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A R Carpinelli

Publications and source records attributed to A R Carpinelli.

At least 37 records · Page 2Linked to original sources
PubMed1992

Long-term regulation of pancreatic B-cell responsiveness to D-glucose by food availability, feeding schedule, and diet composition.

The immediate metabolic, cationic, and secretory response of the insulin-producing B-cell to D-glucose is regulated, in a delayed or long-term manner, by nutritional factors such as food availability, feeding schedule, or diet composition. The B-cell keeps the memory of these nutritional manipulations so that the corresponding changes in its responsiveness to D-glucose can be documented in vitro in isolated pancreatic islets. The results of experiments conducted in starved rats, in animals exposed to an altered feeding schedule, and in rats given free access to a high-carbohydrate, high-protein, or high-lipid, as distinct from balanced, diet all suggest that a sufficient prandial hyperglycemia is essential for maintenance of an optimal metabolic and secretory behavior of the islet B-cell in response to a rise in D-glucose concentration.

Animals↗
PubMed1992

Utilization of rat and human sera to carry out incubation and perifusion of pancreatic islets.

Incubation of isolated pancreatic rat islets in a medium consisting of fresh rat or human sera and the measurement of insulin secretion after 60 min in the presence of 6.0-7.0 and 16.7 mM of glucose, were carried out. Perifusion experiments with isolated rat islets preincubated in the presence of rat or human sera and 0.2 mCi/mL of 45Ca2+ were also performed, and 45Ca2+ outflow rate and insulin secretion were analyzed. In both experiments, the usual islet responsiveness to glucose stimulation was preserved when compared with experiments in which Krebs-Henseleit (K-H) buffer was used as the medium. The results obtained demonstrate that both rat and human sera can be used as media for islets perifusion and incubation experiments, thus allowing for the effect(s) of circulating substances on insulin release to be examined.

Adult↗
PubMed1991

Insulin secretion to glucose stimulus in pancreatic islets isolated from rats fed unbalanced diets.

To verify the effect of different energetic sources on insulin secretion, just-weaned male Wistar rats were divided into four groups fed as follows: high carbohydrate (HC), high protein (HP), high lipid (HL) and balanced (C) diets during five weeks. Body weight gain and daily food intake were similar among the four groups. Insulin content of the isolated islets was not different; however, insulin release to a high glucose concentration (16.7 mM) stimulus was clearly higher in islets isolated from rats fed a balanced diet as compared to the other groups (HC, HP and HL). The rates of insulin secretion in islet perfusates from rats fed unbalanced diets were also decreased, although 45Ca2+ outflow rate (FOR) was similar in all groups. Since the rate of U-14C-glucose oxidation was decreased in islets isolated from rats fed unbalanced diets, this could be one of the mechanisms for the reduced rates of insulin release observed. Therefore, the increased supply of specific fuels provoke metabolic alterations in B-cell leading to changes in insulin secretion.

Animals↗
PubMed1991

Hexose metabolism in pancreatic islets: pyruvate carboxylase activity.

The anaplerotic hypothesis for insulin release postulates that an increased generation of malonyl-CoA, acyl residues and diacylglycerol in nutrient-stimulated pancreatic islets may couple the catabolism of nutrient secretagogues to more distal events in the secretory sequence. In the light of this hypothesis, pyruvate carboxylase activity was measured in rat pancreatic islets using two distinct radioisotopic procedures. The first procedure is based on the conversion of oxalacetate generated from pyruvate to 14C-labelled citrate in the presence of [1-14C]acetyl-CoA and citrate synthase. The second technique involves the conversion of 14C-labelled oxalacetate generated from [1-14C]pyruvate to radioactive aspartate in the presence of L-glutamate and glutamate-oxalacetate transaminase. Pyruvate carboxylase activity amounted to 10 pmol/min per islet, was restricted to mitochondria, displayed a Km for pyruvate close to 0.4 mM, and demonstrated dependency towards ATP (apparent Ka close to 0.1 mM), Mg2+ and acetyl-CoA. It is proposed that pyruvate carboxylase activity accounts for the generation of 14C-labelled amino acids other than alanine in islets exposed to D-[3,4-14C]glucose and participates to the pyruvate/citrate shuttle for the transport of acetyl-CoA out of the mitochondria in nutrient-stimulated islets.

Acetyl Coenzyme A↗
PubMed1991

Effect of the i.v. administration of non-activated lymphocytes on rat plasma insulin and glucose levels.

Although several studies have shown the effect of cytokines on islet B cell function, the role of circulating cells in the control of insulin secretion has not been investigated. The effect of lymphocyte administration on plasma glucose and insulin levels was examined in male Wistar albino rats weighing 180-200 g. The animals were anesthetized and the jugular vein was cannulated for saline or lymphocyte injection (10(6) cells in 1 ml) and blood collection after 5, 10, 15, 20, 30, 45 and 60 min. A marked increase in plasma insulin levels (180 microU/ml as compared to 56 microU/ml in the control group, at 20 min) and an unexpected increase (P < 0.05) in blood glucose levels (at 60 min only) were observed after lymphocyte administration. Similar experiments undertaken with simultaneous administration of inhibitors of arachidonic acid metabolism, indomethacin and nordihydroguairetic acid at 1 mg/kg body weight, suggest that this effect of lymphocytes is not mediated by prostaglandins or leukotrienes.

Animals↗
PubMed1990

Insulin secretion in Walker 256 tumor cachexia.

The effect of cachexia on insulin secretion was examined in adult male rats. Isolated islets of Langerhans from Walker 256 tumor-bearing rats secreted less insulin by glucose stimuli as compared with the control group; this was accompanied by significant change in 45Ca2+ outflow rate. Reduced insulin secretion to glucose stimuli in tumor-bearing rats probably led to low insulinemia (one-third). These findings indicate that reduced insulin secretion is probably an important factor for the development of cachexia in Walker 256 tumor-bearing rats.

Animals↗
PubMed1989

The effect of controlled feeding conditions on the metabolic characteristics of rats.

To evaluate the effect of feeding conditions on blood glucose, insulin and free fatty acid concentrations, rats were maintained on a 2-hr feeding/22-hr fasting (regular-fasted) schedule for 4 weeks. The animals were then subjected to glucose or insulin loads immediately prior to the usual meal time. Animals fasted for only 22 hr (single-fasted) just before the experiments, and rats having access to food ad lib were similarly loaded and tested. The results demonstrate that the regular fasting regime induces certain metabolic alterations well described in the literature for the single-fasted-period to become more pronounced, specifically, a reduction in insulin secretion and a probably increase in peripheral responsiveness to this hormone. In addition, glucose loading was more effective in lowering plasma free fatty acid concentration in rats restricted to a regular fasting scheme.

Animals↗
PubMed1989

Insulin secretion in the isolated islets of single-, regular-fasted and fed rats.

To investigate the alterations in insulin secretion induced by habituation to single daily meal, adult rats were trained on a regular-fasting scheme (2-hr feeding/22-hr fasting) for 4 weeks. Insulin secretion induced by nutrient secretagogues (D-glucose and L-leucine) and the rates of 45Ca2+ outflow and 14C-glucose oxidation were studied in isolated islets obtained from these animals and in fed and 22-hr single-fasted rats. As expected, in the 22-hr fasted group, insulin secretion was drastically decreased although not abolished while 45Ca2+ outflow and 14C-glucose oxidation rates were only partially reduced. However, the regular-fasted rats did not secrete insulin in response to the nutrient secretagogues; calcium entry was not detected although a partial reduction in 14C-glucose oxidation rate was observed. These results suggest that regular fasting induces alterations in pancreatic B-cell glycolytic pathways leading to impairment of calcium efflux and insulin secretion. Such impairment is more pronounced than that induced by a single 22-hr fast.

Animals↗
PubMed1986

Fasting-induced dissociation of cationic and secretory events in pancreatic islets.

In pancreatic islets removed from 48 h-fasted rats, as distinct from fed animals, the release of insulin evoked by D-glucose is more severely impaired than that evoked by 2-ketoisocaproate. This decreased secretory response to D-glucose contrasts with an unimpaired cationic response to the sugar in terms of the glucose-induced decrease in both 86Rb and 45Ca outflow from pre-labelled islets. Likewise, fasting only causes a modest decrease of the secondary rise in 45Ca outflow evoked by D-glucose in islets perifused at normal Ca2+ concentration. The latter decrease appears more marked, however, if the cationic response to glucose is expressed relative to that evoked by 2-ketoisocaproate in islets removed from rats in the same nutritional state. It is concluded that, in the process of nutrient-stimulated insulin release, neither the decrease in K+ conductance (inhibition of 86Rb outflow) nor the sequestration of Ca2+ by intracellular organelles and/or direct inhibition of Ca2+ outward transport (decrease in 45Ca outflow) represent the sole determinant(s) of the subsequent gating of Ca2+ channels (secondary rise in 45Ca efflux).

Animals↗
PubMed1985

Cholinergic stimulation of ion fluxes in pancreatic islets.

Cholinergic agents are known to stimulate the hydrolysis of polyphosphoinositides in pancreatic islets. The effect of carbamylcholine upon ion fluxes in the islet cells was investigated. Carbamylcholine provoked a rapid but poorly sustained increase in 45Ca and 86Rb outflow from perifused islets. Such a cationic response was observed at different glucose concentrations (zero to 16.7 mM), at three concentrations of carbamylcholine (10 microM, 100 microM and 1.0 mM), and in the absence or presence of extracellular Ca2+. It coincided with a biphasic stimulation of insulin release, both the cationic and secretory responses being abolished in the presence of atropine (10 microM). At variance with nutrient secretagogues, carbamylcholine failed to affect the net production of cyclic AMP and caused a transient decrease in 32P outflow from islets prelabelled with [32P]phosphate. It is proposed that cholinergic agents mobilize Ca2+ from intracellular sites, possibly through generation of inositol, 1,4,5-triphosphate from phosphatidylinositol 4,5-bisphosphate. The intracellular redistribution of Ca2+ does not appear sufficient, however, to account fully for the secretory response, which may also involve activation of protein kinase C by diacylglycerol.

Animals↗
PubMed1985

The coupling of metabolic to secretory events in pancreatic islets. The possible role of glutathione reductase.

The participation of glutathione reductase in the process of nutrient-stimulated insulin release was investigated in rat pancreatic islets exposed to 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU). BCNU caused a time-and dose-related, irreversible inhibition of glutathione reductase activity. This coincided with a fall in both GSH/GSSG ratio and the thiol content of the islets. Pretreatment of the islets with BCNU inhibited the oxidation of glucose and its stimulant action upon both 45Ca net uptake and insulin release. Although BCNU (up to 0.5 mM) failed to affect the oxidation of L-leucine and L-glutamine, it also caused a dose-related inhibition of insulin release evoked by the combination of these two amino acids. The latter inhibition was apparently not fully accounted for by the modest to negligible effects of BCNU upon 45Ca uptake, 45Ca efflux, 86Rb efflux and cyclic AMP production. Since BCNU failed to inhibit insulin release evoked by the association of Ba2+ and theophylline, these results support the view that glutathione reductase participates in the coupling of metabolic to secretory events in the process of nutrient-stimulated insulin release. However, the precise modality of such a participation, for example the control of intracellular Ca2+ distribution, remains to be elucidated.

Animals↗
PubMed1982

Stimulus-secretion coupling of amino acid-induced insulin release VII. The B-cell memory for L-glutamine.

In the absence of another exogenous nutrient, L-glutamine does not stimulate insulin release from rat pancreatic islets or isolated perfused pancreases. L-glutamine, however, augments insulin release evoked by L-leucine. These two amino acids could interact by providing both the substrate (L-glutamate) and an activator (L-leucine) for the reaction catalyzed by glutamate dehydrogenase. Under suitable experimental conditions, as little as 0.5 mM L-glutamine is sufficient to enhance leucine-stimulated insulin release. When the pancreases or islets are first exposed to L-glutamine and then stimulated with L-leucine, the rate of secretion is much higher than that evoked by L-leucine in tissue not first exposed to L-glutamine. The memory of a prior exposure to L-glutamine persists for at least 25 min after removal of the latter amino acid from the extracellular fluid. This memory phenomenon is not dependent on the presence of Ca2+ in the extracellular fluid during the first exposure to L-glutamine, but is suppressed when such a prior exposure is performed in the absence of extracellular K+. The memory phenomenon could be due, in part at least, to inhibition by L-glutamine of K+ conductance in the B-cell plasma membrane. Moreover, the amount of L-glutamate which accumulates in islets exposed to L-glutamine is sufficient to maintain, for a much longer period than with other nutrient secretagogues, a sustained increase in catabolic fluxes after removal of the amino acid from the extracellular fluid.

Animals↗
PubMed1981

The stimulus-secretion coupling of amino acid-induced insulin release. IV. Ionic response to L-Leucine and L-Glutamine.

L-Glutamine enhances insulin release evoked by L-leucine in isolated rat pancreatic islets. The enhancing action of L-glutamine, which is a rapid but steadily increasing and not rapidly reversible phenomenon is not attributable to any major change in either K+ or Ca2+ outflow from the islet cells. It coincides with an apparent increase in Ca2+ inflow rate and, hence, with Ca accumulation in the islets. The initial ionic response to L-leucine is not qualitatively altered by the presence of L-glutamine. In their combined capacity to stimulate 45Ca net uptake in the islets, L-glutamine can be replaced by L-asparagine but not by L-glutamate, whereas L-leucine can be replaced by L-norvaline or L-isoleucine, but not by L-valine, glycine or L-lysine. Such a specificity is identical to that characterizing the effect of these various amino acids upon insulin release. It is postulated that the release of insulin evoked by the combination of L-leucine and L-glutamine involves essentially the same remodelling of ionic fluxes as that evoked by other nutrient secretagogues with, however, an unusual time course for the functional response to L-glutamine.

Amino Acids↗
PubMed1981

Stimulus-secretion coupling of glucose-induced insulin release. Timing of early metabolic, ionic, and secretory events.

The timing of the early metabolic, ionic, and secretory responses to glucose in rat pancreatic islets was monitored by measuring, at 12 sec intervals, the concentrations of glucose, lactic, and pyruvic acids, 32P, 86Rb, 45ca, and insulin in the effluent of perifused prelabeled islets. The increase in glucose concentration from zero to 16.7 mM was complete within 133 sec. The output of organic acids increased after 24 sec of exposure to glucose and, in the case of lactic acid, fell slightly after the initial elevation. The phosphate flush was initiated only after 96 sec of exposure to glucose, whereas the decreases in 86Rb and 45Ca outflow were both detectable within 72 sec of stimulation. The secondary rise in 45Ca efflux was first seen after 157 sec of stimulation and its time course was not vastly different from that of insulin release. These data indicate that, in the secretory sequence, metabolic changes precede both the remodelling of ionic fluxes and the stimulation of insulin release. The results are compatible with the view that the secondary rise in 45Ca outflow is attributable, in part at least, to the glucose-induced decrease in K conductance (but not to the increase in phosphate outflow), with resulting membrane depolarization and gating of voltage-dependent Ca channels.

Animals↗