the major fraction of the energy derived from the catabolism of D-glucose. In tumoral islet cells of the RINm5F line, all three oxidative mitochondrial processes, when expressed relative to the overall rate of D-glucose utilization, occurred at a low rate and failed to increase in response to the rise in D-glucose concentration. These findings emphasize the importance of distal regulatory processes in the control of D-glucose catabolism in normal islet cells, and document the perturbation of such processes in a model of dysfunction for this fuel-sensor organ.

Acetyl Coenzyme A↗

PubMed1987

Glucose-induced changes in cytosolic ATP content in pancreatic islets.

The cytosolic and mitochondrial contents in ATP, ADP and AMP were measured in islets incubated for 45 min at increasing concentrations of D-glucose and then exposed for 20 s to digitonin. The latter treatment failed to affect the total islet ATP/ADP ratio and adenylate charge. D-Glucose caused a much greater increase in cytosolic than mitochondrial ATP/ADP ratio. In the cytosol, a sigmoidal pattern characterized the changes in ATP/ADP ratio at increasing concentrations of D-glucose. These findings are compatible with the view that cytosolic ATP participates in the coupling of metabolic to ionic events in the process of nutrient-induced insulin release.

Adenosine Diphosphate↗

PubMed1987

Effect of cytochalasin B on glucose uptake, utilization, oxidation and insulinotropic action in tumoral insulin-producing cells.

Cytochalasin B (17-3 microM) virtually abolished 3-O-methyl-D-[U-14C]glucose uptake and D-[5-3H]glucose utilization in tumoral insulin-producing cells of the RINm5F line. This coincided with a marked decrease in D-[U-14C]glucose oxidation and suppression of the stimulant action of D-glucose upon insulin release. Cytochalasin B, however, augmented basal insulin release by the tumoral cells. The RINm5F cells appeared much more sensitive than normal islet cells to cytochalasin B, as judged by the relative magnitude of inhibition in either hexose uptake or utilization. In both cell types, the inhibitory action of cytochalasin B upon glucose metabolism seemed to be competitive, being more marked at low than high glucose concentration. These results are interpreted in support of the view that a decreased efficiency of hexose transport across the plasma membrane represents an essential deficiency of the RINm5F cells.

3-O-Methylglucose↗

PubMed1987

Metabolic and secretory response of tumoral-insulin producing cells to D-fructose and D-galactose.

At variance with normal islet cells, tumoral insulin-producing cells of the RINm5F line were found to display a positive secretory response not solely to D-glucose and D-mannose, but also to D-fructose and D-galactose. All hexoses increased the ATP/ADP ratio, exerted a sparing action upon the oxidation of endogenous nutrients in cells prelabelled with either L-[U-14C]glutamine or [U-14C]palmitate, increased the output of lactic acid and, as judged from data collected in the presence of D-[U-14C]hexoses, underwent oxidation in the RINm5F cells. The secretory response to these four hexoses appeared commensurate with the extent of their metabolic effects.

Adenine Nucleotides↗

PubMed1987

Glycerol phosphorylation and oxidation in pancreatic islets.

Homogenates of rat pancreatic islets and tumoral islet cells (RINm5F line) were found to display glycerokinase activity. In the islets like in the liver, about one-sixth of the enzyme appears bound to mitochondria. The enzymatic activities in liver and islets differ from one another, however, by their response to increasing concentrations of either glycerol or ATP and sensitivity to inhibition by D-glyceraldehyde. In intact islets, [U-14C]glycerol is efficiently oxidized, albeit at a much lower rate than that found for its phosphorylation by islet homogenates. These findings are relevant to the role played by glycerol liberated from endogenous triglycerides in the basal respiration of islet cells.

Adenoma, Islet Cell↗

PubMed1987

Hexose metabolism in pancreatic islets. Absence of glucose-6-phosphatase in rat islet cells.

Homogenates of either rat or mouse pancreatic islets, pure rat B cells or insulin-producing cells of the RINm5F line catalyzed the hydrolysis of D-glucose-6-phosphate. Relative to protein content, the enzymic activity, which was mainly associated with particulate rather than soluble subcellular material, was much lower in endocrine pancreatic cells than in liver. The rat islet enzyme differed from liver glucose-6-phosphate by its lower affinity for D-glucose-6-phosphate, its lower pH optimum, its greater relative efficiency towards L-glycerol-3-phosphate as distinct from D-glucose-6-phosphate, its restricted lability during exposure to pH 5.0, its inability to act as a glucose-6-phosphate:glucose phosphotransferase, and its insensitivity to inhibition by D-glucose. It is concluded that rat islet cells are virtually devoid of true glucose-6-phosphatase activity.

Animals↗

PubMed1987

32P, 86Rb+ and 45Ca2+ handling by tumoral insulin-secreting cells (RINm5F line).

In perifused tumoral islet cells (RINm5F line), which were prelabelled with either [32P]orthophosphate, 86Rb+ or 45Ca2+, the administration of D-glucose (1.4, 2.8 or 16.7 mM) increased the efflux of 32P, decreased the outflow of 86Rb, increased slightly the efflux of 45Ca from cells perifused in the presence of Ca2+, and decreased modestly the outflow of 45Ca from cells perifused in the absence of Ca2+. D-glucose also stimulated the net uptake of 45Ca2+. When Ba2+ (2 mM) was used, in the absence of Ca2+, instead of D-glucose as an insulin secretagogue, the efflux of 32P was little affected, but the outflow of 45Ca was dramatically increased. These changes are qualitatively similar to those occurring in normal islet cells. Nevertheless, the ionic response to D-glucose appeared, as a rule, less marked in tumoral than normal islet cells. Moreover, the concentration-response relationship was shifted to a lower range of hexose concentrations in the RINm5F cells.

Animals↗